Brief Report - (2025) Volume 10, Issue 2
Received: 03-Mar-2025, Manuscript No. jmhmp-25-168570;
Editor assigned: 05-Mar-2025, Pre QC No. P-168570;
Reviewed: 17-Mar-2025, QC No. Q-168570;
Revised: 24-Mar-2025, Manuscript No. R-168570;
Published:
31-Mar-2025
, DOI: 10.37421/2684-494X.2025.10.278
Citation: Gaur, Karen. “A Comparison of IHC and Flow Cytometry in Detecting Hematologic Malignancies.” J Mol Hist Med Phys 10 (2025): 278.
Copyright: © 2025 Gaur K. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.
This is particularly valuable in cases involving bone marrow biopsies or lymph node excisions, where architectural features such as follicular patterns, sinusoidal involvement, or interstitial infiltrates are diagnostically relevant. IHC is indispensable in the diagnosis of lymphomas, especially when evaluating paraffin-embedded tissue and is widely used for classifying subtypes according to the WHO criteria. Antibodies against markers such as CD3, CD20, CD30, BCL2 and Ki-67 help characterize T-cell and B-cell populations, detect proliferative indices and determine clonality. Flow cytometry, in contrast, is a quantitative technique that analyzes the physical and antigenic properties of individual cells in suspension. Using fluorescent-labeled antibodies, FCM can rapidly analyze thousands of cells per second, producing high-dimensional data that offers detailed immunophenotypic profiles. This approach is highly sensitive and capable of detecting Minimal Residual Disease (MRD), making it invaluable in the diagnosis and follow-up of leukemias and lymphoproliferative disorders. FCM excels in detecting abnormal populations based on aberrant expression of lineage-specific markers, maturation antigens, or combinations not normally seen in healthy hematopoiesis. Commonly analyzed markers include CD19, CD45, CD10, CD34, HLA-DR and lineage-specific antigens such as MPO and TdT [3].
While IHC offers the advantage of contextual histological information, it is generally more limited in terms of multiplexing capacity compared to flow cytometry. Typically, only a small number of markers can be evaluated per tissue section and analysis is often semi-quantitative. In contrast, modern multicolor flow cytometry panels can simultaneously assess 8 to 20 or more antigens on a single cell, allowing for more refined classification and the detection of subtle immunophenotypic changes indicative of malignancy. Moreover, FCM is generally faster, with results often available within hours, whereas IHC may require longer processing times, especially in formalin-fixed paraffin-embedded specimens. Despite these differences, the two methods are complementary rather than mutually exclusive. In the diagnosis of acute leukemia, for example, flow cytometry is frequently used for initial classification and to distinguish between myeloid and lymphoid lineages. IHC then plays a confirmatory role, especially when assessing the tissue context or evaluating myeloid sarcomas and lymphoblastic infiltrates in solid organs. In the workup of lymphomas, IHC is essential for identifying histological subtypes and detecting markers such as BCL6, CD23, or cyclin D1, while flow cytometry is often employed for peripheral blood or bone marrow evaluation to detect disseminated disease or clonality [4].
Both techniques also present challenges. Flow cytometry requires viable cells and thus cannot be performed on fixed tissue, limiting its use in archival samples. Cell viability, sample handling and cellular yield are crucial for obtaining reliable FCM data. IHC, meanwhile, may suffer from antigen degradation due to fixation and variability in staining protocols or antibody specificity can affect interpretation. Furthermore, subjective interpretation of staining patterns in IHC may lead to interobserver variability, whereas FCM offers more objective numerical data, albeit requiring expertise in gating strategies and interpretation. Advances in technology are increasingly blurring the boundaries between these two approaches. Multiplex IHC and immunofluorescence techniques are expanding the capabilities of tissue-based analysis, allowing multiple antigens to be assessed in the same section. Similarly, mass cytometry and spectral flow cytometry offer even higher dimensionality in cell analysis, potentially revealing novel diagnostic and prognostic biomarkers. Integration of these modalities with molecular techniques such as Next-Generation Sequencing (NGS), Fluorescence In Situ Hybridization (FISH) and digital pathology platforms is paving the way for more precise and comprehensive hematologic cancer diagnostics [5].
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