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Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4dddd@¸ê$ 0|¹|.Ÿÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êø” ï `ð ÿÿÿ€€€»àã33™™™™Ì`ð ÿÿÿ–––ûßSÿ™fÌ3™f`ð ÿÿÿ€€€™ÌÿÌÌÿ33̯gÿ`ð Þöñ–––ÿÿÿ�Æÿf̨`ð ÿÿÙwwwÿÿ÷3ÌÌÿPPÿ™`ð €€ÿÿÿZXÿÿ™dbmoÇÿÿÿ`ð €ÿÿÿ\ßÒ“Ì3¾y`ÿÿ™Ó¢`ð ™ÿÿÿ3fÌÿÿ3f̰fÌÿÿç`ð ÿÿÿ3f™ãëñ3™FŠKfÌÿðå`ð hk]ÿÿÿwwwÑÑË��‚€ž¨ÿÌféܹ`ð ff™ÿÿÿ>>\ÿÿÿ`Y{ffÿ™Ìÿÿÿ™`ð R>&ÿÿÿ- ßÀ�Œ{p�_/Ì´Œž £>ÿý?" dÿd ÿÿïÿÿÿÿÿÿ²£|ÿý?" dÿdi ÿÿïÿÿÿÿÿÿ�€ cŒq€" ]  a€ é£ Q€»u/ £nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ   @@``€€P£R  Œ   £  /`£ p£>qaQII€£>aQIAA Æð¾ððVð( ð ððð ð ã ðT€„ƒŸ�\£‚®Ñƒ\£„®Ñ‡¿�ƒ¿Àÿ ðÄ€ƒUIðà Ÿ ðTŸ¨ Click to edit Master title style¢!ª !ð4 ð Ó ðN€†Ÿ�\£‚®Ñƒ\£„®Ñ¿�ƒ¿Àÿ 𠀃Uº=ðà Ÿ 𞟨RClick to edit Master text styles Second level Third level Fourth level Fifth level¢!    ª Sðò ð Ó ðN€,“Ÿ�\£‚®Ñƒ\£„®Ñ¿�ƒ¿Àÿ ðí>€�ºCðà Ÿ ð\Ÿ *¡9ø¦ñ ŒŒ  £ £ //ðô ð Ó ðN€t–Ÿ�\£‚®Ñƒ\£„®Ñ¿�ƒ¿Àÿ ðí>ÁB;ºCðà  Ÿ ð^Ÿ *¡9ú¦ñ ŒŒ  £ £ //ðô ð Ó ðN€Ä Ÿ�\£‚®Ñƒ\£„®Ñ¿�ƒ¿Àÿ ðí>‚@ƒUºCðà Ÿ ð^Ÿ *¡9ئñ ŒŒ  £ £ //ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ8Š0º___PPT10‹ë.+OÅZ³â ºDefault DesignîWï € Çð¿ ð ðWð( ð ðð¢ ð Ó ð®€ä�5�\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�ƒ¿ Àÿ?¿ÿ?„†ˆ#"ñ Œ�0eðó^!=* ðà  5 ð ž悁ð £ ð<€’5…‡¿�ƒ¿Àÿð{¹>/Er ðCŸ¨photo¡I¦ ð³¢ ð £ ð<€Ü¡5…‡¿�ƒ¿Àÿð›[I›U’  ðGŸ¨ MOAC logo¡ I¦ ð ¢ ð ƒ ð0€„£5¿�ƒ¿Àÿ𺠋PØ ð@Ÿ ¼Fundamental to a cell is its plasma membrane: it protects the contents, provides controlled interaction with the outside, and& Design of plastic vesicles prompted by need to control the characteristics of the membrane for efficient drug delivery, robustness blah blah.. Various diblock copolymers used, as like lipids: amphiphilic and so form vesicles spontaneously when water added to them in solution Rafts in plasma membrane support proteins vital to cell function& 2 species¡ßß,ª> " •¦ ðw¢ ð £ ð<€¼5…‡¿�ƒ¿Àÿð+8¹%£@ 𠟨�Acknowledgements Supervisor: Dr Stefan AF Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Funding: EPSRC¡‚‚,ª>F ¦ ð´¢ ð £ ð<€¸Å5…‡¿�ƒ¿ÀÿðÖ7QLšXÍ: ðHŸ¨ references¡ I¦ ðµ¢ ð £ ð<€”Ï5…‡¿�ƒ¿Àÿð(öMÎY+ ðIŸ¨ Lipid rafts¡ I¦ ð¹¢ ð £ ð<€pÙ5…‡¿�ƒ¿Àÿð/jFqYø1 ðMŸ¨Plasma membrane¡I¦ ðÒ¢ ð £ ð<€ðÚ5…‡¿�ƒ¿Àÿð€æ%1Î2 ðfŸ ²block-copolymers of amphiphilic nature can assemble into different structures, such as micelles, rods, 3-D  gel-type networks, and vesicles. How to make polymersomes Dissolve plastic in tetrahydrofuran (THF) which is a good solvent and subsequently altering the composition of the medium to different ratios of THF:water via slow addition of water (a bad solvent). Samples were taken at solvent compositions 10-80% water and analysed by Transmission Electron Microscopy (TEM). Negative staining techniques using a 1% solution of Uranyl Acetate were used to enhance the contrast of the micrographs.¡ Z,U,ªb } o Ñ?¦ ðT¢ ð £ ð<€ðì5…‡¿�ƒ¿Àÿð{¼MHqò' ð蟠 Polymer 1 (2 g/L)  80% water Once water is the major solvent rods dominate the sample and donuts appear with sizes up to 400 nm. Scale bar equals 200 nm. Polymer 2 (5 g/L)  50% water Larger spheres become more pronounced. A very regular pattern appears. The larger spheres could be vesicles. Scale bar equals 200 nm. Polymer (5 g/L) 3  50% water Under these conditions only vesicles can be see. The observed bilayer thickness is 20 nm. Scale bar equals 500 nm ¡ÑÐ(I¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ8Š0º___PPT10‹ë.+OÅ à´âî*ï € )ð)0ð!8ñð�(ð( ð ðð( ð 3 ð‚€€€„ÚŸ…‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€��ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€ 5¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€ì5�€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð¢ ð  ƒ ð0€”5¿�ƒ¿ÀÿðòbAX" 𷟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡,  ($ª>V hH¦ 𻢠ð ó ðj€p05¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€œI5¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€d25�\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  5 ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ðÖ¢ ð/ ó ðj€@i5¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðì2ñ9W¦: ðþŸ lFurther work I am glad to report that this has been a useful contribution to Stefan Bon s ressearch group coninues research continues in order to reconcile the finidngs and classify¡"· (ª(ª>Z /¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€dt5¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà 5ð´¢ ð2 ³ ðB€˜’N ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËWuA ðŸ¨XReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N M  $   ª,6¦ ðð8 ð1N# j7 ð6ðº"ÖÅÖ6ðXð@ ð1�% j7 ð(ð1�% j7ð¾ð@ ð1�% ö4 ð%ð1�% ö4ð²² ð c ðt€€AÁP¿ÿpolymerzomes with rafts_75000X_25 small"ñ¿`ð1�%AÉ4ð´² ð c ðv€€AÁR¿ÿpolymerzomes with rafts_300000X_23 small"ñ¿`ð½% ö4ðB¢ ð&  ðv€ÄR5…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð °5 j7 ðVŸ¨Negative stain TEM¡(™þ¦ ðf¢ ð3  ðv€;N …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðvN#0% ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðð8 ð#Z/ 9ÞA ð7ð0"! 