Touch print smear of testicular tissue with thionine stain for intraoperative diagnosis in azoospermic men

I. Shen Huang, Wei Jen Chen, Li Hua Li, Eric Yi Hsiu Huang, Jen Fan Hang, Chih Yi Hsu, Chin Chen Pan, Robert E. Brannigan, William J. Huang*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Purpose: To compare the diagnostic value of testicular tissue touch print smear (TPS) conducted on azoospermic patients with results obtained from histopathology and in vitro fertility (IVF) lab findings. Methods: Microdissection testicular sperm extraction was performed on a group of 148 azoospermic patients and testicular samples obtained intraoperatively. Using TPS, the samples were smeared onto a sterile slide, followed with staining using thionine. The testis tissue bulk samples were also transferred to the IVF lab, and determinations of sperm presence or absence obtained from IVF lab tests were compared with the TPS sample results. Needle testis biopsy was separately performed on a group of 360 azoospermic patients, and results of pathohistology review on the biopsies were further compared with determinations of spermatogenesis stage obtained from TPS for those patients. Results: When compared with IVF lab results, TPS was found to have 100% (126/126) positive predictive value and 95.5% (25/26) negative predictive value for predicting sperm presence or absence, respectively. Furthermore, TPS was further found to have a 93.6% correlation (337 of 360 biopsies) with results of histological diagnoses performed by needle biopsy. Results from histology and TPS for the detection of sperm presence were concordant in 96.1% (346/360) of biopsies. Diagnosis of SCO by TPS shows the highest correlation with histopathology (98.6%), followed by complete spermatogenesis (97.5%), early maturation arrest (78.9%), and late maturation arrest (27.3%). Conclusions: The results support the continued use of TPS in testicular tissue analysis for more rapid assessment of spermatogenesis and for detection of spermatozoa in azoospermic subjects.

Original languageEnglish (US)
Pages (from-to)1083-1088
Number of pages6
JournalJournal of Assisted Reproduction and Genetics
Issue number5
StatePublished - May 2023


  • Azoospermia
  • Germ cells
  • Spermatogenesis
  • Testis
  • Thionine

ASJC Scopus subject areas

  • Reproductive Medicine
  • Genetics
  • Obstetrics and Gynecology
  • Developmental Biology
  • Genetics(clinical)


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