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Ultrastructure of the frog retina after high-pressure freezing and freeze substitution


, : Ultrastructure of the frog retina after high-pressure freezing and freeze substitution. Journal of Microscopy (Oxford) 189(3): 219-235, March

In many types of tissue, high-pressure freezing (HPF), followed by freeze substitution, can produce excellent ultrastructural preservation at depths over 10 times that obtained by other cryofixation techniques. However, in the case of neural tissue, the benefits of HPF have not been realized. In the present study, isolated frog (Rana pipiens) retina was sliced at a thickness of 150 or 350 mum, rapidly frozen in a Balzers HIM 010 high-pressure freezer, and 15% low-MW dextran for 350-mum slices) were required for freezing media (35% high-MW dextran for 150-mum slices or adequate freezing. The quality of preservation after HPF was excellent throughout the retina in both the 150- and 350-mum slices, compared with chemically fixed slices. Specifically, HPF resulted in better preserved cellular, mitochondrial and nuclear membranes in all retinal layers. This is the first study to successfully cryofix all of the layers of the retina. The increased depths of adequate freezing achieved by HPF should facilitate various ultrastructural studies of retina, as well as of other CNS tissues, where preservation approaching that of the 'native' state is required.

US$19.90

PMID: 9588021

DOI: 10.1046/j.1365-2818.1998.00315.x


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