An innovative fixative for cytoskeletal components allows high resolution in colocalization studies using immunofluorescence techniques.
Using a new fixation solution, CytoSkelFix, it is now possible to obtain superior fixation and thus resolution of cytoskeletal components using immunofluorescence and fluorescence microscopy. This fixative combines rapid cell penetration and cellular crosslinking of proteins such that both preservation and resolution of cellular proteins can be detected. The cytoskeleton has proven very difficult to preserve, partly because of the lability of one of the filament systems (microtubules), and one single fixative is incapable of properly preserving microtubules, microfilaments, and intermediate filaments for localization in the same cell. Further, the motor proteins associated with the cytoskeletal elements are even more difficult to preserve, particularly simultaneously with the fiber system with which they associate. We present evidence that CytoSkelFix is a superior preservative and would be useful in fixation for all types of immunofluorescence colocalization studies where superior preservation is required.
Robinson
Rollin W
RW
University of Denver, 2190 E. Iliff Avenue, CO 80208, Denver, USA.
Snyder
Judith A
JA
D000199:Actins / Q000032:analysis; D000818:Animals; D015415:Biomarkers; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000737:chemistry; D004398:Dyneins / Q000032:analysis; D005404:Fixatives; D019084:Fluorescent Antibody Technique, Indirect; D016547:Kinesins / Q000032:analysis; D008870:Microtubules / Q000737:chemistry / Q000648:ultrastructure*; D009218:Myosins / Q000032:analysis; D048328:Potoroidae; D016707:Tissue Fixation / Q000379:methods*; D014404:Tubulin / Q000032:analysis
D000199:Actins; D015415:Biomarkers; D003432:Cross-Linking Reagents; D005404:Fixatives; D014404:Tubulin; D009218:Myosins; D004398:Dyneins; D016547:Kinesins