Fibrin clots keep non-adhering living cells in place on glass for perfusion or fixation.
We describe a method to hold living cells in place that ordinarily do not adhere to glass coverslips. The method, developed for insect spermatocytes but with application to other cell types, consists of embedding cells in a fibrin clot that forms after the enzyme thrombin cleaves the blood protein fibrinogen. The method permits continuous observation of living cells as they are treated with and recover from drug or other treatments: when held in the clot the living cells remain in place and keep their shapes when perfused with drugs that ordinarily cause drastic shape changes, and they remain in place and keep their shapes through lysis/fixation procedures. We describe how to place live cells in a fibrin clot and how subsequently to perfuse them.
Forer
Arthur
A
Biology Department, York University, Toronto, Ontario, Canada M3J 1P3. aforer@yorku.ca
Pickett-Heaps
Jeremy
J
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D004175:Diptera / Q000166:cytology; D005337:Fibrin; D005898:Glass; D007552:Isotonic Solutions / Q000737:chemistry; D008297:Male; D008853:Microscopy; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D013090:Spermatocytes / Q000166:cytology; D016610:Tissue Embedding / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods*
D007552:Isotonic Solutions; D005337:Fibrin