Immunophenotyping
Immunophenotyping - Related Videos
08:17
A Semi-automated Approach to Preparing Antibody Cocktails for Immunophenotypic Analysis of Human Peripheral Blood
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Cited by 9 •
2016
Whole blood Immunophenotyping is indispensable for monitoring the human immune system. However, the number of reagents required and the instability of specific fluorochromes in premixed cocktails necessitates daily reagent preparation. Here we show that semi-automated preparation of staining antibody cocktail helps establish reliable immunophenotyping by minimizing variability in reagent dispensing.
09:12
Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
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Cited by 17 •
2014
One key to successful investigation of microglial biology is the preservation of microglial immunofunction ex vivo during isolation from CNS tissue. Isolating microglia via rotary shaking results in highly pure and immunofunctional cell cultures as assessed by fluorescent imaging, immunocytochemistry, and ELISA following microglia activation with the proinflammatory stimuli lipopolysaccharide (LPS) and Pam3CSK4 (Pam).
12:36
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
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Cited by 7 •
2018
Here we describe a single-cell proteomic approach to evaluate immune phenotypic and functional (intracellular cytokine induction) alterations in peripheral whole blood samples, analyzed via mass cytometry.
05:34
Operating and Biocontainment Procedures of a Facility for Laboratory Mice with a Natural Microbiome: Immunophenotyping Procedure
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2024
Here, we describe the structure and operating procedures, including microbial containment measures of a facility for "Wilding mice" using blood sampling for immunophenotyping as an example.
08:30
Immunophenotyping of Orthotopic Homograft (Syngeneic) of Murine Primary KPC Pancreatic Ductal Adenocarcinoma by Flow Cytometry
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Cited by 9 •
2018
The experimental procedure on the immunophenotyping of murine orthotopic PDAC homografts aims at profiling the tumor immuno-microenvironment. Tumors are orthotopically implanted via surgery. Tumors of 200–600 mm3 in size were harvested and dissociated to prepare single-cell suspensions, followed by multi-immune marker FACS analysis using different fluorescently-labeled antibodies.
FAQs
Marker selection determines which immune-cell distinctions immunophenotyping can resolve. Antibodies are chosen for their ability to bind characteristic proteins associated with particular cell populations or functional states. Examining several markers together provides a more detailed profile than relying on a single signal, helping distinguish lymphocytes, monocytes, and other leukocytes within a sample.
Surface and intracellular markers answer different biological questions. Surface proteins help classify cells according to features exposed on the cell exterior, whereas intracellular markers provide information from within the cell. Including both types can connect a cell’s identity with aspects of its function, which is useful when immune composition alone does not explain a response.
Fluorescent labeling converts antibody binding into signals that can be measured for individual cells. Flow cytometry then considers those fluorescence signals alongside physical properties, allowing cells with different marker-expression patterns to be distinguished rather than treating the sample as a single average. This individual-cell analysis supports precise comparison of immune populations.
A basic workflow begins by selecting markers relevant to the immune populations or response under study, applying antibodies carrying fluorescent labels, and analyzing the labeled cells with flow cytometry. The resulting measurements are interpreted by comparing marker expression and physical properties among individual cells. This links the experimental design directly to the biological question.
Immunophenotyping is especially informative when researchers need to follow changing leukocyte populations across infection, disease progression, or treatment. Immune profiles can show whether lymphocyte, monocyte, or other leukocyte representation changes under these conditions. In infection studies, such comparisons help investigate how host defenses respond to pathogens rather than only recording pathogen presence.
The method can support diagnosis, infection monitoring, and broader immune profiling, but its value depends on interpreting marker patterns in context. A measured shift may reflect altered cell composition, changes in marker expression, or both. Linking these measurements with disease stage, treatment, or host-response questions helps turn a cellular profile into biologically meaningful evidence.