7ˆBðZð@ ðñ9s&WH7 ð+ðV'B1¼5ÞAð°² ð c ðr€€AÁN¿ÿ 20% Brij polymersomes-32250X-115 small"ñ¿`ðñ9s&WHÙ4ðR¢ ð*  ðv€X]5…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð>U5fD7 ðfŸ¨Cryo-TEM¡  (fþª¦ ðe¢ ð4  ðv€ŒèN …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð#Z/ 91 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ð²ð8 ðŸC ¥VR2 ð8ðzŸC¥V®0ðð@ ðaHË#Z„3 ð-ð¨D™"JVR2ðz² ð c ð<€€AÁ¿ÿrichard1-81"ñ¿`ðaHË#Z1ð3¢ ð,  ðv€ _5…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðéOÊ1S„3 ðGŸ¨SEM¡(ffþ¦ ð]¢ ð5  ðv€,óN …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ 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ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0¸ÿÿÿüpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîÑ)ï € è(ðà(0ð9@ñð`(ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€èyá ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðòbAX" 𥟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡,  ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢   !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~€�‚ƒ„…†‡ˆ‰Š‹Œ�Ž��‘’“”•–—˜™š›œ�žŸ ¡¢£¤¥¦§¨©ª«¬­®¯°±²³´µ¶·¸¹º»¼½¾¿ÀÁÂÃÄÅÆÇÈÉÊËÌÍÎÏÐÑÒÓÔÕÖרÙÚÛÜÝÞßàáâãäåæçèéêëìíîïðñòóôõö÷øùúûüýþÿ      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~€�‚ƒ„…†‡ˆ‰Š‹Œ�Ž��‘’“”•–—˜™š›œ�žŸ ¡¢£¤¥¦§¨©ª«¬­®¯°±²³´µ¶·¸¹º»¼½¾¿ÀÁÂÃÄÅÆÇÈÉÊËÌÍÎÏÐÑÒÓÔÕÖרÙÚÛÜÝÞßàáâãäåæçèéêëìíîïðñòóôõö÷øùúûüýþÿ      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~€�‚ƒ„…†‡ˆ‰Š‹Œ�Ž��‘’“”•–—˜™š›œ�žŸ ¡¢£¤¥¦§¨©ª«¬­®¯°±²³´µ¶·¸¹º»¼½¾¿ÀÁÂÃÄÅÆÇÈÉÊËÌÍÎÏÐÑÒÓÔÕÖרÙÚÛÜÝÞßàáâãäåæçèéêëìíîïðñòóôõö÷øùúûüýþÿ      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~€�‚ƒ„…†‡ˆ‰Š‹Œ�Ž��‘’“”•–—˜™š›œ�žŸ ¡¢£¤¥¦§¨©ª«¬­®¯°±²³´µ¶·¸¹º»¼½¾¿ÀÁÂÃÄÅÆÇÈÉÊËÌÍÎÏÐÑÒÓÔÕÖרÙÚÛÜÝÞßàáâãäåæçèéêëìíîïðñòóôõö÷øùúûüýþÿ      !"#$%&'þÿÿÿ)*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTU‘WXYZ[\]^_`abcdefghijklmnopqrst®vwxyz{|}~€�‚ƒ„²ýÿÿÿýÿÿÿýÿÿÿýÿÿÿýÿÿÿýÿÿÿýÿÿÿýÿÿÿýÿÿÿýÿÿÿ�þÿÿÿ’“”•–—˜™š›œ�žŸ ¡¢£¤¥¦§¨©ª«¬­uÒþÿÿÿ±þÿÿÿ³´µ¶·¸¹º»¼½¾¿ÀÁÂÃÄÅÆÇÈÉÊËÌÍÎÏÐÑÒÓÔÕÖרÙÚÛÜÝÞßàáâãäåæçèéêëìíîïðñòóôõö÷øùúûüýþýÿÿÿRoot Entryÿÿÿÿÿÿÿÿ��d›Oφꪹ)è0žKk{[ŰÀPicturesÿÿÿÿÿÿÿÿ;NCurrent Userÿÿÿÿÿÿÿÿÿÿÿÿ JSummaryInformation(ÿÿÿÿV€PPowerPoint Document(ÿÿÿÿÿÿÿÿÿÿÿÿ(^vDocumentSummaryInformation8ÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿ @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 82 9 ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹�§=ÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîò)ï €  )ð)0ð!8ñð�(ð( ð ðð( ð 3 ð‚€€€|P …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€¤´ ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€Xå=¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€P �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð¢ ð  ƒ ð0€,{ ¿�ƒ¿ÀÿðòbAX" 𥟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡,  ($ª,V hO¦ 𻢠ð ó ðj€è©,¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ÈI¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€$I�\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  I ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ðØ¢ ð/ ó ðj€` I¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðì2ñ9W¦: ðŸ lFurther work I am glad to report that this has been a useful contribution to Stefan Bon s ressearch group coninues research continues in order to reconcile the finidngs and classify¡$· (ª(ª>Z /¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€|.I¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà Ið´¢ ð2 ³ ðB€x?I¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËWuA ðŸ¨XReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N M  $   ª,6¦ ðð8 ð1N# j7 ð6ðº"ÖÅÖ6ðXð@ ð1�% j7 ð(ð1�% j7ð¾ð@ ð1�% ö4 ð%ð1�% ö4ð²² ð c ðt€€AÁP¿ÿpolymerzomes with rafts_75000X_25 small"ñ¿`ð1�%AÉ4ð´² ð c ðv€€AÁR¿ÿpolymerzomes with rafts_300000X_23 small"ñ¿`ð½% ö4ðB¢ ð&  ðv€`BI…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð °5 j7 ðVŸ¨Negative stain TEM¡(™þ¦ ðf¢ ð3  ðv€p_I…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðvN#0% ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðð8 ð#Z/ 9ÞA ð7ð0"! 