Identification of spermatogenesis in individual seminiferous tubules and testicular tissue of adult normal and busulfan-treated mice employing Raman spectroscopy and principal component analysis

Anand M Shrivastav; Nagham Ali; Neetika Singh; Eitan Lunenfeld; Ibrahim Abdulhalim; Mahmoud Huleihel

doi:10.1016/j.saa.2024.124232

Identification of spermatogenesis in individual seminiferous tubules and testicular tissue of adult normal and busulfan-treated mice employing Raman spectroscopy and principal component analysis

Spectrochim Acta A Mol Biomol Spectrosc. 2024 Jul 5:315:124232. doi: 10.1016/j.saa.2024.124232. Epub 2024 Mar 31.

Authors

Anand M Shrivastav¹, Nagham Ali², Neetika Singh³, Eitan Lunenfeld⁴, Ibrahim Abdulhalim⁵, Mahmoud Huleihel⁶

Affiliations

¹ Department of Electrooptics and Photonics Engineering, ECE School, Ilse-Kats Nanoscale Science and Technology Center, Ben Gurion University, Beer Sheva 84105, Israel; Department of Physics and Nanotechnology, College of Engineering and Technology, SRM Institute of Science and Technology, Kattankulthar, Tamil Nadu 603203, India.
² The Shraga Segal Dept. of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel; The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel.
³ Department of Electrooptics and Photonics Engineering, ECE School, Ilse-Kats Nanoscale Science and Technology Center, Ben Gurion University, Beer Sheva 84105, Israel.
⁴ Adelson School of Medicine, Ariel University, Ariel, Israel.
⁵ Department of Electrooptics and Photonics Engineering, ECE School, Ilse-Kats Nanoscale Science and Technology Center, Ben Gurion University, Beer Sheva 84105, Israel. Electronic address: abdulhlm@bgu.ac.il.
⁶ The Shraga Segal Dept. of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel; The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva 8410501, Israel. Electronic address: huleihel@bgu.ac.il.

PMID: 38593538
DOI: 10.1016/j.saa.2024.124232

Abstract

The present study aims to identify spermatogenesis in testicular seminiferous tubules (ST) and testicular tissue of adult normal and busulfan-treated mice utilizing PCA and Raman spectroscopy. Raman measurements were conducted on single tubules and testes samples from adult and immature mice, comparing them with those from busulfan-treated adult mice, with validation through histological examination. The analysis revealed a higher signal variability (30 %-40 % at the peaks), prompting scrutiny of individual Raman spectra as a means of spermatogenesis measurement. However, principal component analysis (PCA) demonstrated significant cluster separation between the ST of mature and immature mice. Similar investigations were performed to compare ST from normal mature mice and those from busulfan-treated (BS-treated) mature mice, revealing substantial separation along PC1 and PC2 for all comparison sets. Additionally, comparing testicular samples from mature and immature mice revealed distinct separation in PCA. The study concludes that the combined approach of PCA and Raman spectroscopy proves to be a noninvasive and potentially valuable method for identifying spermatogenesis in seminiferous tubules and testicular samples.

Keywords: Chemotherapy; Principal component analysis; Raman spectroscopy; Seminiferous tubules; Spermatogenesis.

MeSH terms

Animals
Busulfan*
Male
Mice
Principal Component Analysis*
Seminiferous Tubules* / drug effects
Spectrum Analysis, Raman* / methods
Spermatogenesis* / drug effects
Spermatogenesis* / physiology
Testis* / drug effects

Substances

Busulfan