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Improved cryoprotection and freeze-substitution of embryonic quail retina: a TEM study on ultrastructural preservation

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Meissner,  DH
Department Cell Biology, Max Planck Institute for Developmental Biology, Max Planck Society;

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Schwarz,  H
Electron Microscopy, Max Planck Institute for Developmental Biology, Max Planck Society;

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Citation

Meissner, D., & Schwarz, H. (1990). Improved cryoprotection and freeze-substitution of embryonic quail retina: a TEM study on ultrastructural preservation. Journal of Electron Microscopy Technique, 14(4), 348-356. doi:10.1002/jemt.1060140410.


Cite as: https://hdl.handle.net/21.11116/0000-000F-234A-7
Abstract
Conditions for cryofixation and freeze-substitution crucial to the ultrastructural preservation of embryonic quail retina were improved. As freeze-substitution makes gentle dehydration and chemical fixation of tissue possible, the suitability of different cryoprotectants were tested in the preceding cryofixation. Additionally, different conditions for chemical prefixation were studied. In cryofixation, all of the "classic" cryoprotectants caused more or less severe tissue destruction. Only dimethylformamide (DMF) and--with certain reservations--dimethylsulfoxide (DMSO) yielded improved structure preservation. Perfusion fixation with a mixture of formaldehyde/glutaraldehyde (FA/GA) was superior to GA alone. In comparison to conventional fixation and dehydration methods, freeze-substitution yielded better ultrastructural preservation of the embryos with fewer artifacts.