Im using a fast method (-+ 4 hours) for clearing and dehidration before embedding. Then i left the testis on the wax for 1 day before embedding. The fixation that i use is 10% formalin. Help me please, im new to histotechnique. Is it on the microtome or its because of the way i do the tissue before wax section?
Hi, it's likely that the testis is not processed properly. You can use this method which i have used for rat stomachs and testes.
Also, the fixation with formalin has to be 24 hours before tissue processing.
Before you section the block, put it in -20 freezer for half an hour so that it is harden, then the sectioning will be easier. Hope it helps!
It is no miracle that your samples 'break' and/or crumble on sectioning... "Rapid or Fast methods" in Histology for delicate, but dense 'capsular' wrapped tissues/organs like testes/testis always need longer time for processing (but perhaps ou've minimzed specimen size /volume we do not know about.
It would be of benefit for answering more precisely if yyou, Arief Sirajuddin , would include a protocol sheet with your Question (you could do that easily also in the Question by editing, or just answer and attaching a pdf or word.docx-file).
NB:
Chapter Preparation of Testicular Samples for Histology and Immunohi...
For more details on importance of thourough preparation /fixation of testis cf. also: Article Advanced immunostaining approaches to study early male germ ...
(=NOTE the two undersign characters have to be removed in your browser to get the access to the pdf /free ACCESS & Download: >https__://www.sciencedirect.com/science/article/pii/S1873506118300370
Si usted ha seguido un protocolo parecido al de Faizatul debe tener también presente el tiempo de permanencia en cada una de los reactivos usados. Un tiempo muy corto en ellos no permitirá una buena deshidratación o aclaramiento y por tanto no habrá un buen embebido de parafina. Un exceso de tiempo en ellos volverá los tejidos quebradizos, sobre todo la permanencia excesiva en parafina, o si la parafina está a una temperatura muy superior a la recomendada.
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