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Updated: May 20, 2026

Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
High agreement of routine cytopathology and immunocytochemistry in canine lymphomas
R Sapierzyński1, I Dolka, M Fabisiak
1Department of Pathology and Veterinary Diagnostic, Faculty of Veterinary Medicine, Warsaw University of Life Sciences (SGGW), Nowoursynowska 159c, 02-766 Warsaw, Poland. rafal_sapierzynski@sggw.pl
Insights
Routine cytopathology and immunocytochemistry (IC) show high concordance (90%) for canine lymphoma immunophenotyping. This combination aids precise diagnosis, treatment planning, and prognosis, reducing the need for invasive biopsies.
Area of Science:
- Veterinary Pathology
- Immunology
- Oncology
Background:
- Accurate immunophenotyping is crucial for diagnosing canine lymphoma.
- Routine cytopathology using Giemsa stain is a common diagnostic method.
- Immunocytochemistry (IC) provides specific antigen detection for cell lineage confirmation.
Purpose of the Study:
- To compare the concordance between routine cytopathology (Giemsa stain) and immunocytochemistry (CD3, CD79 alpha) for canine lymphoma immunophenotyping.
- To evaluate the diagnostic accuracy of combining these methods.
- To assess the clinical utility of this combined approach in canine lymphoma cases.
Main Methods:
- Cytological samples from 70 canine lymphomas were analyzed.
- Samples were stained with Giemsa solution for routine cytopathology.
- Immunocytochemistry was performed to detect CD3 and CD79 alpha antigens.
- Cohen's kappa coefficient was calculated to measure concordance.
Main Results:
- Immunocytochemistry classified 42 lymphomas as B cell and 28 as T cell.
- Full accordance between cytopathology and IC was achieved in 63 out of 70 cases (90%).
- The combined approach yielded precise diagnoses in 90% of cases.
Conclusions:
- Routine cytopathology with Giemsa stain is a valuable tool for preliminary lymphoma immunophenotyping in dogs.
- Combining routine cytopathology with immunocytochemistry offers high diagnostic accuracy (90%) for canine lymphomas.
- This integrated diagnostic strategy simplifies treatment planning and prognosis determination while minimizing invasive procedures and associated costs.
Abstract:
The aim of this study was to compare the concordance of immunophenotype established with routine cytopathology (slides stained with Giemsa solution) and immunocytochemistry according to CD3 and CD79 alpha immunoreactivity. The study was performed on cytological samples of 70 canine lymphomas subtyped on the basis of the updated Kiel classification system. Additionally, cytologic samples were examined immunocytochemically for the CD3 and CD79 alpha antigens presence and thus immunophenotype of neoplastic growth was confirmed. The cytopathological and immunocytochemical diagnoses were then compared; in order to measure the concordance between immunocytochemistry (IC) result and Giemsa stain result of the same sample the Cohen's kappa coefficient was calculated. On the basis of the results of immunocytochemistry of 70 cases of canine lymphoma examined 42 were recognized as B cell lymphoma and 28 as T cell lymphoma. Full accordance between the results of routine cytopathology and IC was obtained in 63 out of 70 examined dogs (90% of cases). It can be concluded that cytopathological examination of Giemsa stained smears is helpful in determining the lymphoma cells immunophenotype. Additionally, it seems that combination of routine cytopathology and immunocytochemistry in cases of canine lymphomas allows to obtain the precise diagnosis in 90% of cases, and allows to receive most important information that is necessary for planning of appropriate therapy and to determine prognosis. Finally, this routine procedure allowed to eliminate the need of collection of tissue samples during surgery or core-biopsy and thus time, cost and patient discomfort related to more complex and invasive medical procedures can be easily reduced.
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