Recent advancements of flow cytometry: new applications in hematology and oncology

Abstract

Flow cytometry offers great diagnostic opportunities in the vast majority of hematologic and oncologic diseases with multiple cellular and molecular information within an individual cell. We will discuss various applications of flow cytometry, particularly in hematology and oncology, in addition to general principles and limitations of flow cytometry. They include nucleic acid analyses in cancer cells, new methods for assessing rare circulating tumor cells and disease-specific applications in malignancy with emphasis on diagnosis and treatment of hematologic malignancy, including minimal residual disease. With improvement of monoclonal antibodies, fluorescence and laser technology, flow cytometry now offers new avenues of assessing cellular functionality through examination of intracellular compartments. High-throughput quantitative analysis, advancements of in vivo flow cytometry and assessment of minimal residual diseases, as exampled in patient stratification and prediction of leukemia therapeutic response, will further make flow cytometry indispensable in medicine.

Keywords::

• applications
• flow cytometry
• in vivo flow cytometry and standardization of flow cytometry
• minimal residual disease
• qualitative and functional assays

Financial & competing interests disclosure

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending or royalties.

No writing assistance was utilized in the production of this manuscript.

Key issues

• Flow cytometry is a powerful tool with the capacity to analyze thousands of single cells within seconds, while obtaining multiple cellular parameters that may yield important diagnostic information.

• The specific immunophenotypic signature of malignant cells, as established at diagnosis and during the course of treatment, is used for evaluation of therapeutic response after treatment and detection of minimal residual disease.

• Quantification of minimal residual leukemic cells can estimate the reduction of disease burden following induction therapy and help predict disease outcome. Recently, the European LeukemiaNet and the EuroFlow Consortium have established standardized protocols and software tools to analyze immunophenotypes of normal and malignant leukocytes in bone marrow and blood, employing eight-color flow cytometry.

• New advances in multiphoton cytometry, including photoacoustic and photothermal methods, in combination with enhanced contrast nanoparticles enabled detection of rare circulating tumor cells in blood or lymphatic vessels before metastasis or tumor formation occurs in vivo.

• Flow cytometry can assay intracellular antigens such as DNA, cytokines and phosphorylated signaling proteins, which allows functional analysis and helps establish therapeutic strategies to stratify high-risk cancers and predict therapeutic response.

Notes

AML: Acute myeloid leukemia.