7ˆBðZð@ ðñ9s&WH7 ð+ðV'B1¼5ÞAð°² ð c ðr€€AÁN¿ÿ 20% Brij polymersomes-32250X-115 small"ñ¿`ðñ9s&WHÙ4ðR¢ ð*  ðv€lI…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð>U5fD7 ðfŸ¨Cryo-TEM¡  (fþª¦ ðe¢ ð4  ðv€üsI…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð#Z/ 91 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ð²ð8 ðŸC ¥VR2 ð8ðzŸC¥V®0ðð@ ðaHË#Z„3 ð-ð¨D™"JVR2ðz² ð c ð<€€AÁ¿ÿrichard1-81"ñ¿`ðaHË#Z1ð3¢ ð,  ðv€pwI…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðéOÊ1S„3 ðGŸ¨SEM¡(ffþ¦ ð]¢ ð5  ðv€l‰I…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðŸC ¥VØ! ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rÖZgjõm²Za”Gè‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿÿÿÿÿÞÝÞâÂâÝÝÿÿÿÿÿÿÿÿÿùùÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿâÿÞÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÃÿÿÃÂÃâÂÃâÃâââÃÿâÃÃâÃâÿÂÂÿÃÃââÂÃÃÃâÂÿÃÃÃÃâÂÿÃÂÿÿÿÿÿÿÿÿÿÿÿÿÿùùÿÿÿÿÿÿÿÝÂÝÝâÝÿÂâÿÿÿÿÿÿÿÿÿùùÿÿÃÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿâÞâÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÂâÂÃâÃÃâÿâÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿùùùùÿÿÿ½ÂÂâÝÿÿÿÿÿÿÿÿÿÿùùùÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÃÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÃâÃÿÃÂÿÃââÃââÃâÿâÃâââÃâÃÃâÃÂâÃÃÿÿÿÂÃÿâÃÂÿÃâÃÿÂÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿùùùùùùùÿÿÿÿùùùùùùùÿÿÿÿÿÿÿÿÿÿþÿÿÿ þÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿþÿÕÍÕœ.“—+,ù®0؈� ¬´¼Ä Ì ÔÜ äìôü  xäCustomuow^v%{ Comic Sans MSTimes New RomanDefault Design/Seeking rafts and finding holes Richard Wilson  Fonts UsedDesign Template Slide Titlesmplate Slide Titlesö&_À‘ã:vôŸRichard WilsonRichard Wilson ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ðØ¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðì2ñ9W¦: ðŸ lFurther work I am glad to report that this has been a useful contribution to Stefan Bon s ressearch group coninues research continues in order to reconcile the finidngs and classify¡$· (ª(ª>Z /¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ð´¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËWuA ðŸ¨XReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N M  $   ª,6¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ð*ð8 ð#Z/ 9ÞA ð7ð0"! 7ˆBðuð@ ð÷8s&uI7 ð+ð\&B1Ú6ÞAð°² ð c ðr€€AÁN¿ÿ 20% Brij polymersomes-32250X-115 small"ñ¿`ðñ9s&WHÙ4ðm¢ ð*  ðv€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð÷8U5uI7 ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðe¢ ð4  ðv€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð#Z/ 91 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðÍð8 ðŸC ¥VR2 ð8ðzŸC¥V®0ð ð@ ðaHË#Z„3 ð-ð¨D™"JVR2ðz² ð c ð<€€AÁ¿ÿrichard1-81"ñ¿`ðaHË#Z1ðN¢ ð,  ðv€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðÚJÊ1-X„3 ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð]¢ ð5  ðv€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðŸC ¥VØ! ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +r�”.¤õmy”Îl è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 80 : ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0øüpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîÝ(ï € ô'ðì'0ð9@ñðl'ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€èyá ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðòbAX" 𥟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡,  ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ðä¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðì2ñ9W¦: 𠟨žConclusion Mixing two different polymers has resulted in unusual structures that will probably be useful but to produce rafts requires a different combination¡"Ÿ (”(ª“ ¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ð´¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËWuA ðŸ¨XReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N M  $   ª,6¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ð*ð8 ð#Z/ 9ÞA ð7ð0"! 7ˆBðuð@ ð÷8s&uI7 ð+ð\&B1Ú6ÞAð°² ð c ðr€€AÁN¿ÿ 20% Brij polymersomes-32250X-115 small"ñ¿`ðñ9s&WHÙ4ðm¢ ð*  ðv€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð÷8U5uI7 ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðe¢ ð4  ðv€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð#Z/ 91 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðÍð8 ðŸC ¥VR2 ð8ðzŸC¥V®0ð ð@ ðaHË#Z„3 ð-ð¨D™"JVR2ðz² ð c ð<€€AÁ¿ÿrichard1-81"ñ¿`ðaHË#Z1ðN¢ ð,  ðv€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðÚJÊ1-X„3 ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð]¢ ð5  ðv€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðŸC ¥VØ! ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rCÎÔÝõmιl è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 80 < ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0øbpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êî )ï € 7(ð/(0ð;`ñð§'ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€èyá ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðòbAX" 𥟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡,  ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð ¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð20b:ˆW®8 ð4Ÿ¨èConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts requires a different combination of polymers.