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. 2023 May;40(5):1083-1088.
doi: 10.1007/s10815-023-02764-5. Epub 2023 Mar 17.

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

I-Shen Huang  1   2   3 , Wei-Jen Chen  1   3 , Li-Hua Li  4   5 , Eric Yi-Hsiu Huang  1   3 , Jen-Fan Hang  4 , Chih-Yi Hsu  4 , Chin-Chen Pan  4 , Robert E Brannigan  6 , William J Huang  7   8   9
Affiliations

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

I-Shen Huang et al. J Assist Reprod Genet. 2023 May.

Abstract

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.

Keywords: Azoospermia; Germ cells; Spermatogenesis; Testis; Thionine.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Cytological features of patients diagnosed with A Sertoli cell-only: showing only scattered clusters of Sertoli cell (arrow). B Early maturation arrest: note the absence of spermatid or spermatozoa, while Sertoli cell and primary spermatocyte (arrowhead) continue to be prominent. C Late maturation arrest: spermatids (hashtag) are detected but without spermatozoa. D Complete spermatogenesis: the asterisk indicates mature spermatozoa. (Thionine stain, reduced from × 400)
Fig. 2
Fig. 2
Concordance of TPS and histopathology in relation to individual germ cell types recognized by histopathology in relation to TPS findings
See this image and copyright information in PMC

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