May-Grunwald Giemsa Staining: A Method to Stain Bone Marrow Cells

Published: April 30, 2023

Abstract

Source: Mopin, A. et al. A Detailed Protocol for Characterizing the Murine C1498 Cell Line and its Associated Leukemia Mouse Model. J. Vis. Exp. (2016).

This video describes the technique of staining bone marrow cells using May- Grünwald Giemsa Staining. In the protocol, we will stain the murine myeloid leukemia cells, C149 cells, to analyze the morphology and differential counting.

Protocol

1. In vitro Characterization of the C1498 Cell Line In vitro culture of C1498 cells Prepare complete RPMI (Roswell Park Memorial Institute) 1640 medium by adding 50 ml of fetal bovine serum (FBS), 5 ml of penicillin (100 U/ml)-streptomycin (100 µg/ml), 500 µl of 50 mM β-mercaptoethanol, 5 ml of N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid (HEPES), 5 ml of non-essential amino acids and 5 mL of sodium pyruvate to 500 ml of RPMI medium. <l…

Declarações

The authors have nothing to disclose.

Materials

C1498 cell line   ATCC TIB 49
2-Mercaptoethanol, Gibco  ThermoFisher Scientific 21985
Fetal Bovine Serum (FBS)  ThermoFisher Scientific 10270
HEPES, Gibco (1 M)   ThermoFisher Scientific 15630
Penicillin-Streptomycin, Gibco   ThermoFisher Scientific 15140
RPMI 1640 Medium (Gibco, GlutaMAX Supplement)  ThermoFisher Scientific 61870
Trypan blue solution (0.4 %)    ThermoFisher Scientific 15250-061 concentration 1/2
Chamber & filter card (EZ Cytofunnel Shandon)   Thermo Scientific A78710003
May-Grünwald solution   RAL Diagnostics 320070
Giemsa R solution    RAL Diagnostics 320310 concentration 1/20
pH 6.8 buffer solution  RAL Diagnostics 330368
Slides (Starfrost – ground edges 90)   Knittel Glass VS1137# 077FKB
Microscope Cover Glasses, 24 x 24mm   Knittel Glass VD1 2424 Y100
Shandon Cytospin 3 Cytocentrifuge  ThermoFisher Scientific

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May-Grunwald Giemsa Staining: A Method to Stain Bone Marrow Cells. J. Vis. Exp. (Pending Publication), e20264, doi: (2023).

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