RESUMO
Histopathological examination of testes is important in assessing spermatogenesis and testicular function. Modified Davidson's fluid (mDF) has been proposed as a superior substitute for Bouin's fluid (BF) for fixation of adult animal testes. Besides, 4% paraformaldehyde (PFA) has been commonly used to fix testes with convenience. We compared the morphology of the rat testis fixed in 4% PFA, mDF, or BF using hematoxylin and eosin (HE)-stained sections. Fixation in 4% PFA resulted in obvious tissue shrinkage artifacts, especially between seminiferous epithelium cells. Shrinkage artifacts were also observed in the central area of the testes fixed in BF. Use of mDF did not cause shrinkage artifacts between seminiferous tubules, though a small amount can be observed in seminiferous tubules between germ cells. Clarity of nuclear detail in testes fixed in mDF and BF is better compared to 4% PFA. Our study demonstrated that fixation in mDF provided better morphologic details in the rat testis as compared with 4% PFA and BF.
Assuntos
Fixadores/classificação , Testículo/anatomia & histologia , Fixação de Tecidos/métodos , Animais , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Fourier transform infrared microspectroscopy (FTIRM) and infrared imaging (FTIRI) are techniques utilized in the analysis of bone mineral and matrix properties in health and disease. Since the spatial arrangement of bone tissue is conserved using FTIRM and FTIRI, quantitative data can be obtained on bone mineral (hydroxyapatite) crystalline size and composition, and on matrix structure and composition at discrete anatomic locations with a spatial resolution from approximately 7 mm (FTIRI) to 10 mm (FTIRM). To section bone for FTIRM and FTIRI, it must be preserved ("fixed") to maintain its properties, and embedded in a hard supportive material. Since most of the embedding media have components that spectrally overlap the components of mineralized tissues, it is critical to define optimal embedding and fixation protocols that have the least effect on mineral and matrix spectra. In the current study, the spectra of mouse calvaria in seven different fixatives and six different commonly used embedding media were assessed by FTIRM and FTIRI. The fixatives evaluated were absolute ethanol, 70% ethanol, glycerol, formaldehyde, EM fixative, and formalin in cacodylate or phosphate-buffered saline. The embedding media tested were Araldite, Epon, JB-4, LR White, PMMA, and Spurr. Comparisons were made to FTIR spectra obtained from unprocessed ground calvaria and to spectra of cryosections of unfixed tissue, fast-frozen in polyvinyl alcohol (5% PVA). Non-aqueous fixatives and embedding in LR White, Spurr, Araldite, and PMMA had the least effect on the spectral parameters measured (mineral to matrix ratio, mineral crystallinity, and collagen maturity) compared with cryo-sectioned calvaria and non-fixed, non-embedded calvaria in KBr pellets.
Assuntos
Osso e Ossos , Calcificação Fisiológica , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Inclusão do Tecido/métodos , Fixação de Tecidos/métodos , Animais , Densidade Óssea/fisiologia , Osso e Ossos/citologia , Osso e Ossos/metabolismo , Fixadores/classificação , Camundongos , Crânio/anatomia & histologia , Crânio/metabolismoRESUMO
Técnica histológica: pasos, artifisios de técnicas, técnicas histoquímicas y inmunohistoquímicas. Microscopía: fundamentos de la óptica física y geométrica; lentes: definición, clasificación, elementos; microscopio fotónico; tipos especiales de microscopios; unidades de medidas usadas en microscopía
Assuntos
Histologia , Técnicas Histológicas/instrumentação , Imuno-Histoquímica/métodos , Imunofluorescência/instrumentação , Técnicas Imunoenzimáticas/instrumentação , Histocitoquímica , Fixadores/classificação , Fixadores/normas , Fixadores , /instrumentação , /métodos , /normas , Microscopia de Contraste de Fase/instrumentação , Microscopia de Interferência/instrumentação , Microscopia de Polarização/instrumentação , Microscopia de Fluorescência/instrumentação , Microscopia Ultravioleta/instrumentação , Microscopia Eletrônica de Varredura/instrumentação , Microscopia Eletrônica/instrumentação , Microscopia/instrumentação , Microscopia/métodos , Óptica e Fotônica/instrumentação , Lentes/normas , Padrões de ReferênciaRESUMO
Técnica histológica: pasos, artifisios de técnicas, técnicas histoquímicas y inmunohistoquímicas. Microscopía: fundamentos de la óptica física y geométrica; lentes: definición, clasificación, elementos; microscopio fotónico; tipos especiales de microscopios; unidades de medidas usadas en microscopía
