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This trial was prepared on ITO plates instead of glass slides
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This trial was prepared on ITO plates instead of glass slides.
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As new liposomes developed and swelled, previously formed liposomes were often pushed away from the surface. The liposomes in the resulting layers may be tethered to the surface or to other liposomes some tethers were visible when viewed by phase contrast microscopy
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As new liposomes developed and swelled, previously formed liposomes were often pushed away from the surface. The liposomes in the resulting layers may be tethered to the surface or to other liposomes (some tethers were visible when viewed by phase contrast microscopy).
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A few liposomes appeared to have detached completely but this occurrence was not common within any given population of liposomes. We procured free-floating liposomes by prying apart the chamber and allowing the contents to drip into a collection vessel or by applying gentle suction using a pipette or needle and syringe to remove the solution from the chamber see Supporting Information, Figure S2
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A few liposomes appeared to have detached completely but this occurrence was not common within any given population of liposomes. We procured free-floating liposomes by prying apart the chamber and allowing the contents to drip into a collection vessel or by applying gentle suction using a pipette or needle and syringe to remove the solution from the chamber (see Supporting Information, Figure S2).
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This low yield may be due, in part, because we formed these liposomes without a prehydration step
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This low yield may be due, in part, because we formed these liposomes without a prehydration step.
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To eliminate signal from the autofluorescence of agarose, we first viewed a film of agarose without lipids and adjusted the camera settings until we could no longer observe fluorescence from the agarose film. These settings were then used to view the agarose film with fluorescently labeled lipids; its fluorescence was significantly stronger and easily detectable
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To eliminate signal from the autofluorescence of agarose, we first viewed a film of agarose without lipids and adjusted the camera settings until we could no longer observe fluorescence from the agarose film. These settings were then used to view the agarose film with fluorescently labeled lipids; its fluorescence was significantly stronger and easily detectable.
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We characterized the surface of films prepared from four types of agarose by AFM and SEM (see Supporting Information. Figure S4) and found that films from ultralow melting agarose appeared to display the smoothest (i.e, the least fibrous looking) surface. We attribute this observation to two reasons:(1, the average molecular weight of ultralow melting agarose was the lowest of the four types of agarose (Normand, V, Lootens, D. L, Amici, E, Plucknett, K. P, Aymard, P. Biomacromolecules 2000, 1, 730-738, and (2) ultralow melting temperature agarose did not gel to a noticeable extent during the formation of the agarose film at 40 °C Figure 2, instead, it remained dissolved in solution
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We characterized the surface of films prepared from four types of agarose by AFM and SEM (see Supporting Information. Figure S4) and found that films from ultralow melting agarose appeared to display the smoothest (i.e., the least fibrous looking) surface. We attribute this observation to two reasons:(1). the average molecular weight of ultralow melting agarose was the lowest of the four types of agarose (Normand, V.; Lootens, D. L.; Amici, E.; Plucknett, K. P.; Aymard, P. Biomacromolecules 2000, 1, 730-738.) and (2) ultralow melting temperature agarose did not gel to a noticeable extent during the formation of the agarose film at 40 °C (Figure 2); instead, it remained dissolved in solution.
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Itagaki et al. provide additional evidence of water molecules co- crystallizing with agarose by use of a fluorescent probe (Itagaki, H.; Fukiishi. Ft.; Imai. T.; Watase. M. J. Pol mi. Set, Part B: Polym. Phys, 2005, 43, 680-688).
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Itagaki et al. provide additional evidence of water molecules co- crystallizing with agarose by use of a fluorescent probe (Itagaki, H.; Fukiishi. Ft.; Imai. T.; Watase. M. J. Pol mi. Set, Part B: Polym. Phys, 2005, 43, 680-688).
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Arndt and Stevens discuss the presence of tightly bound water molecules within dried agarose gels as determined by vacuum UV CD Arndt, E. R, Stevens, E. S. Biopolymers 1994, 341527-1534
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Arndt and Stevens discuss the presence of tightly bound water molecules within dried agarose gels as determined by vacuum UV CD (Arndt, E. R.; Stevens, E. S. Biopolymers 1994, 341527-1534).
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In fact, even after 2 min of hydration, high melting agarose appeared to remain as a dry film
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In fact, even after 2 min of hydration, high melting agarose appeared to remain as a dry film.
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We followed the swelling by adjusting the focal plane of the microscope as the hybrid film of agarose and lipids moved away from the surface of die glass slide
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We followed the swelling by adjusting the focal plane of the microscope as the hybrid film of agarose and lipids moved away from the surface of die glass slide.
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0001378233
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One may ask if liposomes grown from hybrid films of agarose and lipid formulations that contained negatively charged lipids or PEGylated lipids (Figure 3C, E, G, M, O, Q, S) also fused during the formation process. In this context, it is instructive that in other methods of formation, such as electroformation or freeze-and-thaw, fusion of adjacent liposome membranes is also a key characteristic in formation of giant liposomes Menger, F. M, Angelova. M. I. Acc. Chem. Res, 1998, 31, 789-797
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One may ask if liposomes grown from hybrid films of agarose and lipid formulations that contained negatively charged lipids or PEGylated lipids (Figure 3C, E, G, M, O, Q, S) also fused during the formation process. In this context, it is instructive that in other methods of formation, such as electroformation or freeze-and-thaw, fusion of adjacent liposome membranes is also a key characteristic in formation of giant liposomes (Menger, F. M.; Angelova. M. I. Acc. Chem. Res, 1998, 31, 789-797.
