Raman spectroscopy as an ex vivo noninvasive approach to distinguish complete and incomplete spermatogenesis within human seminiferous tubules

Raman spectroscopy may noninvasively identify different maturational stages of spermatogenesis in ex vivo human seminiferous tubules.

Yufei Liu, M.D., Zheng Li, M.D., Yong Zhu, Ph.D., Ling Di, Ph.D., Feng Liu, B.S., Lin He, Ph.D., Hongliang Hu, Ph.D., E. Charles Osterberg, M.D., Yiran Huang, M.D., Philip S. Li, M.D.

Volume 102, Issue 1, Pages 54–60.e2


To evaluate the potential clinical application of Raman spectroscopy (RS) as a tool that may identify spermatogenesis within human seminiferous tubules.

RS scanning of human testicular tissue at different maturational stages; immunohistochemistry study and metabolomic analysis of nonobstructive azoospermic/obstructive azoospermic testes.

State-owned hospital.

Fifty-two patients with clinical indications of nonobstructive azoospermia (NOA) and obstructive azoospermia (OA) who underwent infertility evaluation and treatment.


Main Outcome Measurement(s):
Raman spectra of seminiferous tubules, thickness of lamina propria (LP), immunohistochemistry of type I, III, and IV collagens and laminin, metabolites of human testes.

Tubules of OA patients had spectral intensities below 2,000 (au), while tubules of NOA patients had higher intensities, depending on the degree of spermatogenesis. RS was able to separate samples of NOA and OA testicular tissue with a sensitivity of 90% and specificity of 85.71%. The LP of NOA tubules were thickened and had increased deposition of type I and type III collagens. Gas chromatography-mass spectrometer (GC-MS) detected 12 metabolites that showed significant differences between NOA and OA testes.

RS can noninvasively distinguish seminiferous tubules with complete and incomplete spermatogenesis and may serve as a novel and potentially useful tool to guide surgeons performing micro-testicular sperm extraction to improve sperm retrieval.

  • Jason Kovac

    I agree with both comments below. I would think that the cost is obviously prohibitive currently but does anyone have any clue as to whether this will ever be something commonly available? I would think that the trouble in our field is that relatively speaking the patient volume is so little, making it difficult to market technologies specifically for the treatment of infertility.

  • Interesting technology. While it’s impressive to be able to differentiate seminiferous tubules from NOA and OA patients, it is more clinically relevant to be able to identify tubules with spermatogenesis in NOA patients. Any advancement to improve microTESE outcome is much needed.

  • This study builds upon the data initially presented by Dr. Schegel several years ago initially at the ASRM meeting. Any technology that can improve our retrieval rates for MicroTESE procedures will help benefit the couple. As this technology progresses, my hope is that more objective data can be used to guide male infertility specialists in this highly technical procedure. Well done.

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