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Electron Microscopy Sciences

Technical Data Sheets

Fat Stains In Supersaturated Alcohol

EMS Catalog #: 26503-Series

Fixation:

Formalin.

Sections:

Frozen

Preparing Working Stain

Dilute 6 ml 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 Hematoxylin or Ehrlich’s Hematoxylin 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

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Fat Stains In SuperSaturated Alcohol