¡"é (Þ$¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ðµ¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËW¨B ðŸ¨YReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N N  $   ª,7¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ð*ð8 ð#Z/ 9ÞA ð7ð0"! 7ˆBðuð@ ð÷8s&uI7 ð+ð\&B1Ú6ÞAð°² ð c ðr€€AÁN¿ÿ 20% Brij polymersomes-32250X-115 small"ñ¿`ðñ9s&WHÙ4ðm¢ ð*  ðv€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð÷8U5uI7 ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðe¢ ð4  ðv€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð#Z/ 91 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðírCxV«-ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rõ†õmÑ®?l è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0øüpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîê(ï € (ðù'0ð<€ñ ði'ð( ð      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}ýÿÿÿ€ ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€èyá ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðòbAX" 𥟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡,  ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð ¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð20b:ˆW®8 ð4Ÿ¨èConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts requires a different combination of polymers.¡"é (Þ$¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ðµ¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËW¨B ðŸ¨YReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N N  $   ª,7¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðírCxV«-ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rê?{OõmÆ?mxl è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîü*ï € *ð *0ð<€ñ ð{)ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðÆ¢ ð ƒ ð0€èyá ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔŒ ðXŸ¨¼Why seek rafts? Rafts are areas of different species of lipid present in the plasma membrane of a cell. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,­(­$ª>  #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ðû¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðòbAX" 𛟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡" ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð20b:ˆWb; ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ðµ¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËW¨B ðŸ¨YReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N N  $   ª,7¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðírCxV«-ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +r©x:ˆõm…x>³l è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êî++ï € B*ð:*0ð<€ñ ðª)ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðõ¢ ð ƒ ð0€èyá ¿�ÿ‚33¿Àÿ"ñ¿`ðNÔæ ð‡Ÿ¨ÙWhy seek rafts? Animal cell plasma membranes have areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ê(Ê$ªP8€ #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ðû¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðòbAX" 𛟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡" ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð"¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðJŸ ÀA polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed out of a phospholipid bilayer membrane.¡,á$ $Ô$ª, µ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð20b:ˆWb; ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ðµ¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËW¨B ðŸ¨YReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 ¡N N  $   ª,7¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðírCxV«-ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rz³ ÃõmV³>îl è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîÝ+ï € ô*ðì*0ð<€ñ ð\*ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðå¢ ð ƒ ð0€èyá ¿�ÿ‚š¿Àÿ"ñ¿`ðNÔŒ ðwŸ¨ÛWhy seek rafts? Animal cell plasma membranes include areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ì(Ì$ª>¿ #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ðû¢ ð  ƒ ð0€T¤á ¿�ƒ¿Àÿðò4AâW" 𛟨Copolymers The plastics used in these experiments were PEO-PMMA and Brij-97. Both are amphiphilic copolymers: formed from two different polymers, one hydrophilic, one hydrophobic, covalently combined. Brij is much shorter than PEO-PMMA and has a double bond in its hydrophobic chain.