A Convenient Technique to Fix Suspension Cells on a Coverslip for Microscopy

Current Protocols in Cell Biology
Keiko MiharaHisato Saitoh

Abstract

Human myeloid HL-60 cells are usually cultured in suspension in medium containing 5% to 10% fetal bovine serum (FBS) and thus are often difficult to adhere to a coverslip. In this unit, we describe how removal of FBS from the culture medium facilitates adhesion of HL-60 cells to coverslips. Importantly, HL-60 cells that adhere to the coverslips immersed in FBS-free medium can be immobilized in situ by conventional chemical fixatives and thus permeabilized for probing cellular structures using specific dyes and/or reagents, followed by microscopic observation. All-trans-retinoic-acid-exposed differentiated HL-60 cells, which have properties similar to neutrophils, can also adhere efficiently to coverslips in FBS-free medium. Because the procedure is not complex and special equipment is not required, the simplicity and cost effectiveness of this FBS-free cell adhesion protocol may be beneficial to researchers who are interested in assessing the structure and function of suspension cells using microscopy.

References

Aug 20, 2004·BMC Cell Biology·Ada L Olins, Donald E Olins
Mar 17, 2009·Nature Cell Biology·J Bradley ZucheroR Dyche Mullins
Mar 28, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Ingrid HepperBarbara Walzog
Nov 5, 2013·Methods in Enzymology·Cheryl M Koh
Nov 5, 2013·Methods in Enzymology·Cheryl M Koh
Aug 3, 2014·Analytical Biochemistry·Tomofumi NakayamaHisato Saitoh

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