Cells were cyto-spun onto glass slides at about 0.1e6 concentration per spot at 600RPM for 4 minutes and air-dried for 10 minutes at room temprature. Slides were then stained in May-Grünwald Stain (Sigma) for 5 minutes, followed by a 1.5 minute rinse in Phosphate buffer (sigma). Slides were placed for 20 minutes in Geimsa solution (Sigma) diluted 1:20 with DI water and rinsed for 2 minutes in DI water. After staining, cells were dried at room temperature, mounted in Cytoseal 60 (Thermo Scientific) and submitted for imaging on Zeiss AX10 microscope.
Geimsa solution
Manufactured by Merck Group
Geimsa solution is a staining reagent commonly used in microscopy and laboratory analysis. It is a mixture of methylene blue, eosin Y, and azure B in a buffered solution. The primary function of Geimsa solution is to stain cellular structures, such as nuclei and cytoplasm, to enhance their visibility and contrast under a microscope.
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4 protocols using geimsa solution
1
Cytospin Staining for Cell Analysis
2
Clonogenic Assay of BM-MSCs
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To evaluate the clonogenicity, the BM-MSCs were plated at a density of 350 cells/9.01 cm2 culture dish (TPP). After incubation for 9 days, the colonies formed were fixed by methanol (Sigma-Aldrich) and stained with Geimsa solution (Sigma-Aldrich) [33 ]. CFU numbers were enumerated by a light microscope and a cluster of at least 20 cells was defined as a CFU.
3
Cytospin Staining for Cell Analysis
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Cells were cyto-spun onto glass slides at about 0.1e6 concentration per spot at 600RPM for 4 minutes and air-dried for 10 minutes at room temprature. Slides were then stained in May-Grünwald Stain (Sigma) for 5 minutes, followed by a 1.5 minute rinse in Phosphate buffer (sigma). Slides were placed for 20 minutes in Geimsa solution (Sigma) diluted 1:20 with DI water and rinsed for 2 minutes in DI water. After staining, cells were dried at room temperature, mounted in Cytoseal 60 (Thermo Scientific) and submitted for imaging on Zeiss AX10 microscope.
4
Cell Migration Assay Protocol
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Serum starved cells (5x10 4 ) were seeded onto a semi-permeable membrane inserts (8μm pore size) with 10% FBS medium below. After 24h, inserts were washed with PBS followed by fixation with 3.7% Formaldehyde and permeabilization with 100% Methanol. Inserts were then washed and stained with Geimsa solution (Sigma, 48900). Non-migrated cells were removed using a cotton swab and images were taken using EVOS FL Cell Imaging System (4 fields for each cell type) and analyzed using Fiji software.
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