¡" ($ª,V hO¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðFŸ ¼A polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed from a phospholipid bilayer membrane.¡,ß$ $Ò$ª, ³ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðª/�:µWÚ: ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÖð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bð¬¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð*Ÿ `Acknowledgements Supervisor: Dr Ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6    ªb!( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ð{¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðk<–9ËWÛC ðËŸ¨ßReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 Wolfgang H Binder, Veronique Barragan and Frederic M Menger, Domains and rafts in Lipid Membranes, Agew. Chem. Int. Ed., 2003, 42, 5802¡X Õ  $  V $ ' ªbR(!¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðírCxV«-ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rzî þõmVîð)Ý è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êî<-ï € S,ðK,0ð<€ñ ð»+ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðå¢ ð ƒ ð0€èyá ¿�ÿ‚š¿Àÿ"ñ¿`ðNÔŒ ðwŸ¨ÛWhy seek rafts? Animal cell plasma membranes include areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ì(Ì$ª>¿ #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð\¢ ð  ƒ ð0€T¤á ¿�ƒ¿Àÿð=R@WÇ ðüŸ >Copolymers The plastics used in these experiments were PEO45-PMMA180 (polyethylene oxide-polymethyl methacrylate) and Brij-97 (polyoxyethylene-10-oleyl-ether). Both are copolymers - formed by covalently bonding two different polymers  and amphiphilic - one hydrophilic, one hydrophobic.¡N  (/$ $çÿ$ $çÿÜ$ª>Y  € %¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðFŸ ¼A polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed f�‚ƒ„…†‡ˆ‰Š‹Œ�Ž��‘’“”•–—˜™š›œ�žŸ ¡¢£¤¥¦§¨©ª«¬­®¯°±²³´µ¶·¸¹º»¼½¾¿ÀÁÂÃÄÅÆÇÈÉÊËÌÍÎÏÐÑÞÓÔÕÖרÙÚþÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿßàáâãäåæçèéêëìíîïðñòóôõö÷øùúûýýÿÿÿþÿrom a phospholipid bilayer membrane.¡,ß$ $Ò$ª, ³ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðª/�:µWÚ: ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÔð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bðª¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð(Ÿ ^Acknowledgements Supervisor: Dr ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6Ÿ Ÿ ªb ( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ð{¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nðã;­9âWSC ðËŸ¨ßReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 Wolfgang H Binder, Veronique Barragan and Frederic M Menger, Domains and rafts in Lipid Membranes, Agew. Chem. Int. Ed., 2003, 42, 5802¡X Õ  $  V $ ' ªbR(!¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðÛrCxV™.ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +r,*½9õm*gÝ è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0øüpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êî<-ï € S,ðK,0ð<€ñ ð»+ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðå¢ ð ƒ ð0€èyá ¿�ÿ‚š¿Àÿ"ñ¿`ðNÔŒ ðwŸ¨ÛWhy seek rafts? Animal cell plasma membranes include areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ì(Ì$ª>¿ #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð\¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðâR@Wl ðüŸ >Copolymers The plastics used in these experiments were PEO45-PMMA180 (polyethylene oxide-polymethyl methacrylate) and Brij-97 (polyoxyethylene-10-oleyl-ether). Both are copolymers - formed by covalently bonding two different polymers  and amphiphilic - one hydrophilic, one hydrophobic.¡N  (/$ $çÿ$ $çÿÜ$ª>Y  € %¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ð¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðFŸ ¼A polymersome is an artificial vesicle with a bilayer membrane composed of plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed from a phospholipid bilayer membrane.¡,ß$ $Ò$ª, ³ ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðô.¼:âW$: ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÔð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bðª¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð(Ÿ ^Acknowledgements Supervisor: Dr ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6Ÿ Ÿ ªb ( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ð{¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðÀ;Ô:kXrB ðËŸ¨ßReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 Wolfgang H Binder, Veronique Barragan and Frederic M Menger, Domains and rafts in Lipid Membranes, Agew. Chem. Int. Ed., 2003, 42, 5802¡X Õ  $V$'ªbR(!¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðârCxV -ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +r=gÎvõmg¤Ý è‰é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MSÌæ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8/ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð­�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0�ýÿÿRÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹¤æÿ ˆ²Š0º___PPT10‹ pµpµŠrº___PPT9‹T®L¯¬¯¬?Ù Ú %ðjóŸ NSeeking plastic rafts& Richard Wilson ¡0(²(" 26,Ÿª óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîf-ï € },ðu,0ð<€ñ ðå+ð( ð ðð( ð 3 ð‚€€€Hgá …‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðå¢ ð ƒ ð0€èyá ¿�ÿ‚š¿Àÿ"ñ¿`ðNÔŒ ðwŸ¨ÛWhy seek rafts? Animal cell plasma membranes include areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ì(Ì$ª>¿ #› 5¦ ðâ¢ ð  ƒ ð0€Ø˜á ¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€\�á �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð\¢ ð  ƒ ð0€T¤á ¿�ƒ¿ÀÿðâR@Wl ðüŸ >Copolymers The plastics used in these experiments were PEO45-PMMA180 (polyethylene oxide-polymethyl methacrylate) and Brij-97 (polyoxyethylene-10-oleyl-ether). Both are copolymers - formed by covalently bonding two different polymers  and amphiphilic - one hydrophilic, one hydrophobic.¡N  (/$ $çÿ$ $çÿÜ$ª>Y  € %¦ 𻢠ð ó ðj€˜ªá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ðH¢ ð ó ðj€ºá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðpŸ ÔA polymersome is an artificial vesicle with a bilayer membrane composed of amphiphilic plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed from a phospholipid bilayer membrane.¡,ë$ $Þ$ª> > v ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€\¶á �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  á  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ŒÔá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðô.¼:âW$: ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÔð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bðª¢ ð £ ð<€8âá ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð(Ÿ ^Acknowledgements Supervisor: Dr ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6Ÿ Ÿ ªb ( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà á ð{¢ ð2 ³ ðB€tä ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðÀ;y:XrB ðËŸ¨ßReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 Wolfgang H Binder, Veronique Barragan and Frederic M Menger, Domains and rafts in Lipid Membranes, Agew. Chem. Int. Ed., 2003, 42, 5802¡X Õ  $V$'ªbR(!¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Ðñá ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€| ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€pä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€*ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðârCxV -ðT¢ ð, # ð|€P-ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€Ø>ä …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rN¤ß³õm*¤MáÝ è�é(ZEà€ òd/È 0ÒÕ˜·DComic Sans MS$g¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ(¥ˆ. ©  @£nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8$ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   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ð€ 8$ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð‡�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0þþÿÿjÿÿÿpû�"pû-<ý4ddddL¸ê$ 0ˆ¹´šÿ ˆŒŠ0º___PPT10‹ pµpµŠLº___PPT9‹.®&¯¬?Ù Ú %ð–óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 H<êîˆ-ï € Ÿ,ð—,0ð<€ñ ð,ð( ð ðð( ð 3 ð‚€€€,…‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðˆêOE ð8Ÿ¡(¦ ðå¢ ð ƒ ð0€Œ€� ¿�ÿ‚š¿Àÿ"ñ¿`ðNÔŒ ðwŸ¨ÛWhy seek rafts? Animal cell plasma membranes include areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ì(Ì$ª>¿ #› 5¦ ðâ¢ ð  ƒ ð0€¨¿�ƒ¿Àÿð«1w5 ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+!(+$ªt  #Kh ¦ù  3T|�Åæðþ2 ð  C ðˆ€„€�¢� �€ü ‚€ü ƒ€ü „€ü …‡ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ðÅÿ&Aì4 ð8Ÿ¡-¦ ð~¢ ð  ƒ ð0€ü‰� ¿�ƒ¿ÀÿðâR@Wl ðŸ `Copolymers The plastics used in these experiments were PEO45-PMMA180 (polyethylene oxide-polymethyl methacrylate) and Brij-97 (polyoxyethylene-10-oleyl-ether). Both are copolymers - formed by covalently bonding two different polymers  and amphiphilic - one polymer is hydrophilic, the other hydrophobic.¡N1 (/$ $çÿ$ $çÿí$ª>Y  € 6¦ 𻢠ð ó ðj€ì¢� ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð]×=×/ ð㟨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j0N,,™þ", ,fþD,,ffþ,ª, Ñ !¦ ðH¢ ð ó ðj€p®� ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðå @ ZW- ðpŸ ÔA polymersome is an artificial vesicle with a bilayer membrane composed of amphiphilic plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed from a phospholipid bilayer membrane.¡,ë$ $Þ$ª> > v ¦ ð ð8 ð'{®?