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Oku, N, Macdonald, R. C. Biochemistry 1983, 22 855-863 and these methods also yield giant liposomes from lipid compositions that include PEGylated lipids, Constantinescu, I, Levin, E, Gyongyossy-lssa, M. Artif. Cells Blood Substit. Immobil. Biotech- nol, 2003, 31, 395-424) or anionic lipids, Claessens, M, Leermakers, F. A. M, Hoeksfra, F. A, Stuart, M. A. C. J. Phys. Chem, B2007, 111 7127-7132, Therefore, the repulsive forces generated by these specialized lipid formulations, which are often used to promote initial bilayer orientation and separation, can be overcome by appropriate mechanical stresses, Angelova, M. 1, Dimitrov, D. S. Prog. Colloid Polym. Set 1988, 7659-67. Cevc, G, Richardsen, H. Adv. DnigDeliv. Rev, 1999, 38 207-232 such as those generated by an electric field, osmotic pressure during freeze-and-thaw cycles, or as proposed here, by a swelling hydrogel film
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Oku, N.; Macdonald, R. C. Biochemistry 1983, 22 855-863 and these methods also yield giant liposomes from lipid compositions that include PEGylated lipids. (Constantinescu, I.; Levin, E.; Gyongyossy-lssa, M. Artif. Cells Blood Substit. Immobil. Biotech- nol, 2003, 31, 395-424) or anionic lipids. (Claessens, M.; Leermakers, F. A. M.; Hoeksfra, F. A.; Stuart, M. A. C. J. Phys. Chem, B2007, 111 7127-7132). Therefore, the repulsive forces generated by these specialized lipid formulations, which are often used to promote initial bilayer orientation and separation, can be overcome by appropriate mechanical stresses. (Angelova, M. 1.; Dimitrov, D. S. Prog. Colloid Polym. Set 1988, 7659-67. Cevc, G.; Richardsen, H. Adv. DnigDeliv. Rev, 1999, 38 207-232) such as those generated by an electric field, osmotic pressure during freeze-and-thaw cycles, or as proposed here, by a swelling hydrogel film.
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A definite analysis of lamellarity is typically obtained by micropipette aspiration to measure the bending modulus Needham. D, Mcintosh. T. J, Lasic, D. D. Bjochim. Biophys. Actal992, 1108, 40-48. Akashi, K, Miyata, H, ltoh, H, Kinosita, K. Biophys. J, 1996, 71, 3242- 3250
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A definite analysis of lamellarity is typically obtained by micropipette aspiration to measure the bending modulus (Needham. D.; Mcintosh. T. J.; Lasic, D. D. Bjochim. Biophys. Actal992, 1108, 40-48. Akashi, K.; Miyata, H.; ltoh, H.; Kinosita, K. Biophys. J, 1996, 71, 3242- 3250.
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Angelova, M.; Soleau, S.; Meleard, P.; Faucon, J. F.; Bothorel, P. Prog. Colloid Polym. Sci, 1992, 89, 127-131. Bermudez, H.;Hammer, D. A.;Discher, D. E. Lang- muir2004, 20, 540-543.
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Angelova, M.; Soleau, S.; Meleard, P.; Faucon, J. F.; Bothorel, P. Prog. Colloid Polym. Sci, 1992, 89, 127-131. Bermudez, H.;Hammer, D. A.;Discher, D. E. Lang- muir2004, 20, 540-543).
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76
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Measuring the bending modulus by micropipet aspiration is difficult to perform and requires specialized equipment and expertise Hotani, H, Nomura, R; Suzuki, Y. Can. Opin. Colloid Interface Sci, 1999, 4, 358-368
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Measuring the bending modulus by micropipet aspiration is difficult to perform and requires specialized equipment and expertise (Hotani, H.; Nomura, R; Suzuki, Y. Can. Opin. Colloid Interface Sci, 1999, 4, 358-368.
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77
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0029752766
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Akashi, K.;Miyata. H.:Itoh. H.;Kinosita, K. Biophys. J, 1996, 71, 3242-3250.
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(1996)
Biophys. J
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, pp. 3242-3250
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Akashi, K.1
Miyata, H.2
Itoh, H.3
Kinosita, K.4
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78
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0037066601
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Yamashita, Y.;Oka, M.;Tanaka, T.;Yamazaki, M. Biochim. Biophys. Acta2002, 1561. 129-134.
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(2002)
Biochim. Biophys. Acta
, vol.1561
, pp. 129-134
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Yamashita, Y.1
Oka, M.2
Tanaka, T.3
Yamazaki, M.4
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79
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3042593664
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Bermudez, H.;Ham- mer, D. A.;Discher, D. E. Langmuir2004, 20, 540-543.
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Bermudez, H.;Ham- mer, D. A.;Discher, D. E. Langmuir2004, 20, 540-543.
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81
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84924210747
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The bending modulus can be affected by ions in the solution (Dimova, R.; Aranda, S.; Bezlyepkina, N.; Nikolov, V.; Riske. K. A.; Lipowskv, R. J. phys.: Condens. Matter2006. 18, S1151-S1176) by macromol- ecules associated with the membrane, such as PEG. (Yamashita, Y.; Oka, M.; Tanaka, T.; Yamazaki, M. Biochim. Biophys. Acta2002,1561, 129-134. Bermudez, Ft.; Hammer, D. A.; Discher, D. E. Langmuir200420, 540-543), or possibly by agarose.
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The bending modulus can be affected by ions in the solution (Dimova, R.; Aranda, S.; Bezlyepkina, N.; Nikolov, V.; Riske. K. A.; Lipowskv, R. J. phys.: Condens. Matter2006. 18, S1151-S1176) by macromol- ecules associated with the membrane, such as PEG. (Yamashita, Y.; Oka, M.; Tanaka, T.; Yamazaki, M. Biochim. Biophys. Acta2002,1561, 129-134. Bermudez, Ft.; Hammer, D. A.; Discher, D. E. Langmuir200420, 540-543), or possibly by agarose.
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