A non-enzymatic method for lens decapsulation which leaves the epithelial cells attached to the fibers.
We have developed a method to mechanically decapsulate the lens yet leave the epithelial cells attached to the fibers. This method uses divalent ion chelaters to loosen the capsule-epithelial interactions and bumetanide to control cell swelling. Light microscopy, scanning electron microscopy, and in-vitro fluorescence microscopy demonstrate that the capsule is removed and the epithelial cells remain adherent to the fibers when these procedures are used. Ion channel activity and epithelial gap junction communication remain following decapsulation. This decapsulated lens preparation should prove useful for many kinds of lens studies.
Dewey
J
J
Department of Physiology and Biophysics, Mayo Foundation, Rochester, MN 55905, USA.
Bartling
C
C
Rae
J L
JL
D000818:Animals; D002417:Cattle; D002469:Cell Separation / Q000379:methods*; D002642:Chick Embryo; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry / Q000648:ultrastructure; D005452:Fluoresceins / Q000032:analysis; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents; D007903:Lens Capsule, Crystalline / Q000166:cytology*; D007908:Lens, Crystalline / Q000166:cytology*; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D012996:Solutions; D013552:Swine
D005452:Fluoresceins; D007202:Indicators and Reagents; D012996:Solutions; C007740:fluorexon