« ð0ðó@ ÇW#ð<¢ ð"  ðÆ€¸ó� �\£‚®Ñƒ\£„®Ñ…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  �  ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ð(¢ ð/ ó ðj€ ±� ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðô.¼:âW$: ðPŸ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r (g$ª-=¦ ðÔð8 ð1Ã9ÿfB ð1ðú9Ö¤�Bðª¢ ð £ ð<€èÝ� ¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËfB ð(Ÿ ^Acknowledgements Supervisor: Dr ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6Ÿ Ÿ ªb ( 1¦ ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà � ð{¢ ð2 ³ ðB€˜î� ¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?NðÀ;y:XrB ðËŸ¨ßReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 Wolfgang H Binder, Veronique Barragan and Frederic M Menger, Domains and rafts in Lipid Membranes, Agew. Chem. Int. Ed., 2003, 42, 5802¡X Õ  $V$'ªbR(!¦ ð«ð8 ðÖõ Ź5 ð9ðü!ÖÅÀ6𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ðm¢ ð&  ðp€Äñ� ¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 ¹5 ð‡Ÿ¨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'((™þ¦ ðl¢ ð3 # ð|€p� …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðõ Õ¯" ðzŸ¨Result 1: a weird structure!¡4(™þ(™þ(¦ ðìð8 ð«!0•7ÞA ð<ðò0«!•7ÌBð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@ðs¢ ð* # ð|€àr� …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð_$$@Ý4ÞA ð�Ÿ¨Cryo-TEM scale bar 100 nm¡$ (fþ(ª¦ ðk¢ ð4 # ð|€Ü5� …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð«!0•7¾1 ðyŸ¨%Result 2: an aggregation of vesicles?¡*&(fþ(fþ¦ ðßð8 ðrCíxV«- ð;ðârCxV -ðT¢ ð, # ð|€œA� …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð…Fñ+ØS«- ðbŸ¨SEM scale bar 100 nm¡$(ffþ(¦ ð;ð@ ðrCíxV, ð:ðrCíxV,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ðc¢ ð5 # ð|€<� …‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðrCíxV§ ðqŸ¨Result 3: perforated membrane¡*(ffþ(ffþ¦ ðH ð ƒ ð0�ƒ“iF.”‰›¬¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +rTðëþõm0ð{,‰ è�é(‚J 4à€ òd/È 0ÒÕ˜·DComic Sans MS¤.Ÿ¸ð·¡Ð0¸:A 0¸B·DTimes New Roman¸ð·¡Ð0¸:A 0¸¤ €Bÿÿ ¥ˆ.D©  @£nÿý?" dÿ      !þÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿÿd@ÿÿïÿÿÿÿÿÿ  @@``€€ ~ ðv ð€ 8$ = ŸðŒRð$àDEºò›66òî±e¡ÿ .Rð$ÜV �wu�£ö³¨áÿD- .ð$ÿð$ÿRð$þZ‘�ŽP1ÙZåÿƒPÿÕ Q[Rð$1c�~ðåÆÑ|aO¨âãmÿ( &fRð$R#o€è0ŒbŸy£ Ð9ÿL-Nsbð$r`kåKd)hÿæ8U ð�0e‚˜²ƒ0e„˜²…‡ˆ‰¿ ô   €�ÿÿÿ‚ƒÿÿÿ„… †A‡Áˆ‰Š‹Œ�Ž��‘’“”•–—Á˜™š›œ@¿ÀÁÂÿÿÿà ÄÅAÆÁÇÈÉÊË øÌÍÎÏÁ×ÿËËË 8c8c     ?€�‚ƒœ1„…ðù†‡÷ˆ ¿ÀÁÂdÃÄÅÆÇÈÉÊ0uËÐÌ0íìÿÍ@T‰Î€Ï€ÿÿÐyÿÑ2Ò NÓPÃÔÕ'Öp”×°<ÿÿØÙ'Úp”ÿA¨)BCD|¾E„|¾…†|¾‡vesicles "ñ Œ�0e@žÿÿÿÿŸÿÿÿÿ  ¡Á¢ÿÿÿÿ£ÿÿÿÿ¤ ¥Á¦ÿÿÿÿ§ÿÿÿÿ¿ ÙÿÿÿÿÚÿÿÿÿÛ ÜÁÝÿÿÿÿÞÿÿÿÿß àÁáÿÿÿÿâÿÿÿÿÿÀÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ Âÿÿÿÿÿÿÿÿ‰ÿÿÿÿŠÿÿÿÿ‹ ŒÂ�ÿÿÿÿ��‘’¿‚‚@ABÿÿÿC DEEFÅGHIJK5%LMNOÅPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Å]ÿÿÿÿ^ÿÿÿÿ_ `ÅaÿÿÿÿbÿÿÿÿN€�‚ÿÿÿƒ „…E†Å‡ˆ‰Š‹5%Œ�Ž�Å�‘’“”•—™ÿÿÿÿšÿÿÿÿ› œÅ�ÿÿÿÿžÿÿÿÿŸ  Å¡ÿÿÿÿ¢ÿÿÿÿ¿NÀÁÂÿÿÿà ÄÅEÆÅÇÈÉÊË5%ÌÍÎÏÅÐÑÒÓÔÕ×ÙÿÿÿÿÚÿÿÿÿÛ ÜÅÝÿÿÿÿÞÿÿÿÿß àÅáÿÿÿÿâÿÿÿÿÿNÿÿÿ FÆ   5%  Æÿÿÿÿÿÿÿÿ Æÿÿÿÿÿÿÿÿ Æ!ÿÿÿÿ"ÿÿÿÿ?N@ABÿÿÿC DEFFÆGHIJK5%LMNOÆPQRSTUWYÿÿÿÿZÿÿÿÿ[ \Æ]ÿÿÿÿ^ÿÿÿÿ_ `Æaÿÿÿÿbÿÿÿÿ@ñÿ™fff@ñÿ÷ðó€Ð‡�/P Êš;¨ n8Êš;úgþý4dd ¸:A 0bþÿÿ‚ÿÿÿpûPpûA%<ý4ddddL¸ê$ 0ˆ¹|.Ÿÿ ˆŒŠ0º___PPT10‹ pµpµŠLº___PPT9‹.®&¯¬?Ù Ú %ð–óŸ¨.Seeking rafts and finding holes Richard Wilson¡0/²(" 2 :0êø” ï `ð ÿÿÿ€€€»àã33™™™™Ì`ð ÿÿÿ–––ûßSÿ™fÌ3™f`ð ÿÿÿ€€€™ÌÿÌÌÿ33̯gÿ`ð Þöñ–––ÿÿÿ�Æÿf̨`ð ÿÿÙwwwÿÿ÷3ÌÌÿPPÿ™`ð €€ÿÿÿZXÿÿ™dbmoÇÿÿÿ`ð €ÿÿÿ\ßÒ“Ì3¾y`ÿÿ™Ó¢`ð ™ÿÿÿ3fÌÿÿ3f̰fÌÿÿç`ð ÿÿÿ3f™ãëñ3™FŠKfÌÿðå`ð hk]ÿÿÿwwwÑÑË��‚€ž¨ÿÌféܹ`ð ff™ÿÿÿ>>\ÿÿÿ`Y{ffÿ™Ìÿÿÿ™`ð R>&ÿÿÿ- ßÀ�Œ{p�_/Ì´Œž £>ÿý?" dÿdDÿÿïÿÿÿÿÿÿŽ£|ÿý?" dÿdºDÿÿïÿÿÿÿÿÿg€ ç¢Z€"  DN€ ¶ å A€»X‡ £nÿý?" dÿd@ÿÿïÿÿÿÿÿÿ   @@``€€P£R  ¢ D å  ‡`£ p£>ZNA::€£>NA:44 Æð¾ððVð( ð ððð ð ã ðT€œƒŸ�?�‚Ÿ@ƒ?�„Ÿ@‡¿�ƒ¿Àÿ ðºÈFá ðà Ÿ ðTŸ¨ Click to edit Master title style¢!ª !ð4 ð Ó ðN€4†Ÿ�?�‚Ÿ@ƒ?�„Ÿ@¿�ƒ¿Àÿ ðH ºÈF/ðà Ÿ 𞟨RClick to edit Master text styles Second level Third level Fourth level Fifth level¢!    ª Sðò ð Ó ðN€D“Ÿ�?�‚Ÿ@ƒ?�„Ÿ@¿�ƒ¿Àÿ ðì/º”3ðà Ÿ ð\Ÿ *¡.ø¦ñD¢¢DDå å ‡‡ðô ð Ó ðN€Œ–Ÿ�?�‚Ÿ@ƒ?�„Ÿ@¿�ƒ¿Àÿ ðì/u 1”3ðà  Ÿ ð^Ÿ *¡.ú¦ñD¢¢DDå å ‡‡ðô ð Ó ðN€Ü Ÿ�?�‚Ÿ@ƒ?�„Ÿ@¿�ƒ¿Àÿ ðì/f5ÈF”3ðà Ÿ ð^Ÿ *¡.ئñD¢¢DDå å ‡‡ðH ð ƒ ð0�ƒ“õ!ΔþnF¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ8Š0º___PPT10‹ë.+OÅZ³â ºDefault Designî,0ï € C/ð;/0ð<€ñ ð«.ð( ð ððP ð s 𚀀€ØáŸ�e‚²Žƒe„²Ž…‡¿€‚Í †A‡Áˆ¿ÀË øÿ?¿ÿvesicle“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nð &B 4 ðHŸ¡ ¦ñDææÌ̲²˜˜ð ¢ ð à ðH€@êŸ�e‚²Žƒe„²Ž¿�ÿ‚š¿Àÿ"ñ¿`ðÁ~…þ  ð‡Ÿ¨ÛWhy seek rafts? Animal cell plasma membranes include areas where different species of lipid congregate, these areas are known as rafts. They support specific proteins vital to cell function. Polymersomes are currently used in medicine as microencapsulation to deliver therapeutic substances such as drugs, DNA and proteins. Engineering rafts in artificial membranes would open up new potential applications for polymersomes and make a step towards producing artificial cells.