Cytometrically coherent transfer of receptor proteins on microporous membranes.
A new method (Freeze-Transfer) is described for performing high-resolution immunocytochemistry for soluble cell proteins on frozen sections of biological tissues that involves thaw-mounting frozen tissue sections directly onto the surface of nitrocellulose thin films instead of directly onto glass slides. This technically straight-forward change in methodology resulted in chromogenic immunocytochemical assays for Her-2 and EGF receptors that were 1-2 orders of magnitude more sensitive while still fully utilizing the diagnostic resolving power of light microscopy. The effects of membrane pore size and surface chemistry on the resolution and intensity of Her-2 signal suggest that the enhanced sensitivity of Freeze-Transfer was caused by the cytologically coherent transfer of target molecules normally lost from cut surfaces of cells mounted on nonporous glass during assay.
McGrath
C M
CM
Grace Bio-Oncology Laboratory, Inc., Pontiac, MI 48342.
Grudzien
J L
JL
Decker
D A
DA
Robbins
T O
TO
D001943:Breast Neoplasms; D003101:Collodion; D066246:ErbB Receptors / Q000032:analysis*; D005615:Freezing; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008567:Membranes, Artificial; D009856:Oncogene Proteins, Viral / Q000032:analysis*; D018719:Erb-b2 Receptor Tyrosine Kinases; D011956:Receptors, Cell Surface / Q000032:analysis*; D012995:Solubility; D013194:Staining and Labeling; D016707:Tissue Fixation
D003101:Collodion; D066246:ErbB Receptors; D008567:Membranes, Artificial; D009856:Oncogene Proteins, Viral; D018719:Erb-b2 Receptor Tyrosine Kinases; D011956:Receptors, Cell Surface