A quantitative assessment by flow cytometry of the preservation of the Thy-1 antigen by various fixation procedures.
A quantitative assessment of the effects of various fixatives upon the preservation of the Thy-1 surface antigen has been undertaken. A panel of 5 monoclonal antibodies (MAbs) have been reacted with mouse lymphocytes treated with various fixatives, and their binding analysed by flow cytometry using a fluorescence activated cell sorter. It was found that 2% paraformaldehyde (PFA) provided the best antigenic preservation, as detected by both the anti Thy-1.1 and anti Thy-1.2 MAbs. The effectiveness of 2% PFA fixation is further demonstrated by the immunohistochemical identification of the Thy-1 antigen in sections of mouse cerebellar cortex.
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D005404:Fixatives; D005434:Flow Cytometry; D005557:Formaldehyde; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D011108:Polymers; D018800:Thy-1 Antigens; D013950:Thymus Gland / Q000032:analysis* / Q000166:cytology; D014021:Tissue Preservation / Q000379:methods*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005404:Fixatives; D011108:Polymers; D018800:Thy-1 Antigens; D005557:Formaldehyde; C003043:paraform