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

## TL;DR

This paper presents a protocol for staining semi-adherent multiple myeloma cells with minimal morphological distortion using immunofluorescence.

## Contribution

A new method for immunofluorescence staining of semi-adherent cells with reduced centrifugation steps.

## Key findings

- The protocol preserves fine cellular details in semi-adherent RPMI 8226 cells.
- Reducing centrifugation steps minimizes morphological distortions during staining.
- Fixing and permeabilizing cells in solution improves structural preservation.

## Abstract

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.

For complete details on the use and execution of this protocol, please refer to Osei-Amponsa et al.1

•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

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.

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.