The invasion ability of breast cancer cells in vitro was evaluated by Matrigel coated Transwell inserts (BD Biosciences, San Diego, CA, USA). 1 × 105 cells in 200 μL FBS‐free medium were added in the upper chamber of Transwell and 10% FBS‐containing medium was added in the lower chamber. After 16 h, the cells were fixed by 4% paraformaldehyde and stained by Giemsa stain (Solarbio, Beijing, China). The cells were then observed under a microscope and the number of migrating cells was counted in five predetermined fields.
Giemsa stain
Manufactured by Solarbio
Sourced in China
Giemsa stain is a laboratory reagent used for the staining of cells and tissues. It is a polychromatic stain that selectively colors different cellular components, enabling the visualization and identification of various cell types and structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Solarbio (4 mentions)
Paraformaldehyde (pfa), by Sangon (3 mentions)
Trypsin edta, by Solarbio (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Soft agar, by BD (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
β catenin, by Santa Cruz Biotechnology (2 mentions)
N cadherin, by Santa Cruz Biotechnology (2 mentions)
P gsk3β, by Santa Cruz Biotechnology (2 mentions)
Cck 8 reagent, by BestBio (2 mentions)
E cadherin, by Santa Cruz Biotechnology (2 mentions)
Matrigel coated transwell insert, by BD (1 mentions)
Colchicine, by Merck Group (1 mentions)
Colchicine, by Thermo Fisher Scientific (1 mentions)
Digital camera, by Sony (1 mentions)
Matrigel, by Corning (1 mentions)
Axio observer a1 microscope, by Zeiss (1 mentions)
24 well transwell plate, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Transwell chamber, by Corning (1 mentions)
32 protocols using giemsa stain
1
Evaluating Breast Cancer Cell Invasion
2
Colony Formation Assay for HepG2, SK-Hep-1 and BEL 7402 Cells
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For colony formation assays, approximately 200 infected HepG2, SK-Hep-1 and BEL 7402 cells were seeded into six-well plates and cultured for 14 days at 37°C. After removal of the medium and washing with PBS, the cells were fixed in 4% paraformaldehyde for 20 minutes and then stained with Giemsa stain (Solarbio, Beijing, China). Only positive colonies (> 50 cells/colony) in the plates were counted and compared.
3
Evaluating Breast Cancer Cell Invasion
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The invasion ability of breast cancer cells in vitro was evaluated by Matrigel coated Transwell and Transwell inserts (BD Biosciences, San Diego, CA, USA). 1 × 105 cells in 200 μl FBS-free medium were added in upper chamber of transwell and 10% FBS contained medium was added in lower chamber. After 16 hours, the cells were fixed by 4% paraformaldehyde and stained by Giemsa stain (Solarbio). Then the cells were observed under a microscope and the number of migrating cells was counted in five predetermined fields.
4
Radiosensitivity Evaluation of SCLC
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A colony formation assay was utilized to evaluate the radiosensitivity of SCLC cells. Briefly, cells were plated into 6-well plates and treated with different treatments. Chemical reagents were removed at 72 h after IR, and the cells were cultured for 8–12 days to develop colonies and stained using Giemsa stain (Solarbio). Colonies with >50 cells were used to calculate the survival fraction.
5
Karyotypic Analysis of ImGIST Cells
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The ImGIST cells were treated with 0.25 μg/mL colchicine (Sigma-Aldrich, St. Louis, MO, USA) for 1–2 h at 37 °C, followed by cell collection. Cells were incubated in a hypotonic solution for 30 min at 37 °C and then fixed in methanol/glacial acetic acid (v/v = 3:1) for 10 min. The cells were digested with trypsin, fixed, and then stained with Giemsa stain (Solarbio). Mitotic cells with good dispersion and moderate staining were selected for karyotypic observation and microscopic analysis.
6
Giemsa Staining of Transduced Cells
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Gene transduced A375 and B16 cells were seeded into 6-well plates (2 × 103 cells/ well) and cultured for 14 days. Then the cells were first washed twice with PBS solution and fixed with 4% paraformaldehyde (Solarbio) for 15 min. The cells were stained using Giemsa stain (Solarbio).
7
Transwell Invasion Assay Protocol
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We used a transwell plate with an 8μm diameter polycarbonate film to evaluate the invasion ability of cells. Then, 70μl Matrigel matrix was applied on the upper layer of the membrane 2 hours in advance. The cells were resuspended with serum-free medium and planted on the Matrigel matrix (5×104 cells per well), and the complete medium was used in the lower compartment. After incubation for 24-72 h, the cells were fixed with methanol, stained with 0.5% Giemsa stain (Solarbio, China) and then photographed and counted under the microscope.
8
Evaluating Cell Proliferation with PSPD3R
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The cells were plated at a low density and incubated with increasing PSPD3R concentrations. At the end of the growth period, the cells were fixed with methyl alcohol, stained with Giemsa stain (Solarbio, Beijing, China), and photographed to estimate their proliferation relative to that of untreated cells.54
9
Karyotype Analysis Protocol
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For karyotype analysis, colchicine (Gibco-Life Technology, USA) was added at 0.06 μg/mL for 6 h before the cell suspension was harvested. Then attached cells were dissociated, and cell suspensions were added into 0.075 M hypotonic KCl for 30 min. The cells were fixed with 1:3 acetic acid:methanol at 4°C overnight. Finally, the prepared cells were stained with Giemsa stain (Beijing Solarbio Science & Technology, China) for 10 min and examined under a microscope (Leica Microsystems GmbH, Germany).
10
Cell Proliferation Assay Protocol
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Cells were seeded at a density of 500 cells/well in six-well cell culture plates at 37°C for 14 days. After taking photographs and washing with phosphate-buffered saline (PBS; Solarbio, Baijing, China), the cells were fixed for 30 min by adding 1 mL of 4% paraformaldehyde (Solarbio) to each well. Subsequently, the fixed cells were stained for 15 min using 500 μL of Giemsa stain (Solarbio). Then, photographs were taken with a digital camera (Sony, Tokyo, Japan) and clones were counted.
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