Methods for the circumvention of problems associated with the study of the ocular lens plasma membrane-cytoskeleton complex.

Abstract:

:Two alternative methods for the study of the lens cytoskeleton are described which serve to overcome some of the difficulties imparted by the unique biology of the lens. The first technique involves rapid freezing, thick sectioning, and selective extraction and/or fixation of the lens section. This approach offers several advantages: 1) enhanced visualization of the cytoskeleton, 2) avoidance of fixation gradients, 3) free access for immunocytochemical probes, 4) retention of tissue-wide spatial relationships, with a sharp increase in the resolution of regional analysis, and 5) the capacity for correlative morphological and biochemical comparisons. The second method involves the covalent immobilization of the plasma membrane-cytoskeleton complex (PMCC) to acrylamide beads. This approach permits: 1) avoidance of fixation in the immunocytochemical analysis of lens cytoskeleton and plasma membranes 2) rapid processing of multiple, small-quantity samples for immunocytochemistry/biochemical analysis 3) cleaner and more rapid analysis of cytoskeletal extraction conditions. Both approaches, while particularly suited to the study of the lens PMCC, may also be of value to the study of the PMCC of other tissues, particularly where preservation/analysis of regional relationships is essential.

journal_name

Curr Eye Res

journal_title

Current eye research

authors

FitzGerald PG

doi

10.3109/02713689008997582

subject

Has Abstract

pub_date

1990-11-01 00:00:00

pages

1083-97

issue

11

eissn

0271-3683

issn

1460-2202

journal_volume

9

pub_type

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