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Evaluation of immunocytochemistry on destained giemsa stained smears as an alternative to conventional technique
Pritika Kushwaha1, Toshali Pandey1, Radhika Agarwal1
1Department of Pathology, Maulana Azad Medical College, New Delhi, India.
Insights
This study found that de-staining May-Grünwald Giemsa (MGG)-stained smears for immunocytochemistry (ICC) is not a reliable alternative to standard ICC. The protocol yielded unsatisfactory results for both pancytokeratin and leukocyte common antigen (LCA) staining.
Area of Science:
- Pathology
- Cytology
- Immunohistochemistry
Background:
- Establishing immunocytochemical (ICC) staining protocols for May-Grünwald Giemsa (MGG)-stained smears presents challenges.
- Utilizing pre-stained slides for ICC is desirable for timely diagnoses and patient convenience.
Purpose of the Study:
- To evaluate and compare the efficacy of MGG-stained smears for ICC after de-staining and rehydration against the standard ICC technique.
Main Methods:
- A prospective study analyzed 40 Fine Needle Aspiration Cytology (FNAC) samples.
- Smears were stained with MGG, then de-stained and subjected to ICC for pancytokeratin or leukocyte common antigen (LCA)/CD45.
- Staining results were semi-quantitatively graded based on intensity, localization, uniformity, counter-staining, and background.
Main Results:
- The de-staining protocol was inferior to conventional ICC across all evaluated parameters.
- Optimal staining intensity for pancytokeratin was achieved in only 8% of de-stained smears, compared to 44% for wet-fixed smears.
- Optimal staining for LCA was achieved in 14.3% of de-stained smears versus 85.7% for wet-fixed smears.
Conclusions:
- The proposed protocol of using de-stained MGG smears for ICC is not a successful alternative to the conventional technique.
- This method failed to produce satisfactory results for diagnostic ICC staining.
Introduction:
Protocol for immunocytochemical (ICC) staining in May-Grünwald Giemsa (MGG)-stained smears has been difficult to establish. It is the need of the hour to be able to use prestained slides for ICC in specific cases to deliver timely diagnoses and reduce inconvenience to patients.
Aims And Objectives:
To evaluate and compare the use of MGG-stained smears for the purpose of ICC, after de-staining and saline rehydration to that of routine standard ICC.
Materials And Methods:
A prospective study was conducted on 40 FNAC samples: 25 cases of breast disease and 15 cases of reactive lymphoid hyperplasia known to express pancytokeratin and leukocyte common antigen (LCA)/CD45, respectively. Air-dried smears of each case were stained by standard MGG stain and after the report was dispatched, one smear was selected and sent for ICC. The smears were analyzed to determine the overall result and grade each smear semi-quantitatively with respect to staining-intensity, stain-localization, staining-uniformity, counter-staining, and background-staining.
Observations And Results:
The proposed protocol was inferior to conventional ICC in all the parameters, more pronounced in pancytokeratin than LCA/CD45. Only 8% of air-dried smears stained for pancytokeratin showed optimal stain intensity (as opposed to 44% of wet-fixed smears), whereas only 14.3% of air-dried smears were optimally stained for LCA (as opposed to 85.7% of wet-fixed smears).
Conclusion:
The proposed protocol of de-stained Giemsa smears as an alternative to conventional technique for ICC was unsuccessful in giving satisfactory results.
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