Abstract

Manoj A.* and Annamma Paul*

ABSTRACT

The aim of the study was Fixation of tissues inorder to prevent Autolysis and Putrefaction, and Ensure the tissue processing was an adequate to get excellent paraffin sections, Identification of the Microscopic architecture of tissue by treating with Conventional staining and Calibration of the measurement of specimens. The Protocol of first day, samples were received and provide unique identification number, fixed in 10% Formaldehyde. On second day, tissue was transferred to the Automatic Tissue Processor (ATP) for undergoing Dehydration, Clearing, Infiltration and Impregnation with paraffin wax. On third day the tissue was embedded in paraffin wax in an L-block at 56°C. On fourth day the tissue containing paraffin block was subjected to section cutting by a Rotary Microtome. The tissue ribbon was flatten in a floatation bath and mounted on a microscope slide. The paraffin wax was de-waxed by hot plate (40° to 50°C) for 1 hour or dry overnight. The remaining wax was de-parffinized by keeping the slide in xylene for 5 minutes. Hydration of the tissue was undergone by descending grades of alcohol of 100%, 90%,80% and 70% for 30 to 60 second each. Staining was done by dipping the slide in Haematoxylin stain for 3 to 5 minutes and wash with running tap water for 15 seconds. Differentiation of nucleus and cytoplasm was done by dipping the slide in 0.5% hydrochloric alcohol. Slide was dipped in ammonia for further blueing. Slide was washed with 95% alcohol. Counter staining was undergone by treatment with Eosin-Y for 10-60 seconds. Dehydration was by washing the slide in ascending grades of alcohol 70%, 95% and 100%. Tissue was permanently mounted on slide by adding DPX and inverted on a cover slip. Microscopic architecture of tissue was identified by compound microscope with 10X and 40X magnifications. The actual size of the specimen was calibrated by Micrometry.