Raman spectroscopy as an ex vivo noninvasive approach to distinguish complete and incomplete spermatogenesis within human seminiferous tubules.
OBJECTIVE
To evaluate the potential clinical application of Raman spectroscopy (RS) as a tool that may identify spermatogenesis within human seminiferous tubules.


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


METHODS
State-owned hospital.


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


METHODS
None.


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


RESULTS
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.


CONCLUSIONS
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.
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China; Shanghai Human Sperm Bank, Shanghai, People's Republic of China.
Yufei
Y
Liu
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China; Shanghai Human Sperm Bank, Shanghai, People's Republic of China.
Yong
Y
Zhu
Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Ling
L
Di
Center for Male Reproductive Medicine and Microsurgery, Department of Urology, Weill Cornell Medical College, New York, New York.
E Charles
EC
Osterberg
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China; Shanghai Human Sperm Bank, Shanghai, People's Republic of China.
Feng
F
Liu
Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
Lin
L
He
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China; Shanghai Human Sperm Bank, Shanghai, People's Republic of China.
Hongliang
H
Hu
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China; Shanghai Human Sperm Bank, Shanghai, People's Republic of China.
Yiran
Y
Huang
Center for Male Reproductive Medicine and Microsurgery, Department of Urology, Weill Cornell Medical College, New York, New York.
Philip S
PS
Li
Department of Urology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, People's Republic of China; Shanghai Human Sperm Bank, Shanghai, People's Republic of China. Electronic address: Lizhengboshi@163.com.
Zheng
Z
Li
D053713:Azoospermia / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D015415:Biomarkers / Q000032:analysis; D003094:Collagen / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin / Q000032:analysis; D008297:Male; D055432:Metabolomics / Q000379:methods; D011237:Predictive Value of Tests; D012671:Seminiferous Tubules / Q000737:chemistry / Q000473:pathology* / Q000503:physiopathology; D013059:Spectrum Analysis, Raman; D013091:Spermatogenesis; D013094:Spermatozoa / Q000737:chemistry / Q000473:pathology*
D015415:Biomarkers; D007797:Laminin; D003094:Collagen
GC-MS; Raman spectroscopy; human seminiferous tubules; immunohistochemistry; sperm retrieval