An immunohistochemical method for the detection of proteins in the vertebrate lens

Fluorescence immunohistochemistry has traditionally been difficult or impos sible to perform on the vertebrate lens because of its extremely high prote in content. Described here is a robust and rapid method for preparing and l abeling vertebrate eyes for confocal microscopy. This technique has success fully been applied to localize proteins in the lens epithelium and capsule. as well as the primary and secondary fibers. This technique preserves tiss ue morphology and coupled with double and triple labeling. has allowed loca lization of proteins bound to plasma membrane, basement membrane, nucleus, endoplasmic reticulum as well as sub-nuclear compartments. In addition, the present technique has proven useful for fluorescent immunohistochemical an alysis of diverse tissues including whole embryos, adult muscle, pancreas, and liver. This procedure allowed us to successfully localize a wide variet y of antigens on diverse vertebrate tissues including the more challenging vertebrate lens. (C) 2001 Elsevier Science B.V. All rights reserved.