50,000 – 100,000 cells were harvested, washed once at 300 × g for 5 min, resuspended in 200 μLof FACS Buffer, and spun onto poly-L-lysine coated microscope slides with a Shandon 4 (Thermo Scientific) cytocentrifuge at 300 rpm for 4 min. When visibly dry slides were transferred into May-Grünwald solution (Sigma-Aldrich) for 5 min, rinsed 4 times for 30 s in water, and transferred to Giemsa solution (Sigma-Aldrich) for 15 min. Slides were washed as described above, dry mounted with coverslips, and examined. All images shown were taken using a Metafer slide scanning platform and software (Metasystems) at 63X magnification.
May gr nwald solution
Manufactured by Merck Group
Sourced in Germany, United States
May-Grünwald solution is a staining reagent used in microscopy and hematology laboratories. It is a mixture of eosin and methylene blue dyes that stains cellular components, allowing for the visualization and differentiation of different cell types in blood smears and other biological samples. The core function of May-Grünwald solution is to provide a standardized staining protocol for the identification and examination of cells under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Giemsa solution, by Merck Group (21 mentions)
Shandon 4, by Thermo Fisher Scientific (3 mentions)
Metafer slide scanning platform and software, by MetaSystems (3 mentions)
Evos xl core imaging system, by Thermo Fisher Scientific (3 mentions)
Giemsa, by Merck Group (2 mentions)
Shandon cytospin 3, by Thermo Fisher Scientific (2 mentions)
Cytospin system, by Thermo Fisher Scientific (1 mentions)
Ds ri1, by Nikon (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
Dxm1200f digital camera, by Nikon (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
Shandon cytospin 4, by Thermo Fisher Scientific (1 mentions)
Dm2000 inverted microscope, by Leica (1 mentions)
Dxm1200f, by Nikon (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Giemsa stain solution, by Merck Group (1 mentions)
Ckx53, by Olympus (1 mentions)
Neutral buffered formalin solution, by Merck Group (1 mentions)
May gr nwald giemsa, by Merck Group (1 mentions)
Zymosan, by Merck Group (1 mentions)
32 protocols using may gr nwald solution
1
Cell Preparation for Microscopy Analysis
2
Cell Staining Using Cytospin and Giemsa
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Cells were attached to glass slides using the Cytospin system (Thermo Scientific). Samples were air dried and then sequentially stained with May-Grünwald solution (Sigma-Aldrich) and Giemsa solution (Sigma-Aldrich). Images were captured with a Nikon Eclipse 80i equipped with a CCD camera DS-Ri1 (Nikon), and analyzed using Photoshop CS6 (Adobe).
3
Cell Preparation for Microscopy Analysis
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50,000 – 100,000 cells were harvested, washed once at 300 × g for 5 min, resuspended in 200 μLof FACS Buffer, and spun onto poly-L-lysine coated microscope slides with a Shandon 4 (Thermo Scientific) cytocentrifuge at 300 rpm for 4 min. When visibly dry slides were transferred into May-Grünwald solution (Sigma-Aldrich) for 5 min, rinsed 4 times for 30 s in water, and transferred to Giemsa solution (Sigma-Aldrich) for 15 min. Slides were washed as described above, dry mounted with coverslips, and examined. All images shown were taken using a Metafer slide scanning platform and software (Metasystems) at 63X magnification.
4
Cell Morphology Evaluation via Microscopy
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Cell morphology was evaluated using light optical microscopy using both the crystal violet and May–Grünwald–Giemsa stainings. The cells were grown in glass coverslips. After treatment, crystal violet staining was performed as previously described for the DNA content evaluation. For the May–Grünwald–Giemsa staining, the cells were stained with a May–Grünwald solution (Sigma-Aldrich, St. Louis, MO, USA) for 3 min and then a Giemsa solution (Sigma-Aldrich, St. Louis, MO, USA) for 15 min. After rinsing with distilled water, the coverslips were allowed to dry at room temperature. Representative images were acquired using a Nikon Eclipse 80i microscope equipped with a Nikon Digital Camera DXM 1200F at 100× and 500× magnification. Analysis was performed in duplicate and repeated in at least three independent experiments.
