Protocol for cytoskeleton staining of the semi-adherent multiple myeloma cell line RPMI 8226 by immunofluorescence

Highlights

• Protocol for immunofluorescence microscopy of semi-adherent cells
• Steps for preparing multiple myeloma RPMI 8226 cells for immunostaining
• Approach for minimal distortion of cell morphology by reducing centrifugation

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Summary

Preservation of fine cellular details of semi-adherent or suspension cells for imaging by immunofluorescence is challenging. This protocol enables staining of floating cells with minimal morphological distortions, as we demonstrate with the semi-adherent multiple myeloma cell line RPMI 8226. We describe steps to better preserve structural details by fixing, permeabilizing, and staining cells in solution, while minimizing the number of centrifugation steps and centrifugation g-force.

Subject areas

Cell Biology; Cell culture; High-Throughput Screening; Immunology; Microbiology; Antibody

Abdos Product Used

Author

Vasty Osei-Amponsa, Valentin Magidson, Kylie J. Walters

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