H&E Staining in Rat Tissue

Haematoxylin and eosin staining in rat tissue to identify cellular structures.

Abstract

In order to identify cellular structures in our organs we used Haematoxylin & Eosin to stain the liver, kidney, muscle, spleen, lung, heart and intestine of a rat and compared similarities and differences within the structures of each organ. Haematoxylin is a naturally occurring chemical used as the basis of a dye in laboratories throughout the world to stain nuclei in microscope slide preparations. [1] Delafield’s Haematoxylin is an Aluminium haematoxylin which recommended for progressive staining of tissues. (i.e. staining for a predetermined time to adequately stain the nuclei but leave the background tissue relatively unstained, to be later counterstained.) [3] Eosin is the counterstained used to stain the cytoplasm of the tissue pink. Each slide was viewed under both high and low magnifcation to look for similarities and differences in each tissue. Some similarities such as arteries, veins, epithelium and muscle were all observed but the most importantly to identifying each of these organs is the differences and unique structures of each organ.

Method

Seven different organs were stained using the common Haematoxylin and eosin staining method. This was done in three separate groups, using two different stain times. The first method used stained the muscle and Kidney slides. The staining method started with 10 minutes in tap water this was followed by Staining the nuclei with Delafield’s Haematoxylin for 2 minutes. The slides were then rinsed in tap water for 3 minutes to rinse the excess stain. The slides were then counterstained with Eosin for 35sec coloring the cytoplasm of our slides pink. The slides were then rinsed in tap water for 1 minute, followed by being left in Ethanol (100%) for 2 minutes twice to dehydrate the tissue. The final step before mounting was 2 minutes in Histoclear. Between each step the slides were dabbed to help remove excess liquid....