Histotechnology Technical methods

Stain for Helicobacter pylori

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GIMENEZ STAIN

SECTIONS

3-4 um paraffin


METHOD

1. Sections to water.

2. Filter on carbol fuchsin solution for 1 minute.

3. Wash well in water.

4. Stain with 1% malachite green for up to 45 seconds.

5. Wash well in water.

6. Repeat step 4 until sections appear blue/green.

7. Wash, blot dry, and dry in air.

8. Mount sections in DPX


RESULTS

--Helicobacter red/magenta

--Background blue/green


SOLUTIONS

Malachite Green

--Malachite green 0.8g

--Distilled water 10mml

Stock carbol fuchsin solution

--Basic Fuchsin 1g

--Absolure alcohol 10ml

--5% aqueous phenol 10ml

--Filter before use

Buffer solution (Phosphate buffer pH7.5 0.1M)

--0.1M Sodium di-hydrogen orthophosphate 3.5 mls

--0.1M Di-sodium hydrogen orthophosphate 15.5 mls

Working solution (Filter onto sections for use)

--Phosphate Buffer 10ml

--Stock Carbol fuchsin Solution 4 mls


SAFETY Carbol fuchsin contains phenol.

NOTE: No responsibility is assumed by The University of Nottingham or the Queens Medical Centre NHS Trust for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions or ideas contained in the material herein. It is the users responsibilty to ensure that all procedures are carried out according to appropriate Health and Safety requirements.


© Copyright 1997 University of Nottingham Medical School Division of Histopathology. This page was last built on Mon, May 5, 1997 with Frontier. Thanks for looking in. Comments to James.Lowe@nottingham.ac.uk