Luxol Fast Blue (LFB)

Validation Stain: Luxol Fast Blue (LFB) – Cresyl Violet
Other Applicable Stains: LFB, Bodian, Holzer, PTAH and Weil’s


  • Tissue sections are cut at 8 microns

(+) Spinal Cord
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4407A 10/set $53.00
Part # 4407B 98/set $372.30


Tissue:  Positive staining spinal cord.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 8 microns on Superfrost™ Plus slides.

Quality Control Stain:  Luxol Fast Blue quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.



With Luxol Fast Blue (LFB) Stain Set:  Part 12218A/B Individual Stain Solution   
Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic 500/1000 ml
Solution B: Lithium Carbonate, Saturated Aqueous 500/1000 ml Part 12215
Hematoxylin Stain, Harris Modified           Part 1201
Acid Alcohol 1% Part 10011



Newcomer Supply Luxol Fast Blue (LFB) Control Slides are for the positive histochemical staining of myelin sheath in central nervous system tissue and in peripheral nerve.

Periodic Acid Schiff (PAS), cresyl violet, hematoxylin or eosin stains/counterstains can be added to the LFB procedure for additionally enhanced staining.



  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.
    1. Stop at 95% ethyl alcohol; no distilled water rinse.
    2. See Procedure Notes #1 and #2.
  3. Incubate in Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic for 2 hours at 60°C or overnight at 37°C; seal lids tightly.

        Microwave Modification: See Procedure Note #3.

  1.   Place slides in a plastic Coplin jar (Part 5184) containing Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic and microwave at 70°C for 10 minutes.
  1. Rinse slides quickly in 95% ethyl alcohol; 2-3 dips.
  2. Rinse slides in distilled water.
  3. Prepare Working Lithium Carbonate 0.5%; combine and mix well.
    1. Solution B: Lithium Carbonate, Saturated Aqueous  20 ml
    2. Distilled Water                                                           20 ml
  4. Differentiate each slide individually; immerse slide in Working Lithium Carbonate 0.5%, Aqueous for 20-30 seconds.
    1. Save solution and reuse in Step #10a.
  5. Continue differentiation in 70% ethyl alcohol (Part 10844), until gray and white matter can be distinguished.  Do not over differentiate.
  6. Rinse slides in distilled water.
  7. Complete differentiation:
    1. Rinse briefly in Working Lithium Carbonate 0.5%, Aqueous.
    2. Rinse in two changes of 70% ethyl alcohol until greenish/blue white matter sharply contrasts with colorless gray matter.
  1. Rinse thoroughly in distilled water.
  2. Stain with Hematoxylin Stain, Harris Modified (Part 1201) for 1-5 minutes, depending on preference of stain intensity.
  3. Wash in running tap water for 3 minutes.
  4. Differentiate quickly in Acid Alcohol 1% (Part 10011); 3 dips.
  5. Wash well in running tap water.
  6. Blue in Solution B: Lithium Carbonate, Saturated Aqueous.
  7. Wash well in running tap water.
  8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.



Myelin Blue
Nissl substance and nuclei Blue
Other tissue components Dependent on other stains used



  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. The suggested microwave procedure has been tested at Newcomer Supply.  This procedure is a guideline and techniques should be developed for use in your laboratory
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps



  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 378.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 206-211.
  3. Klüver, Heinrich, and Elizabeth Barrera. “A Method for the Combined Staining of Cells and Fibers in the Nervous System.” Journal of Neuropathology and Experimental Neurology 12.4 (1953): 400-403.
  4. Modifications developed by Newcomer Supply Laboratory.