EMS26503


Components:

Sudan IV-Isopropanol Solution

Oil Red O-Isopropanol Solution

Sudan III-Isopropanol Solution

Mayer's Haematoxylin

Erhlich’s Haematoxylin, Diluted

Sodium Phosphate 1%  Aqueous


Fixation:

Formalin


Sections:

Frozen


Preparing Working Stain

Dilute 6ml of stock solution Sudan IV in Isopropanol or Oil Red O in Isopropanol or Sudan III Isopropanol solution with 4 ml of distilled water. Allow to stand for 5-10 minutes and then filter. The filtrate may be used for several hours


Procedure

  1. Stain thin frozen sections with the working fat stain for 10 minutes. Wash in tap water.
  2. Stain for 5 minutes in either Mayer’s Haematoxylin or Ehrlich’s Haematoxylin diluted.
  3. Place in Sodium Phosphate 1 %, or in tap water until blue.
  4. Float out in water, take up on a slide and mount on Apathy’s Gum Syrup or Zwemer’s Glychrolgel or any other soluble aqueous medium.


Stain Results:  

Nuclei

Blue

Fat

Orange-Red

Erthrocytes

Sometimes Green

Cytoplasm

Lighter Green


References:

Clark, G, ed.: Staining Procedures, 3rd ed.: Baltimore: Williams & Wilkins Co., 1973, p. 149