TESTIS BIOPSY IMAGE-ANALYSIS EFFECTIVELY QUANTIFIES SPERMATOGENIC CELL-TYPES

Purpose: Evaluation of the testis biopsy specimen in male infertility has been hampered by the qualitative rather than quantitative nature o f routine histological techniques and interpretation. Although useful, now cytometry has demonstrated significant limitations due to its ina bility to differentiate among haploid cell types, spermatozoa and sper matids. Thus, deoxyribonucleic acid image analysis was performed on te stis biopsy specimens to combine the advantages of quantitative ploidy and morphological characteristics to develop a more effective quantit ative system. Materials and Methods: Image analysis using a specifical ly designed filter was performed on Feulgen stained 5 mu m. sections o f paraffin embedded testicular tissue. The archival testicular tissue had been obtained using standard biopsy techniques from patients with azoospermia and normal spermatogenesis on routine hematoxylin and eosi n processed tissue. Results: The overall findings (mean plus or minus standard deviation) in 20 biopsies from 18 men are haploid 54.2 +/- 5. 6, diploid 31.2 +/- 5.2 and tetraploid 15.0 +/- 4.6%. Spermatozoa and spermatids contributed 23.2 +/- 5.7 and 30.9 +/- 7.6% to the total cel l content, respectively. A mean of 2,462 +/- 254 cells were counted pe r case. Conclusions: We used image analysis to evaluate testicular bio psies. This technique can quantitatively determine the percentages of various cell types within the seminiferous tubules. Furthermore, it ca n differentiate between spermatozoa and spermatids based on morphologi cal characteristics. This technique, which combines ploidy and cell mo rphology characteristics, represents a significant advancement in the interpretation of the testicular biopsy, and has implications for rede fining the presently used system of qualitative interpretative assessm ent.