Double-fluorescence image microscopy for quantitation of prostate-specific antigen in histologic sections of the prostate.
BACKGROUND
Although the assessment of serum prostate-specific antigen (PSA) has become a powerful instrument in the diagnosis and for prognosis of prostate carcinoma, there are few quantitative studies of PSA in tissue sections.


METHODS
We developed a technique using double-fluorescence image microscopy for quantifying immunohistochemical reactions in tissue sections. PSA was stained by Texas Red and the cellular DNA was counterstained with 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI). The fluorescence of Texas Red and DAPI was quantified separately after subtraction of background and shading correction. The amount of PSA related to the amount of DNA in identical tissue parts was studied in archival specimens from patients with hyperplasia and prostate carcinoma.


RESULTS
The amount of tissue PSA decreased with the increase in tumor grade, Gleason score, and the change from diploid to aneuploid.


CONCLUSIONS
Double-fluorescence image microscopy is a valuable technique for obtaining quantitative information of cellular constituents. For standardization of immunochemical reactions in tissue sections, cellular DNA seems to be most appropriate.
Department of Medical Radiobiology, Karolinska Institute, 171 76 Stockholm, Sweden.
Naining
N
Wang
Bernhard
B
Tribukait
D006801:Humans; D007211:Indoles / Q000737:chemistry; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods*; D011467:Prostate / Q000737:chemistry* / Q000166:cytology; D017430:Prostate-Specific Antigen / Q000032:analysis* / Q000097:blood / Q000276:immunology; D011471:Prostatic Neoplasms / Q000175:diagnosis*; D013194:Staining and Labeling / Q000379:methods; D014966:Xanthenes / Q000737:chemistry
D007211:Indoles; D014966:Xanthenes; C007293:DAPI; C034657:Texas red; D017430:Prostate-Specific Antigen