Background: Azoospermia is the most severe form of male infertility, affecting approximately 1% of male population and 10–15% of infertile men. In azoospermia cases, sperm retrieval from the testis or epididymis through surgical procedures is used for assisted reproductive treatments. When no sperm is retrieved, recent approaches in medicine suggest using immunohistochemical methods to evaluate spermatogenesis in testicular tissue and plan further treatments. Aim: To evaluate spermatogenesis in testicular tissue of azoospermic patients using immunohistochemistry and compare the findings with the clinical outcomes of sperm retrieval procedures. Materials and Methods: This study included 71 azoospermic men who underwent micro-TESE procedures at the IVF center (RMC) between 2019 and 2023. The excised testicular tissues were fixed, processed histologically, and evaluated using Johnson’s score. The presence of spermatozoa in seminiferous tubules was detected by immunohistochemical and immunofluorescence staining, using markers such as TEX101 and LDHC. Results: Johnson’s score was categorized into three groups: poor, moderate, and good spermatogenesis. These were statistically compared with hormonal levels and surgical sperm retrieval outcomes. There were significant differences in Johnson’s scores and serum FSH and LH levels among the three groups (p < 0.005). TEX101 and LDHC proteins were strongly expressed in the good group, weakly in the moderate group, and absent in the poor group. The success rate of sperm retrieval was 100% (17/17) in the good group, 96.29% (26/27) in the moderate group, and only 29.62% (8/27) in the poor group. Conclusion Histological evaluation of spermatogenesis in azoospermic patients can help predict the outcome of surgical sperm retrieval, indicating its clinical value in treatment planning.