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Cells on microspheres: a new technique for flow cytometric analysis of adherent cells
Abstract:
Technical problems have previously prevented the application of fluorescence activated cell sorting to the study of adherent cell populations. We have developed a procedure for attachment of human monocyte-macrophages to 14-20 micrometer microspheres. These adherent cells on microspheres retained phagocytic capacity, could be stained for cell surface antigens using indirect immunofluorescence, and could be maintained in long-term culture. They could be examined and sorted on a fluorescence activated cell sorter while still in the adherent state. This technique facilitates the flow cytometric study of adherent cells and permits the isolation of subpopulations of these cells.
Insights
Researchers developed a method to study adherent cells using fluorescence activated cell sorting (FACS). This technique allows for the analysis and isolation of specific cell subpopulations, overcoming previous technical limitations.
Area of Science:
- Cell Biology
- Immunology
- Biotechnology
Background:
- Adherent cell populations have been difficult to study using fluorescence activated cell sorting (FACS) due to technical limitations.
- Previous methods were not suitable for analyzing cells that naturally adhere to surfaces.
Purpose of the Study:
- To develop a novel procedure for applying FACS to the study of adherent cell populations.
- To enable the analysis and isolation of subpopulations within adherent cell types.
Main Methods:
- Attachment of human monocyte-macrophages to 14-20 micrometer microspheres.
- Staining of cells on microspheres for cell surface antigens using indirect immunofluorescence.
- Examination and sorting of adherent cells on a fluorescence activated cell sorter.
Main Results:
- Adherent cells on microspheres maintained their phagocytic capacity.
- Cells could be successfully stained for surface antigens and maintained in long-term culture.
- Successful examination and sorting of adherent cells using FACS was achieved.
Conclusions:
- The developed procedure overcomes previous technical barriers in studying adherent cells with FACS.
- This technique facilitates flow cytometric analysis of adherent cells.
- It enables the isolation of specific subpopulations of adherent cells for further study.