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Methods for the Circumvention of Problems Associated with the Study of the Ocular Lens Plasma Membrane-Cytoskeleton Complex

 

作者: FitzgeraldPaul G.,  

 

期刊: Current Eye Research  (Taylor Available online 1990)
卷期: Volume 9, issue 11  

页码: 1083-1097

 

ISSN:0271-3683

 

年代: 1990

 

DOI:10.3109/02713689008997582

 

出版商: Taylor&Francis

 

数据来源: Taylor

 

摘要:

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.

 

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