IMPROVED CRYOPROTECTION AND FREEZE-SUBSTITUTION OF EMBRYONIC QUAIL RETINA - A TEM STUDY ON ULTRASTRUCTURAL PRESERVATION
IMPROVED CRYOPROTECTION AND FREEZE-SUBSTITUTION OF EMBRYONIC QUAIL RETINA - A TEM STUDY ON ULTRASTRUCTURAL PRESERVATION
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DOI：
10.1002/jemt.1060140410
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发表时间：
1990-04-01
期刊：
影响因子：
--
通讯作者：
SCHWARZ, H
中科院分区：
文献类型：
--
作者：
MEISSNER, DH;SCHWARZ, H
改进了对胚胎视网膜超微结构保存至关重要的冷冻固定和冷冻替代条件。由于冷冻替代使组织的温和脱水和化学固定成为可能，因此在之前的冷冻固定中测试了不同冷冻保护剂的适用性。此外，还对不同的化学前固定条件进行了研究。在冷冻固定中，所有经典的冷冻保护剂都或多或少地造成了严重的组织破坏。只有二甲基甲酰胺(DMF)和二甲基亚砜(DMSO)--尽管有一定的保留--能改善结构保持性。甲醛/戊二醛混合液(FA/GA)灌流固定优于单独使用GA。与传统的固定和脱水方法相比，冷冻替代法能更好地保存胚胎的超微结构，伪影较少。
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.