A new method for transfer of polyethylene glycol-embedded tissue sections to silanated slides for immunocytochemistry.
Polyethylene glycol (PEG) is an excellent embedding medium for immunohistochemical studies. It provides structural preservation superior to frozen sections and increased sensitivity of antigen detection compared with paraffin sections. One limitation of PEG embedment is that PEG sections are difficult to handle and adhere poorly to glass slides. Here we present a simple and effective method for embedding tissues in PEG and transferring the resultant sections onto silanated glass slides. In addition, a method for silver enhanced colloidal gold immunostaining was combined with common dye staining to demonstrate the excellent structure preservation and sensitive antigen detection. Bovine chorionic membrane was fixed with Bouin's fixative, embedded in polyethylene glycol (PEG) 1500, cut into 5-microns sections, flattened over agarose blocks (10 x 10 x 2 mm3), and blotted onto Digene silanated slides. Slides were then washed in PBS, which removed the PEG and agarose blocks. Tissue sections were immunocytochemically stained with dilute antiserum raised in a rabbit against purified bovine placental retinol binding protein (bpRBP). Sections were washed and incubated with 1-nm colloidal gold-labeled goat anti-rabbit IgG. The immunogold particles were enhanced by silver staining (IGSS). Specimens were observed and photographed with an Olympus epipolarization microscope. The new method offered excellent morphological preservation of cell structure and the epipolarization microscopy provided high sensitivity for detection of specific immunogold-silver particles.
Gao
K X
KX
Department of Animal Science, University of Tennessee, Knoxville 37901-1071.
Godkin
J D
JD
D000818:Animals; D002417:Cattle; D002823:Chorion / Q000166:cytology / Q000378:metabolism; D005260:Female; D006652:Histological Techniques; D007150:Immunohistochemistry / Q000379:methods*; D011092:Polyethylene Glycols; D012177:Retinol-Binding Proteins / Q000378:metabolism; D012821:Silanes
D012177:Retinol-Binding Proteins; D012821:Silanes; D011092:Polyethylene Glycols