5
Tissue Fixation and Staining Protocol
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Tissues were fixed in 10% neutral-buffered formalin for 24 h and then stored in 70% ethanol. Soft tissues were mounted in paraffin, sectioned, and stained by hematoxylin and eosin. Peripheral blood smears were fixed with methanol for 1 min. May Grünwald–Giemsa stain was performed using the May Grünwald solution (Sigma-Aldrich 32856) and the modified Giemsa stain (Sigma-Aldrich GS1L) according to the manufacturer's recommendations.
6
Cytospin Staining and Imaging Protocol
7
Microscopic Assessment of Cell Death
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Optical microscopy was used to assess the morphological features associated with apoptosis and necrosis. A quantity of 1 × 106 cells was collected and seeded in glass slides. Then, smears were stained for 3 min with May–Grünwald solution (Sigma-Aldrich, St. Louis, MO, USA) and for 15 min with Giemsa solution (Sigma-Aldrich). After rinsing with distilled water, cell morphology was analyzed using light microscopy with a Nikon Eclipse 80i microscope equipped with a Nikon digital camera DXm 1200F (Nikon, Tokyo, Japan).
8
Bovine Muscle Cell Differentiation Assay
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Calf and mature cattle muscle cells were seeded at 3 x 10⁴ cells/mL in a 12-well plate to assess myogenic cell differentiation. When the cells reached 80–90% confluence, the medium was changed to promote differentiation of the cells. The differentiation media (DM) contained DMEM-F12 with 2% horse serum (Gibco, Gaithersburg, USA), and 1% A.A.
When the cells began to differentiate, their morphology was examined at 0 h. Cell differentiation was assessed after 24, 48, 72, and 96 h. The cells were stained with May-Grünwald solution (Sigma-Aldrich, MO, USA) and Giemsa stain solution (Sigma-Aldrich). After removing the medium, cells were washed twice with DPBS. Then, 100% methanol was added for cell fixation and the cells were allowed to stand for 10 min. Then, methanol was removed and the May-Grünwald solution was added for 5 min. Deionized water (D.W) was added for dilution. After removing the solution, Giemsa staining solution was added (1:20) for 20–30 min. Finally, the solution was removed and the stained cells were observed under a microscope (CKX53, OLYMPUS, Tokyo, Japan).
When the cells began to differentiate, their morphology was examined at 0 h. Cell differentiation was assessed after 24, 48, 72, and 96 h. The cells were stained with May-Grünwald solution (Sigma-Aldrich, MO, USA) and Giemsa stain solution (Sigma-Aldrich). After removing the medium, cells were washed twice with DPBS. Then, 100% methanol was added for cell fixation and the cells were allowed to stand for 10 min. Then, methanol was removed and the May-Grünwald solution was added for 5 min. Deionized water (D.W) was added for dilution. After removing the solution, Giemsa staining solution was added (1:20) for 20–30 min. Finally, the solution was removed and the stained cells were observed under a microscope (CKX53, OLYMPUS, Tokyo, Japan).
9
May-Grünwald Giemsa Staining Protocol
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For May-GrÜ nwald Giemsa stain, cytocentrifuged cells were stained with May-GrÜ nwald solution (Cat# MG500, Sigma-Aldrich, St. Louis, MO) for 5 minutes and in Giemsa (Cat# GS500, Sigma-Aldrich, St. Louis, MO) for 20 minutes. For whole bone marrow sections, femurs were fixed in 10% (vol/vol) neutral buffered formalin solution (NBF, approximately 4% formaldehyde) (Sigma-Aldrich, St. Louis, MO). The embedded sections were stained with hematoxylin and eosin (HistoServ, Germantown, MD).
10
Tissue Fixation and Blood Smear Staining
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Tissues were fixed in 10% formalin, embedded in paraffin, sectioned and stained with hematoxylin-eosin. Blood smears were fixed with 100% methanol and stained with May-Grünwald solution (Sigma, St. Louis, MO) and Giemsa (Sigma) solution.
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