Cell viability and VCD were measured using Vi-cell BLU Cell Viability Analyzer (Beckman Coulter). The measurement was done using either normal mode with 200 ± 20 μL of sample or a fast mode with 170 μL of sample.
Vi cell blu cell viability analyzer
Manufactured by Beckman Coulter
The Vi-CELL BLU Cell Viability Analyzer is a compact, automated device that measures cell viability and cell count. It utilizes trypan blue dye exclusion and image-based analysis to provide accurate and reliable results.
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9 protocols using vi cell blu cell viability analyzer
1
Cell Viability Measurement Using Vi-cell BLU
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Chlorite Cytotoxicity Assay in HeLa Cells
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HeLa cells were trypsinized, counted, and prepared at 105 cells/mL in normal growth media. One molar sodium chlorite stock solution was prepared fresh at the time of the assay in UltraPure ddH2O, and diluted to 2× working concentrations in cell growth media. Cells were seeded in 24-well plates, with triplicate wells for each condition. Five hundred microliters of each 2× chlorite/media preparation was first added to the plate. Five hundred microliters of cell suspension (5 × 104 cells) was then added to each well. The plate was gently mixed, and cells were grown for 3 d in a 37 °C/5% CO2 incubator. After 3 d, cells were washed briefly with 500 μL of PBS, trypsinized with 250 μL TrypLE Express, and resuspended with 750 μL of normal growth media to 1 mL total volume. In wells containing a majority of visibly dead, floating cells, cells were resuspended by vigorously pipetting up and down rather than by trypsinization. Two hundred microliters of each cell suspension was then quantified using a Vi-Cell BLU Cell Viability Analyzer (Beckman Coulter).
3
AREG Neutralization Impacts Cell Viability
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Cells were plated in their respective maintenance media conditions at a density of 50,000 cells/well in 12-well plates. 24 hours following seeding, cells were treated with 1, 3, or 5 μg of AREG neutralizing antibody (R&D Systems, MAB262-SP) or an equivalent volume of PBS. This experiment was performed with three replicates for each cell line and each treatment condition. Cells were counted and viability was measured 72 hours following treatment using a Vi-CELL BLU cell viability analyzer (Beckman Coulter).
4
Cell proliferation assay for MCF-10A knockouts
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Exponentially growing MCF-10A cells of each knockout genotype were plated at 2 × 103 cells per well in six-well plates. On indicated days, cells were counted using a Beckman Coulter Vi-Cell BLU Cell Viability Analyzer. All cell lines were counted in triplicate.
5
AREG Neutralization Impacts Cell Viability
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Cells were plated in their respective maintenance media conditions at a density of 50,000 cells/well in 12-well plates. 24 h following seeding, cells were treated with 1, 3, or 5 µg of AREG neutralizing antibody (R&D Systems, MAB262-SP) or an equivalent volume of PBS. This experiment was performed with three replicates for each cell line and each treatment condition. Cells were counted and viability was measured 72 h following treatment using a Vi-CELL BLU cell viability analyzer (Beckman Coulter).
6
Isolation and Analysis of Murine Splenocytes
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Recombinant MHV68 expressing H2bYFP fusion protein (rMHV68-H2bYFP) was used for the infection (107 (link)). Mice (8 to 12 weeks old) were infected by intraperitoneal injection with 1,000 PFU in 0.5 ml under isoflurane anesthesia. Mouse spleens were homogenized, treated to remove red blood cells, and passed through a 100-µm-pore-size nylon filter. For ex vivo experiments, flow sorting, and RNA-sequencing, splenocytes were subject to negative selection to enrich for B cells (Pan B cell isolation kit; STEMCELL, Vancouver, BC, Canada). Cells were counted using Vi-CELL BLU Cell Viability Analyzer (Beckman Coulter, Pasadena, CA). Blood was collected posthumously by cardiac puncture with gauge 27 needle. Serum was collected after a centrifugation at 20,000 x g for 20 mins at room temperature.
7
Cell Viability Assay Protocol
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Cell lines were maintained and treated as previously described (Cell culture with drug). 100,000 cells/well were plated in a 6-well plate, on Day 0, and cells began treatment 24 or 72 hours later (in accordance with Cell culture with drug). Three wells of cells were trypsinized and collected (per cell line and treatment) every 2 days. Cells were then counted via trypan blue exclusion using the Beckman Coulter Vi-CELL BLU Cell Viability Analyzer (Beckman Coulter Life Sciences).
8
Cell Viability Analysis of LAA + Olaparib
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Cell density data was acquired from cells treated with LAA + Olaparib for 72hr. Cells were sampled on a Vi-CELL BLU Cell Viability Analyzer (Beckman). 100 images acquired per sample, diameter 6–30μm, cell sharpness 7, minimum circularity, medium decluster degree, 3 aspiration and mixing cycles, 50% viable spot brightness, and 5% viable spot area.
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Cell Viability Assay with Trypan Blue
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