A total of 1.5 × 103 PC cells were seeded in six-well plates and treated with different concentrations (0, 20, 40, and 80 μM) of ST. After culturing for 10 days, cell colonies were immobilized with 4% paraformaldehyde and stained with 0.5% crystal violet (Boster, Wuhan, Boster). Thereafter, a stereogram and micrograph of the colony plates were obtained using a camera and optical microscope (magnification 40×), respectively.
Crystal violet
Manufactured by Boster Bio
Sourced in China
Crystal violet is a synthetic dye used as a staining agent in various laboratory applications. It is primarily used to stain biological samples, such as cells or tissues, to enhance their visibility and contrast under a microscope. Crystal violet is a key component in the Gram staining procedure, which is used to differentiate between Gram-positive and Gram-negative bacteria.
Automatically generated - may contain errors
9 protocols using crystal violet
1
Impact of ST on PC Cell Colony
2
Cell Viability Assay with T4O
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A total of 5 × 103 PC cells were placed in six-well plates. Different concentrations (0, 0.5, 1, and 2 μM) of T4O were used to treat the cells. Each treatment group consisted of three replicates. After culturing for 10 days, the cell colonies were immobilized with 4% paraformaldehyde and stained with 0.5% crystal violet (Boster, Wuhan, China). The experiments were performed in triplicate.
3
Paracrine Effects of HSCs on Tumor Invasion
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In order to assess the paracrine effects of HSCs on tumor invasion and migration, LX2 cells with or without NRP-1 knockdown were serum starved and CM were collected. The Transwell chambers (pore size, 8.0 µm; EMD Millipore, Billerica, MA, USA) without (for the migration assay) or with Matrigel (for the invasion assay; BD Biosciences, Franklin Lakes, NJ, USA) coatings were inserted into a 24-well culture plate.
For the migration assay, the HepG2 cells (100 µl, 5×104) suspended in DMEM supplemented with 1% FBS were placed in the upper chamber and 0.5 ml CM collected from LX2-NRP-1 shRNA, LX2-NT shRNA and LX2-control was added into each lower chamber as a chemoattractant. The Transwell chambers were then incubated for 24 h.
For the invasion assay, 8-µm pore chamber inserts were coated with Matrigel. HepG2 cells in the log phase of growth were cultured in 6-well plates (100 µl; 5×105/ml) in medium supplemented with 1% FBS for 24 h. The remaining steps were the same as for the migration assay. The Transwell chambers were incubated for 48 h.
The migrated and invaded cells on the underside of the filter were fixed in 37% methanol and stained with crystal violet (Boster Biological Technology, Pleasanton, CA, USA). Cell migration and invasion was determined by counting the stained cells in 10 randomly selected fields using a light microscope (magnification, ×100).
For the migration assay, the HepG2 cells (100 µl, 5×104) suspended in DMEM supplemented with 1% FBS were placed in the upper chamber and 0.5 ml CM collected from LX2-NRP-1 shRNA, LX2-NT shRNA and LX2-control was added into each lower chamber as a chemoattractant. The Transwell chambers were then incubated for 24 h.
For the invasion assay, 8-µm pore chamber inserts were coated with Matrigel. HepG2 cells in the log phase of growth were cultured in 6-well plates (100 µl; 5×105/ml) in medium supplemented with 1% FBS for 24 h. The remaining steps were the same as for the migration assay. The Transwell chambers were incubated for 48 h.
The migrated and invaded cells on the underside of the filter were fixed in 37% methanol and stained with crystal violet (Boster Biological Technology, Pleasanton, CA, USA). Cell migration and invasion was determined by counting the stained cells in 10 randomly selected fields using a light microscope (magnification, ×100).
4
Cell Colony Formation Assay
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NB cells were inoculated in six-well plates (1000 cells/well) and treated with different concentrations (0, 5, and 10 μM) of CYN. After culturing for 14 days, cell colonies were immobilized with 4% paraformaldehyde for 30 min and stained with 0.5% crystal violet (Boster, Wuhan, Boster) for 15 min.
5
Cell Migration and Invasion Assay
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Cells were suspended in medium without FBS and a total of 1 × 105cells were then added to the upper chambers, which were pre-coated with (for invasion assay) or without (for migration assay) Matrigel (BD Biosciences). Medium supplemented with 20% FBS was added to the lower chamber. Cells were cultured at 37 °C for another 48 h, after which, the migrated/invaded cells were fixed with 4% paraformaldehyde and stained using 0.5% crystal violet (Boster Biological Technology, Wuhan, China) at room temperature for 30 min. After washing with PBS, the chambers were air-dried and observed under an inverted light microscope (Olympus, Tokyo, Japan).
6
Transwell Assay for Invasion and Migration
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Transwell assay was employed to detect cell invasion and migration according to the manufacturer's protocol (BD Biosciences, Bedford, MA, USA). After 24 h of transfection, cells were resuspended in FBS-free DMEM, 100 μL of DMEM medium containing cells (5×104 for SCC-9,1×105 for CAL-27) were added to the upper chamber, and supplemented with 600 μL DMEM medium containing 10% FBS in the lower chamber. After 24 h of incubation, cells were fixed with methanol for 30 min and were stained with 0.1% crystal violet (Boster, Wuhan, China). Five fields were randomly selected for cell counting under microscope.
7
Transwell Migration and Invasion Assay
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For transwell migration assay, total 4´10 5 cells were suspended using 200μl DMEM medium without FBS and seeded into the upper transwell chambers (Becton, Dickinson and Company, USA). 600μl DMEM medium contained 10% FBS was placed in the lower transwell chambers. After 28h, migratory cells were xed with paraformaldehyde and stained using 0.5% crystal violet. The transwell invasion experiment required matrigel(Becton, Dickinson and Company, USA) to be coated in the upper chamber before cell seeding. After xation using 4% paraformaldehyde, staining with 1% crystal violet (Boster, Wuhan, China) and removing non-invasion cells using cotton swab. Finally, chambers were was photoed and counted.
8
Crystal Violet Assay for Cell Viability
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Cell vitality after drug treatment and siRNA transfection was evaluated by crystal violet assay. 48 h after treatment, PBS (phosphate-buffered saline) was used to wash cells (, which were then xed by methanol for 15 min, followed by staining with one percent crystal violet (BOSTER, China) for 20 min. Following washing with PBS, cells were dried on lter paper, and pictures were taken by a scanner. Then one millilitre 1% SDS was added in each well, and crystals were completely dissolved by shaking the plate on a low-speed shaker for 15 min. The A 570 was determined using a microplate reader.
9
Transwell Migration and Invasion Assay
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For transwell migration assay, total 4´10 5 cells were suspended using 200μl DMEM medium without FBS and seeded into the upper transwell chambers (Becton, Dickinson and Company, USA). 600μl DMEM medium contained 10% FBS was placed in the lower transwell chambers. After 28h, migratory cells were xed with paraformaldehyde and stained using 0.5% crystal violet. The transwell invasion experiment required matrigel(Becton, Dickinson and Company, USA) to be coated in the upper chamber before cell seeding. After xation using 4% paraformaldehyde, staining with 1% crystal violet (Boster, Wuhan, China) and removing non-invasion cells using cotton swab. Finally, chambers were was photoed and counted.
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