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