Romanowsky Stains - Practical Task

Arlo Guthrie — Mon, 20 Apr 2009 13:00:13 GMT

Current Revision posted to Revision Guide For Student Nurses - Part 2 by Arlo Guthrie on 4/20/2009 1:00:13 PM

INTRODUCTION
The Romanowsky stains are used to stain smears in order to provide better cellular definition and also identify blood parasites such as haemobartinella.
They use 2 dyes:

HAEMATOXYLIN - which stains basic tissue blue.
EOSIN - which stains acidic tissue red.

There are 3 main types:

DIFF QUIK - very quick.
GIEMSA - ideal for staining haemobartinella.
LEISHMAN'S - excellent cellular definition.

EQUIPMENT

Lab coat and protective gloves.
Sink.
Drainer (for Giemsa and Leishman's).
Slide rack.
Paper towel to protect work surfaces from the stains.
Pipettes (for Giemsa and Leishman's).
Buffered distilled water.
Alcohol/methanol (for Giemsa).
Stains.
Pencil for labelling smears.
Watch or timer

METHOD

Put on lab coat and protective gloves; ensure that long hair is tied back and any jewellery removed.
Label the smears in pencil (if this has not already been done); information should include the date, owner's surname and animal identification.
Ensure that you have a flat, clean, dry surface upon which to work.
Handle the slides correctly; ie at the edges only.

Diff Quik.

Place the slide in the fixer (pale blue solution) for 5 seconds.
Place the slide in the red solution for 30 seconds.
Place the slide in the dark blue solution for 10 seconds.
Rinse gently with distilled water.
Air dry and place on a slide rack.

Giemsa

Fix slide in alcohol/methanol for 3 seconds.
Place the slide the correct way up on a drainer over the sink.
Use a pipette to flood the slide with Giemsa stain.
Leave for 30 minutes (if this task is given to you in the examination - you will be told to pretend that the time is up).
Rinse gently with distilled water.
Air dry and place on a slide rack.

Leishman's.

Place the slide the correct way up on a drainer over the sink.
Use a pipette to flood the slide with Leishman's stain.
Allow to stand for 15 minutes until a metallic scum appears on the solution surface (again, if this is an examination task - you will have to pretend that the 15 minutes is up).
Rinse gently with buffered distilled water.
Air dry and place on a slide rack.

EXAMINATION

Blood smears are examined under oil immersion (high power).

NB. Please refer to Module 4 - Blood Cells for further information.