Abstract
The influence of high‐pressure freezing (HPF) on the lipid arrangement in phospholipid model membranes has been investigated. Liposomes consisting of pure dipalmitoylphosphatidylcholine (DPPC) and of DPPC mixed with a branched‐chain phosphocholine (1,2‐di(4‐dodecyl‐palmitoyl)‐sn‐glycero‐3‐phosphocholine) have been analysed by freeze‐fracture electron microscopy. The liposomes were frozen either by plunging into liquid propane or by HPF. The characteristic macroripple‐phase of the two‐component liposome system is drastically changed in its morphology when frozen under high‐pressure conditions. The influence of ethanol which acts as pressure transfer medium was ruled out by control experiments. In contrast, no high‐pressure alterations of the pure DPPC bilayer membrane have been observed. We assume that the modification of the binary system is due to a pressure‐induced relaxation of a stressed and unstable lipid molecule packing configuration. HPF was performed with a newly designed sample holder for using sandwiched copper platelets with the high‐pressure freezing machine Balzers HPM010. The sandwich construction turned out to be superior to the original holder system with regard to freeze‐fracturing of fluid samples. By inserting a spacer between the supports samples with a thickness of 20–100 μm can be high‐pressure frozen. The sandwich holder is provided with a thermocouple to monitor cooling rates and allows exact sample temperature control. Despite a two‐fold mass reduction compared to the original holder no HPF cooling rate improvement has been achieved (4000 °C s−1). We conclude that the cooling process in high‐pressure freezing is determined mainly by cryogen velocity.
References
44
Referenced
16
-
Bachmann L.&Mayer E.(1987) Physics of water and ice: implications for cryofixation.Cryotechniques in Biological Electron Microscopy(ed. by R. Steinbrecht and K. Zierold) pp.331. Springer‐Verlag Berlin.
(
10.1007/978-3-642-72815-0_1
) 10.1016/0304-3991(94)90012-4
10.1021/bi00357a047
10.1016/S0006-3495(95)80312-7
10.1016/S0006-3495(91)82072-0
10.1021/bi00271a026
10.1111/j.1365-2818.1987.tb02857.x
10.1021/bi00039a003
10.1111/j.1365-2818.1992.tb01493.x
10.1111/j.1365-2818.1996.tb04793.x
10.1016/0005-2736(80)90331-4
10.1016/0301-4622(95)00085-2
10.1111/j.1365-2818.1994.tb04785.x
10.1046/j.1365-2818.1996.820642.x
10.1016/0009-3084(96)02579-0
10.1016/0005-2736(79)90002-6
10.1046/j.1365-2818.1996.1090666.x
{'key': 'e_1_2_7_19_2', 'first-page': '61', 'article-title': 'Hochdruckgefrieren und Gefrierschneiden von Lecithin‐Liposomen', 'volume': '39', 'author': 'Listemann J.', 'year': '1993', 'journal-title': 'Eur. J. Cell Biol. Suppl'}
/ Eur. J. Cell Biol. Suppl / Hochdruckgefrieren und Gefrierschneiden von Lecithin‐Liposomen by Listemann J. (1993)10.1021/bi00634a030
10.1016/0005-2736(95)00076-F
10.1021/bi00223a023
10.1016/0005-2760(96)00054-9
10.1016/0009-3084(96)02592-3
-
Moor H.(1987) Theory and practice of high‐pressure freezing. Cryotechniques in Biological Electron Microscopy(ed. by R. A. Steinbrecht and K. Zierold) pp.175191. Springer‐Verlag Berlin.
(
10.1007/978-3-642-72815-0_8
) 10.1021/bi00181a001
10.1073/pnas.75.10.4906
{'key': 'e_1_2_7_28_2', 'first-page': '1295', 'article-title': 'Optimization and application of jet‐freezing', 'volume': '7', 'author': 'Müller T.', 'year': '1993', 'journal-title': 'Scanning Microsc'}
/ Scanning Microsc / Optimization and application of jet‐freezing by Müller T. (1993)10.1007/BF01869138
10.1016/0009-3084(86)90012-5
10.1007/BF00576710
10.1111/j.1365-2818.1993.tb03393.x
-
Seddon J.M.&Templer R.H.(1995) Polymorphism of lipid–water systems.Structure and Dynamics of Membranes. Vol. 1A. From Cells to Vesicles(ed. by R. Lipowsky and E. Sackmann) pp.97160. Elsevier Science Amsterdam.
(
10.1016/S1383-8121(06)80020-5
) 10.1016/S0006-3495(94)80790-8
10.1021/bi00714a012
10.1111/j.1365-2818.1995.tb03648.x
10.1016/0301-4622(82)80025-2
10.1073/pnas.93.14.7008
10.1016/0022-2836(73)90303-3
10.1016/0005-2736(74)90023-6
10.1073/pnas.72.1.210
10.1103/PhysRevA.40.2712
{'key': 'e_1_2_7_43_2', 'first-page': '225', 'article-title': 'Effect of compression on rat‐liver mitochondrial membranes. A biochemical and morphological study', 'volume': '23', 'author': 'Wattiaux R.', 'year': '1981', 'journal-title': 'Acta Histochem. Suppl'}
/ Acta Histochem. Suppl / Effect of compression on rat‐liver mitochondrial membranes. A biochemical and morphological study by Wattiaux R. (1981)10.1016/S0006-3495(91)82216-0
10.1126/science.3344420
Dates
Type | When |
---|---|
Created | 22 years, 5 months ago (March 12, 2003, 1:26 a.m.) |
Deposited | 1 year, 11 months ago (Sept. 11, 2023, 1:07 a.m.) |
Indexed | 1 year ago (July 24, 2024, 5:59 a.m.) |
Issued | 27 years, 2 months ago (June 1, 1998) |
Published | 27 years, 2 months ago (June 1, 1998) |
Published Online | 23 years, 7 months ago (Dec. 25, 2001) |
Published Print | 27 years, 2 months ago (June 1, 1998) |
@article{Semmler_1998, title={High‐pressure freezing causes structural alterations in phospholipid model membranes}, volume={190}, ISSN={1365-2818}, url={http://dx.doi.org/10.1046/j.1365-2818.1998.00328.x}, DOI={10.1046/j.1365-2818.1998.00328.x}, number={3}, journal={Journal of Microscopy}, publisher={Wiley}, author={Semmler and Wunderlich and Richter and Meyer}, year={1998}, month=jun, pages={317–327} }