METROPOLIS HEALTHCARE LTD

MAY - GRUNWALD GIEMSA STAIN FOR CYTOLOGY

SMEARS

STANDARD OPERATING PROCEDURE

SECTION SURGICAL PATHOLOGY

Standard Operating Procedure for May -

SOP NAME. Grunwald Giemsa stain for Cytology
smears
SOP NO. Surgical Pathology/Cytopath/SOP-22/V-I

******
SUPERSEDES

COPY NO. 01

Signature Date
PREPARED & ISSUED BY
Section Head May 10, 2010

APPROVED BY:
Head of the Department May 15, 2010

EFFECTIVE DATE May 17, 2010

Signature Date

REVIEWED ON & BY
Head of the Department

Metropolis Healthcare Ltd.

Surgical Pathology/Cytopath/SOP-22/V-I
Confidential- Do not reproduce or distribute 1 of 3
 METROPOLIS HEALTHCARE LTD
MAY - GRUNWALD GIEMSA STAIN FOR CYTOLOGY
SMEARS

Purpose of the Procedure: To establish the Standard Operating Procedure

for May - Grunwald Giemsa stain for Cytology smears

Principle: May - Grunwald Giemsa stain is Romanowsky staining technique

containing two dyes Methylene blue and eosin Y.
Methylene blue on oxidation produces coloured compound known as Azure
that has ability to combine with eosin.
Methylene blue azure is blue violet and stains mainly acidic cell components
where as eosin is red in colour and stains more basic structures where as
Giemsa is traditional nuclear stain.

Preparation of solution:

1) May - Grunwald stock

May-Grunwald powder – 2.5gm

Methanol - 1000ml
Mix and store at room temperature in a well stoppered bottle.
May-Grunwald working solution:
Stock solution 2ml
Methanol 8ml
To be prepared fresh everyday

2) Giemsa stock solution.

Giemsa powder - 1gm

Glycerin - 54ml.
Methanol - 84ml

3) Stock Buffer Solution (pH 6.8)

K2HPO4 - 2.7gm% in distilled water.

NaOH - 0.8gm% in distilled water.
Mix 50ml of K2HPO4 with 236 ml of NaOH
Working Buffer Solution:
Take 40ml of stock buffer solution and make up to 1000ml.

4) Giemsa working Solution.

It is prepared fresh everyday by mixing 1ml of Giemsa solution and

9ml working buffer solution.

Procedure:
Metropolis Healthcare Ltd.
Surgical Pathology/Cytopath/SOP-22/V-I
Confidential- Do not reproduce or distribute 2 of 3
 METROPOLIS HEALTHCARE LTD
MAY - GRUNWALD GIEMSA STAIN FOR CYTOLOGY
SMEARS

1) Fix smears in methanol for 15 mins.

2) Stain the smears with working May-Grunwald solution for 25 mins.
3) Wash in tap water
4) Stain the smears with working Giemsa stain for 20 mins.
5) Wash in running tap water, air dry & Mount with DPX

Metropolis Healthcare Ltd.

Surgical Pathology/Cytopath/SOP-22/V-I
Confidential- Do not reproduce or distribute 3 of 3