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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1083-1097 |
| Number of pages | 15 |
| Journal | Current Eye Research |
| Volume | 9 |
| Issue number | 11 |
| DOIs | |
| State | Published - 1990 |
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ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
- Cellular and Molecular Neuroscience
Cite this
Methods for the circumvention of problems associated with the study of the ocular Lens plasma membrane-cytoskeleton complex. / FitzGerald, Paul G.
In: Current Eye Research, Vol. 9, No. 11, 1990, p. 1083-1097.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Methods for the circumvention of problems associated with the study of the ocular Lens plasma membrane-cytoskeleton complex
AU - FitzGerald, Paul G
PY - 1990
Y1 - 1990
N2 - 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.
AB - 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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U2 - 10.3109/02713689008997582
DO - 10.3109/02713689008997582
M3 - Article
VL - 9
SP - 1083
EP - 1097
JO - Current Eye Research
JF - Current Eye Research
SN - 0271-3683
IS - 11
ER -