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Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
Evaluating B-cells: from bone marrow precursors to antibody-producing cells
M Manuela Rosado1, Marco Scarsella, Simona Cascioli
1Research Center Ospedale Pediatrico Bambino Gesù (IRCCS), Rome, Italy.
Methods in Molecular Biology (Clifton, N.J.)
|August 15, 2013
Summary
Flow cytometry (FACS) and ELISA techniques identify murine B-cell subsets. These methods quantify antibody-secreting cells (ASC), crucial for antibody production in bone marrow and peripheral tissues.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Lymphocyte identification relies on surface marker expression.
- Flow cytometry (FACS) is key for B-cell subset identification, function, and activation analysis.
- B cells differentiate into antibody-secreting cells (ASC).
Purpose of the Study:
- To combine FACS and ELISA techniques for identifying murine B-cell subsets.
- To quantify antibody-secreting cells (ASC) in bone marrow and peripheral tissues.
Main Methods:
- Flow cytometry analysis (FACS) for B-cell subset identification.
- Enzyme-linked immunosorbent assay (ELISA) techniques, including Enzyme-linked immunosorbent spot (ELISPOT) assay.
- Analysis of murine bone marrow and peripheral B-cell populations.
Main Results:
- Successful identification of murine bone marrow and peripheral B-cell subsets using FACS.
- Quantification of antibody-secreting cells (ASC) number and specificity via ELISPOT assay.
- Demonstration of FACS and ELISA synergy in B-cell characterization.
Conclusions:
- Combined FACS and ELISA provide a robust method for murine B-cell subset characterization.
- ELISPOT assay effectively measures ASC, vital for antibody production.
- This approach enhances understanding of B-cell differentiation and antibody responses.
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