VSV-SARS-CoV-2-S Δ21 viral infection rates were imaged and quantified using the Incucyte® S3 Life Cell Analysis System. VeroE6, H522, and H522-ACE2 cells were labeled with Incucyte® NucLight Red (Sartorius #4625) to generate stable expression of the red nuclear marker. Basal airway epithelial cells (AEC) were labeled with Incucyte® NucLight Rapid Red (Sartorius #4717) at the time of infection. VSV-SARS-CoV-2-S Δ21 was then added to cells and immediately placed in the Incucyte®. Phase and fluorescent images were taken every hour to track viral infection. Percentage of GFP positive cells was calculated by dividing green object count by red object count for each well.
Incucyte nuclight rapid red
Manufactured by Sartorius
The Incucyte® NucLight Rapid Red is a live-cell analysis instrument designed for real-time monitoring and quantification of cell growth, proliferation, and other cellular processes in in vitro cell cultures. The device utilizes fluorescent-based detection and proprietary image analysis algorithms to provide quantitative data on the cell population dynamics.
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Lab products found in correlation
Incucyte nuclight red, by Sartorius (2 mentions)
Incucyte s3, by Sartorius (2 mentions)
Incucyte analysis software, by Sartorius (1 mentions)
Incucyte s3 live cell imaging system, by Sartorius (1 mentions)
Incucyte caspase 3 7 reagent, by Sartorius (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
5 protocols using incucyte nuclight rapid red
1
Quantifying VSV-SARS-CoV-2-S Δ21 Infection
2
High-throughput hepatocyte imaging analysis
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Primary hepatocytes were plated in 12-well plates at a density of 2∗105cells/well. Medium was changed every other day, and treatments or controls were added as described in text to appropriate wells. Treated cells were imaged by IncuCyte Live-Cell Analysis System for 24 hours with imaging at ×20 magnification. All 12 treatments were imaged in 4 unique fields per treatment every 30 minutes using a nuclear stain (IncuCyte NucLight Rapid Red; Sartorius, Essen Bioscience, Ann Arbor, MI), caspase-GFP–labeled constructs, cell death marker Annexin V (Annexin V NIR; Sartorius), phase contrast, and green (400 ms exposure) channels in IncuCyteS3 (Sartorius) platform, housed at 37°C with 5% CO2.
3
Real-Time Viral Infection Imaging
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VSV-SARS-CoV-2-SΔ21 viral infection rates were imaged and quantified using the Incucyte® S3 Life Cell Analysis System. VeroE6, H522, and H522-ACE2 cells were labeled with Incucyte® NucLight Red (Sartorius #4625) to generate stable expression of the red nuclear marker. Basal airway epithelial cells (AEC) were labeled with Incucyte® NucLight Rapid Red (Sartorius #4717) at the time of infection. VSV-SARS-CoV-2-SΔ21 was then added to cells and immediately placed in the Incucyte®. Phase and fluorescent images were taken every hour to track viral infection. Percentage of GFP positive cells was calculated by dividing green object count by red object count for each well. Normalized GFP Intensity was calculated by dividing green intensity by percent confluence.
4
Time-course Apoptosis Assays
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Time-course apoptosis assays following either 5AZA or decitabine treatment were performed using an IncuCyte® S3 (Essen Bioscience). Cells were seeded at a density of 20,000 cells/well. The reagents used to label the cells were IncuCyte® Nuclight Rapid Red and Cell Health Reagent Caspase-3/7 (both used at a 1:2,000 final dilution in the wells; Essen Bioscience). Cells were incubated in a humidified incubator at 37°C with 5% CO2. Each condition was performed in triplicate.
5
Kinetic Apoptosis Imaging Assay
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For kinetic image-based assays, tumor cells were seeded in a volume of 180 μL with Incucyte® NucLight Rapid Red (Essen Biosciences) and Incucyte Caspase 3/7 reagent (Essen Biosciences). One to three images/day/well were acquired using an Incucyte S3 live cell imaging system (Essen Biosciences). Image acquisition began 1 day after seeding and coincided with the addition of cortisol/relacorilant (day 1; 0 h). Paclitaxel (Sigma) was added on day 2 (24 h). Image analysis was conducted using the Incucyte Analysis Software (Essen Biosciences) to determine the number of cells positive for each fluorescence marker. The apoptotic index was calculated by dividing the number of caspase 3/7 positive cells by the total number of NucLight positive cells.
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