¡,Ì Ìª>¿ #› 5¦ñDææÌ̲²˜˜ðú¢ ð  à ðH€5�e‚²Žƒe„²Ž¿�ƒ¿Àÿðú¤¼ ðPˆHŠ@º___PPT9‹"¬ ð*Ÿ¨>Making polymersomes Dissolve amphiphilic plastic in a good solvent such as tetrahydrofuran (THF) Drip pure water into the solution while stirring: the plastic forms bilayers to keep its hydrophobic chains away from the water. In 80% water, the lowest energy configuration of a bilayer is a closed shell: a polymersome.¡4+! +ªt  #Kh ¦ùDhnÂuÉ|ЃÖð2 ð  S ðŽ€„€`5�0Ç‚0ǃ0Ç„0Ç…‡¿ÍL šY?€†A‡Áˆ¿ÀÿË øÿvesicle"ñ¿``ð -í5N( ðHŸ¡$¦ñDææÌ̲²˜˜ð¦¢ ð  à ðH€à5�e‚²Žƒe„²Ž¿�ƒ¿ÀÿðV >5Hg ð.Ÿ `Copolymers The plastics used in these experiments were PEO45-PMMA180 (polyethylene oxide-polymethyl methacrylate) and Brij-97 (polyoxyethylene-10-oleyl-ether). Both are copolymers - formed by covalently bonding two different polymers  and amphiphilic - one polymer is hydrophilic, the other hydrophobic.¡N1  / çÿ çÿíª>Y  € 6¦ñDææÌ̲²˜˜ð㢠ð 3 ð‚€¸5�e‚²Žƒe„²Ž¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðö03Z% ðóŸ¨The Mini-project Polymersomes (hollow plastic balls) were made from two types of plastic differing in length and composition. The intention was to make rafts in the membrane similar to lipid rafts found in biological cell membranes. The polymersomes produced were hundreds of nanometres in diameter so electron microscopy was used to observe them. Negative staining showed puzzling structures which cryogenic TEM seemed to show were aggregations of smaller vesicles. However, the scanning electron microscope revealed holed membranes. ¡j&N##™þ"# #fþD##ffþ#ª, Ñ !¦ñDææÌ̲²˜˜ðp¢ ð 3 ð‚€DD5�e‚²Žƒe„²Ž¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‰²NHù  ð€Ÿ ÔA polymersome is an artificial vesicle with a bilayer membrane composed of amphiphilic plastic polymers. The term derives from  liposome which denotes a biological vesicle  a hollow ball formed from a phospholipid bilayer membrane.¡,ë Þª> > v ¦ñDææÌ̲²˜˜ððF ð'{®?« ð0 ðð|²©H2ð<¢ ð"  ðÆ€¬5�?�‚Ÿ@ƒ?�„Ÿ@…†‡ˆ‰Š‹¿�‚ƒ„¿ ÀÿÄ˨)ÿ?¿ÿ?„†ˆ3"ñŒ�0e¿`ð'¨o?�ðà  5 ð žð€² ð! c ðB€€A Á¿ÿrichard_wilson"ñ¿`ð.;{®?«ðP¢ ð/ 3 ð‚€œG5�e‚²Žƒe„²Ž¿€†A‡Áˆ¿ ÀË øÿ?¿ÿvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðÂ#ž0¿H½, ð`Ÿ âConclusion Mixing two different polymers has resulted in unusual structures that invite further study and will probably be useful in future plastics technology. However, to produce rafts would seem to require a different combination of polymers. Perhaps this is because Brij-97 by itself doesn t form bilayers: a third species that does could be added or substituted. ¡"r  gª-=¦ñDææÌ̲²˜˜ð ðF ð1Ã9ÿþB ð1 ðð',Yµ/3ðÒ¢ ð ã ðT€ŒV5�e‚²Žƒe„²Ž¿�ƒ¿ÀÿËÎÿˆ"ñ¿`ð1Ã9ËþB ð8Ÿ ^Acknowledgements Supervisor: Dr ir Stefan A F Bon Mentor: Christopher C J Parmenter TEM facilities: Dr Svetla S McPhie Support: MOAC Funding: EPSRC Hitchhiker s Guide pic: BBC¡6ŸŸªb ( 1¦ñDææÌ̲²˜˜ðÚ² ð "  ð’„…�0e‚˜²ƒ0e„˜²AÁ,�ÿÿÿƒÿÿÿ¿Àÿ€€€ˆ_1442161_hitchnew_300ðŸÔ:ÿ ?ðà 5𣢠ð2 ó ðZ€�u5�e‚²Žƒe„²Ž¿¿ ÀÿËÎÿ?¿ÿ“"ñ6@¿ `ÿÀ¿‚‚N¿NÿN?Nð�-g0åHù1 ðÛŸ¨ßReferences Dennis E Discher and Adi Eisenberg, Polymer Vesicles, Science, 2002, 297:967 Wolfgang H Binder, Veronique Barragan and Frederic M Menger, Domains and rafts in Lipid Membranes, Agew. Chem. Int. Ed., 2003, 42, 5802¡X Õ $V$'ªbR(!¦ñDææÌ̲²˜˜ð ðF ðÖõ Åâ5 ð9 ððâY)Ñ)𸲠ð s ðz€€AÁP¿ÿˆpolymerzomes with rafts_75000X_25 small"ñ¿`ðÖ#æN2𺲠ð s ð|€€AÁR¿ÿˆpolymerzomes with rafts_300000X_23 small"ñ¿`ðµB#Å{2ð•¢ ð& C ðˆ€k5�e‚²Žƒe„²Ž¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð‹2 â5 𗟨9scale bar 200 nm scale bar 50 nm Negative stain TEM¡$:'  ™þ¦ñDææÌ̲²˜˜ð”¢ ð3 c 𔀔ƒ5�e‚²Žƒe„²Ž…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nðcõ �Å" ðŠŸ¨Result 1: a weird structure!¡4 ™þ ™þ ¦ñDææÌ̲²˜˜ðJðF ð "077óA ð< ððF%,´-ï2ð¶² ð s ðx€€AÁN¿ÿ ˆ20% Brij polymersomes-32250X-115 small"ñ¿`ðY%Ö1¿3<@𛢠ð* c 𔀘†5�e‚²Žƒe„²Ž…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð¦$$@•4óA 𑟨Cryo-TEM scale bar 100 nm¡$  fþ ª¦ñDææÌ̲²˜˜ð“¢ ð4 c ð”€Ð 5�e‚²Žƒe„²Ž…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð "077Ó1 ð‰Ÿ¨%Result 2: an aggregation of vesicles?¡*& fþ fþ¦ñDææÌ̲²˜˜ð=ðF ðÄCí%VÀ- ð; ððÿ8NGÏ"ð|¢ ð, c ð”€�¬5�e‚²Žƒe„²Ž…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?Nð¾Fð+žSÀ- ðrŸ¨SEM scale bar 100 nm¡$ ffþ ¦ñDææÌ̲²˜˜ðcð@ ðÄCí%V, ð:ðÄCí%V,ð€² ð s ðB€€AÁ¿ÿˆrichard1-81"ñ¿`ð0DÔÒU,ð‹¢ ð5 c ð”€ô¦5�e‚²Žƒe„²Ž…‡¿€†A‡Áˆ¿ ÀË øÿ?¿ÿˆvesicle“"ñ6@¿ ÿÀ¿‚‚N¿NÿN?NðÄCí%V½ ð�Ÿ¨Result 3: perforated membrane¡* ffþ ffþ¦ñDææÌ̲²˜˜ðH ð ƒ ð0�ƒ“õ!ΔþnF¿ÿ ?ð ÿÿÿ€€€»àã33™™™™Ìˆ‘Љº___PPT10‹ië.+OÅ à´â+Dñ='ñ ¹ÿÿÿÿ=ñ @Bñ +r ·,N;êEõm“,v™