histology
Histology analysis
There were five steps involved in histological preparation.
Step 1: Dissection and tissues collection
The testis tissues were cut with a sharp scalpel into slices of not more than 2-3 mm thick and 10 mm long. The fixation process was commenced immediately after removal of the tissue sample. The specimen tube was filled with 20 ml of 10% formalin. The tissue was placed inside the specimen tube.
Step 2: Dehydration
Dehydration process begins when the fixed tissues was placed in the specimen tube and immersed through a series of 10 ml alcohol solution with ascending concentration starting from 70% alcohol to 80%, 95%, 100% and another 100%, 1 hour for each step. When the dehydration proses was complete, two immersions of the tissue sample in xylen solution to clear out the alcohol.
Step 3: Infiltration and tissue embedding
For infiltration process, a small block of paraffin was placed in 500 ml beaker and was melted in the oven. The melted paraffin was poured into embedding mold and then, tissue sample was placed in paraffin wax filled mold. The base of tissue cassette was placed on top of the mold and was attached by adding further melted paraffin. The tissue specimen block was solidified at room temperature. The cassette that was filled with melted paraffin provided a stable base for clamping the rotary microtome.
Step 4: Samples sectioning and sample mounting
The paraffin section was cut at 5 μm of thickness with a rotary microtome. Sections of tissues obtained were floated on the surface of warm water in a floatation bath to flatten the sections. The sections were picked up onto the microscope slide, drained it in vertical position for several minutes. The slides were then placed on a slide holder in the oven at 37 °C overnight.
Step 5: Staining
The slides were placed in a staining rack and were dipped into xylene for 5 minutes and repeated once in new xylene for another 5 minutes. Then, the slides were dipped into
There were five steps involved in histological preparation.
Step 1: Dissection and tissues collection
The testis tissues were cut with a sharp scalpel into slices of not more than 2-3 mm thick and 10 mm long. The fixation process was commenced immediately after removal of the tissue sample. The specimen tube was filled with 20 ml of 10% formalin. The tissue was placed inside the specimen tube.
Step 2: Dehydration
Dehydration process begins when the fixed tissues was placed in the specimen tube and immersed through a series of 10 ml alcohol solution with ascending concentration starting from 70% alcohol to 80%, 95%, 100% and another 100%, 1 hour for each step. When the dehydration proses was complete, two immersions of the tissue sample in xylen solution to clear out the alcohol.
Step 3: Infiltration and tissue embedding
For infiltration process, a small block of paraffin was placed in 500 ml beaker and was melted in the oven. The melted paraffin was poured into embedding mold and then, tissue sample was placed in paraffin wax filled mold. The base of tissue cassette was placed on top of the mold and was attached by adding further melted paraffin. The tissue specimen block was solidified at room temperature. The cassette that was filled with melted paraffin provided a stable base for clamping the rotary microtome.
Step 4: Samples sectioning and sample mounting
The paraffin section was cut at 5 μm of thickness with a rotary microtome. Sections of tissues obtained were floated on the surface of warm water in a floatation bath to flatten the sections. The sections were picked up onto the microscope slide, drained it in vertical position for several minutes. The slides were then placed on a slide holder in the oven at 37 °C overnight.
Step 5: Staining
The slides were placed in a staining rack and were dipped into xylene for 5 minutes and repeated once in new xylene for another 5 minutes. Then, the slides were dipped into