1.0 Purpose
To establish a standardized
procedure for iron staining of tissue sections and smears.
2.0
Scope
Applicable to all tissue sections
and smears received in the Surgical Pathology Cytopathology section for iron
staining.
3.0
Introduction
Potassium Ferrocyanide-HCL staining
method is used to detect iron in tissues and smears.
4.0
Principle
This reaction is considered the
first classical histochemical reaction. Treatment with an acid ferrocyanide
solution results in the marking of ferric iron in the form of hydroxide
(Fe(OH)) by dilute hydrochloric acid. The ferric iron then reacts with a dilute
Potassium Ferrocyanide solution to produce an insoluble blue compound, ferric
Ferrocyanide.
5.0
Analytical Measuring Range and Limit of Detection
Not Applicable.
6.0
Responsibility
- Technical staff
- Pathologist
7.0
Abbreviations
- HCL: Hydrochloric acid
8.0
Requirements
- Kit Reagents:
- 0.2% Aqueous Potassium Ferrocyanide
- 0.2% HCL
- 0.1% Eosin
- Xylene
- Absolute Alcohol
9.0
Precautions
- Handle all specimens with gloves and a mask.
- Handle all instruments with care to prevent injury.
10.0
Limitations & Interferences
Not Applicable.
11.0
Documentation
- Daily routine register
- Special stains log/register
12.0
Instructions
12.1
Pre Analytical Steps
- Sample preparation
- Reagent preparation
12.1.1
Working Solution
Mix equal volumes of 0.2% aqueous
Potassium Ferrocyanide and 0.2% HCL just before use.
12.2
Analytical Steps
- 12.2.1 Bring sections down to water by dipping in two
changes of xylene to remove the wax and two changes of alcohol to remove
xylene.
- 12.2.2 Wash in running tap water.
- 12.2.3 Treat with working solution for 30 minutes.
- 12.2.4 Wash in running tap water.
- 12.2.5 Counterstain with 0.1% eosin for 30 seconds.
- 12.2.6 Dehydrate, clear, mount, and label.
- 12.2.7 Submit the slides to the pathologist for
reporting.
12.3
Quality Control and Assurance
- 12.3.1 Known positive controls are run with each batch.
12.4
Calibration and Calibration Verification
Not Applicable.
12.5
Storage of Samples
- Slides are stored for 10 years.
13.0
Reference Range
Not Applicable.
14.0
Critical Value
Not Applicable.
15.0
Reporting of Results and Interpretation
Iron stains a Prussian blue color.
16.0
Contingency Plan
In case of reagent or manpower
shortage or instrument breakdown, suitably trained backup staff are available.
17.0
References
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the In-vivo Laboratory.
4. List of All SOPs and Documents for Clinical Research
5. List of Instruments and Equipment Used In Laboratory Animal Facility
6. Animal Facility Design - Small Laboratory Animals (Rat, Mice, Rabbit, Guinea Pig)
7. List of Chemicals Needed for In-vivo Laboratory
8. List of Physiological Data for Small Laboratory Animals
9. Leading Lab-Diet Suppliers in India and Beyond
10. List of All SOPs and Documents for Animal House Facility
11. List of SOPs and Documents for Histopathology Laboratory
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