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Staining techniques: Leishman's stain

ISSN 2398-2977


Podcast: Staining techniques: Leishman's stain

Introduction

Rationale
  • A neutral stain for blood smears devised by the British surgeon, W B Leishman (1865-1926).
  • Double staining compound of Methylene blue and Eosin in alcohol, which is usually diluted and buffered before use.
  • It stains the different components of blood in a range of shades between red and blue.

Uses

  • Stain WBC, RBC and platelets to identify pathology.

Cons

  • Referral laboratory or human medical institution.

Alternatives

Work time

  • Not applicable - referral laboratory.

Requirements

People competence
  • Veterinary: medium.
  • Nursing: medium.

Method

  • Prepare blood smear   Blood smear  or cytospin slide.
  • Fix the wet preparation either by spraying it with a smear fix or gently dipping the slide in a Coplin jar of acetic acid.
  • Air dry for blood smear.
  • Place slide on a rack and flood with Leishman stock: 2 min for blood; 30 sec to 1 min for fluid.
  • Add 2 ml of pH 6.8 Sorensen's buffer and gently rock the rack to mix the solutions, then leave for 7 min; ~2 min for fluid.
  • Rinse in distilled water.
  • Air dry or carefully dry with fiber-free blotting paper.
  • Mount slide.
  • Examine under light microscope.

Further Reading

Publications

Refereed papers

Other sources of information

  • Dacie J V Sir & Lewis S M (2001) Practical Hematology. 9th edn. W B Saunders Co. ISBN 0-4430-6377 X.
  • Bain B J (1989) Blood Cells - A Practical Guide. Gower Medical Publishing. pp 8-10.
  • Hall R & Malia R (1984) Medical Laboratory Hematology. Butterworth & Co Ltd. pp 132-136.

Organization(s)

  • Howard Hughes Medical Institute, University of California, Los Angeles, 5-748 MacDonald Research Laboratories, 675 Charles E Young Drive South, Box 951662, USA. Website: www.hhmi.org.
  • University of Bristol, Department of Pathology and Microbiology, School of Medical Sciences, University Walk, Bristol BS8 1TD, UK. Website: www.bris.ac.uk/Depts/PathAndMicro/CPL/leishman.html.