title: Different fixative methods influence histological morphology and TUNEL staining in mouse testes.
abstract: Society of Toxicologic Pathology has recommended mDF to fix testes since 2002. However, subsequent studies showed that false TUNEL-positive cells were observed in mDF-fixed testes. This study compared the effects of different fixation methods on histology and TUNEL staining in mouse testes. Results showed that fixation for 24 or 36h in mDF provided better morphologic details in untreated testes, but markedly enhanced false TUNEL-positive staining. To optimize the fixation, testes were fixed using mDF for 6h and then PFA for 18h. Interestingly, fixation using mDF/PFA manifested better morphologic details, and rarely caused false TUNEL-positive cells in testes. Finally, we examined germ cell apoptosis in testes using mDF/PFA fixation in cadmium-treated mice. As expected, cadmium triggered germ cell apoptosis which was well visualized in the mDF/PFA fixed testes. Taken together, mDF plus PFA fixation not only minimizes false TUNEL-positive cells, but also provides integrated morphologic details in testes.
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China. Electronic address: wanghuadev@126.com.
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China.
authors.forename: Lu-Lu
authors.initials: LL
authors.lastname: Yang
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China.
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Ji
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China; Department of Histology and Embryology, School of Basic Medicine, Anhui Medical University, Hefei, Anhui, China.
authors.forename: Yuan-Hua
authors.initials: YH
authors.lastname: Chen
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Hu
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Toxicology, School of Public Health, Anhui Medical University, Hefei, Anhui, China. Electronic address: xudex@126.com.
authors.forename: De-Xiang
authors.initials: DX
authors.lastname: Xu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002104:Cadmium / Q000633:toxicity; D005404:Fixatives; D005557:Formaldehyde; D020287:In Situ Nick-End Labeling / Q000379:methods; D008297:Male; D051379:Mice; D011108:Polymers; D013194:Staining and Labeling; D013737:Testis / Q000033:anatomy & histology* / Q000166:cytology / Q000187:drug effects; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D005404:Fixatives; D011108:Polymers; D002104:Cadmium; D005557:Formaldehyde; C003043:paraform
keywords: Modified Davidson’s; Mouse; Paraformaldehyde (PFA); TUNEL staining; Testis; fluid (mDF)