USA - (800) 526 3867
UK - 44 (0) 1482 646020
Staining Procedures

Most dyes used to visualize the membranes and organelles of the cell are water soluble. The embedded wax must therefore be removed prior to staining. This is done by effectively reversing the tissue processing schedule.
There are literally thousands of staining protocols and procedures in use. As an example, one of the most common stains, the Hematoxylin-Eosin stain, is presented below. For a detailed list of stain procedures we recommend that you visit the University of Bristol web site.
Hematoxylin and Eosin Stain:
Hematoxylin is a natural compound extracted from a species of tree found in Mexico and the West Indies. The extracted compound is then oxidized to produce hematein, which is the active staining component of the hematoxylin stain. Hematoxylin stains must therefore be 'ripened' by oxidation before they can be used. Hematoxylin staining requires the use of a mordant (most commonly aluminum salts) and stains the nuclear components of cells a dark blue. Hematoxylin is used in combination with eosin because eosin stains the cytoplasmic organelles varying shades of pink, red or orange. The combination of the two stains provides a broad range of morphological information about the section.

H & E Stain with Ehrlich's Hematoxylin

FORMULATE HEMATOXYLIN AND EOSIN SOLUTIONS
Erlich's Hematoxylin: 2g hematoxylin
100ml ethanol
100ml glycerol
100ml deionized water
10ml glacial acetic acid
15g potassium alum
Dissolve the dye in the ethanol. Add all other components and allow to ripen for 2 months in direct sunlight, or ripen immediately with 100mg of Sodium Iodate. Note that chemical ripening will shorten the shelf life of the product considerably.
Eosin: 1g Eosin Y
100ml deionized water
STAINING PROCEDURE
  1. Stain rehydrated sections in Hematoxylin solution for 20-40 minutes.

  2. Wash in tap water for 1-5 minutes, until sections turn blue (ÒbluingÓ)


  3. Differentiate sections in 70% ethanol containing 1% HCl, for 5 seconds. This removes excess dye, allowing nuclear details to emerge.


  4. Wash 1-5 minutes in tap water, until blue.


  5. Stain in Eosin Solution 10 minutes.


  6. Wash 1-5 minutes in tap water.


  7. Dehydrate, clear and mount.

Note the use of tap water in the washing steps - tap water provides the alkanlinity necessary for the "bluing" process.




0

REQUEST A CATALOG

CURRENCIES

NEW PRODUCTS

UreaGel 29:1 Denaturing Gel System

Allows researchers to easily and safely produce denatururing gels of 29:1 acrylamide: bisacrylamide formula in concentrates up to 20% monomer.
Read more...


ProtoGel Sample Prep Kit

An easy-to-use, inexpensive kit that allows for pure samples and more concentrated SDS-PAGE samples.
Read more...


UreaGel Loading Buffer

Denaturing loading buffer for UreaGel as well as other denaturing polyacrylamide gel applications
Read more...

































Copyright © 2010 National Diagnostics. All rights reserved.
Please read our policies on privacy and shipping & returns.  Contact Us.
Powered by OS Commerce