Freeze-fracture enzyme cytochemistry: application of enzyme cytochemistry to freeze-fracture cytochemistry.
We present a novel freeze-fracture cytochemistry method based upon enzyme cytochemistry. By this method, freeze-fractured membranes are labeled with cerium as an enzyme cytochemical marker on replicas. The cerium capture method was suitable for freeze-fracture enzyme cytochemistry because the cerium phosphate reaction product is stable after replica-cleaning. As a model system, acid phosphatase, which is a well-known lysosomal marker, was detected on freeze-fractured membranes of lysosomes in the proximal tubular epithelium of the rat kidney. This technique should be a useful addition for analyzing the ultrastructure of biological membranes.
Takizawa
T
T
Department of Anatomy, Jichi Medical School, Tochigi, Japan.
Saito
T
T
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002563:Cerium; D004798:Enzymes / Q000378:metabolism*; D005614:Freeze Fracturing / Q000379:methods*; D006651:Histocytochemistry / Q000379:methods*; D007668:Kidney / Q000201:enzymology / Q000648:ultrastructure; D008247:Lysosomes / Q000201:enzymology / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley
D004798:Enzymes; D002563:Cerium; D000135:Acid Phosphatase