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Incucyte software

Manufactured by Sartorius
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The IncuCyte software is a live-cell imaging and analysis system that enables real-time monitoring and quantification of cellular processes. It provides objective, quantitative data on cell health, proliferation, and other cellular parameters in a non-invasive manner.

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Lab products found in correlation

Woundmaker, by Sartorius (10 mentions) Incucyte s3, by Sartorius (9 mentions) Incucyte live cell imager, by Sartorius (9 mentions) Incucyte, by Sartorius (8 mentions) Incucyte cytotox green reagent, by Sartorius (4 mentions) Countess automated cell counter, by Thermo Fisher Scientific (4 mentions) Incucyte s3 live cell analysis system, by Sartorius (3 mentions) Imagelock tissue culture microplate, by Sartorius (3 mentions) Incucyte cytotoxicity assay, by Sartorius (3 mentions) Annexin 5 pi staining, by BD (3 mentions) Vybrant dyecycle green stain, by Thermo Fisher Scientific (3 mentions) Incucyte live cell analysis system, by Sartorius (3 mentions) Incucyte s5 live cell imaging system, by Sartorius (2 mentions) Incucyte imagelock 96 well plate, by Sartorius (2 mentions) Prism 6, by GraphPad (2 mentions) Incucyte zoom system, by Sartorius (2 mentions) Incucyte zoom live cell analysis system, by Sartorius (2 mentions) Geltrex, by Thermo Fisher Scientific (2 mentions) Wst 1, by Agilent Technologies (2 mentions) Wst 1, by Promega (2 mentions)

85 protocols using incucyte software

1

Cell Cycle and Apoptosis Analysis

Cited 1 time
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Annexin V-PI staining (BD Biosciences, San Jose, CA, USA) was used to measure cell cycle progression and apoptosis induction as previously described [45 (link)]. Caspase 3/7 apoptosis activity was measured using Incucyte software (Essen Biosciences, Ann Arbor, MI, USA) as previously described [38 (link)]. Briefly, fluorescence of caspase 3/7 substrate was divided by the total number of cells measured using Vybrant® DyeCycle™ Green stain (Life Technologies, Grand Island, NY, USA) to obtain the apoptotic index. Data were analyzed using Incucyte software (Essen Biosciences).
Braggio D., Zewdu A., Londhe P., Yu P., Lopez G., Batte K., Koller D., Costas Casal de Faria F., Casadei L., Strohecker A.M., Lev D, & Pollock R.E. (2020). β-catenin S45F mutation results in apoptotic resistance. Oncogene, 39(34), 5589-5600.
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2

Quantifying Cell Proliferation and Colony Formation

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Cell proliferation assay was measured with WST-1 (promega) reagent
according to the manufacturer’s instruction (Clontech). Briefly,
cells were treated with WST-1 for 2hours at 37oC incubator prior to
absorbance reading at 440nm using the KC4 microplate reader (BioTek). Each
absorbance was normalized to the media control without any cells. For the
Incucyte cell confluence assay, C4–2B cells were infected with
pLKO.1V, shEZH2, shSUZ12 or shAR for 24 hours and harvested by
trypsinization. 5,000 cells were counted on a Countess automated cell
counter (Life Technologies, Carlsbad, CA) and plated on 24 tissue culture
plates in 3 replicates. Photomicrographs were taken every two hours using an
Incucyte live cell imager (Essen Biosciences, Ann Arbor, MI). Cell
confluence were measured using Incucyte software (Essen Biosciences, Ann
Arbor, MI) over 5 days in culture. Data were normalized to the pLKO.1
control cells and analyzed using Incucyte software (Essen Biosciences, Ann
Arbor, MI).
For colony formation assay, 1,000–2,000 cells were plated in
each well of a 6-well plate and treated with indicated concentration of
DMSO, Enz, EPZ or both for 10–14 days, cells were fixed by 4%
paraformaldehyde and stained by 0.05% crystal violet.
Kim J., Lee Y., Lu X., Song B., Fong K.W., Cao Q., Licht J.D., Zhao J.C, & Yu J. (2018). Polycomb- and Methylation-Independent Roles of EZH2 as a Transcription Activator. Cell reports, 25(10), 2808-2820.e4.
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3

Apoptosis Induction and Cell Cycle Analysis

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Cell cycle progression and apoptosis induction using Annexin V-PI staining (BD Biosciences) were measured as previously described [28 (link)]. Caspase 3/7 apoptosis activity was measured using Incucyte software (Essen Biosciences) as previously described (Braggio, Cancer, 2019). Briefly, apoptotic index was measured by dividing the fluorescence of caspase 3/7 substrate by total number of cells measured using Vybrant® DyeCycle™ Green stain (Life Technologies). Data were analyzed using Incucyte software (Essen Biosciences).
Braggio D.A., Costas C. de Faria F., Koller D., Jin F., Zewdu A., Lopez G., Batte K., Casadei L., Welliver M., Horrigan S.K., Han R., Larson J.L., Strohecker A.M, & Pollock R.E. (2022). Preclinical efficacy of the Wnt/β-catenin pathway inhibitor BC2059 for the treatment of desmoid tumors. PLoS ONE, 17(10), e0276047.
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4

Quantifying Cell Proliferation and Colony Formation

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Cell proliferation assay was measured with WST-1 (promega) reagent
according to the manufacturer’s instruction (Clontech). Briefly,
cells were treated with WST-1 for 2hours at 37oC incubator prior to
absorbance reading at 440nm using the KC4 microplate reader (BioTek). Each
absorbance was normalized to the media control without any cells. For the
Incucyte cell confluence assay, C4–2B cells were infected with
pLKO.1V, shEZH2, shSUZ12 or shAR for 24 hours and harvested by
trypsinization. 5,000 cells were counted on a Countess automated cell
counter (Life Technologies, Carlsbad, CA) and plated on 24 tissue culture
plates in 3 replicates. Photomicrographs were taken every two hours using an
Incucyte live cell imager (Essen Biosciences, Ann Arbor, MI). Cell
confluence were measured using Incucyte software (Essen Biosciences, Ann
Arbor, MI) over 5 days in culture. Data were normalized to the pLKO.1
control cells and analyzed using Incucyte software (Essen Biosciences, Ann
Arbor, MI).
For colony formation assay, 1,000–2,000 cells were plated in
each well of a 6-well plate and treated with indicated concentration of
DMSO, Enz, EPZ or both for 10–14 days, cells were fixed by 4%
paraformaldehyde and stained by 0.05% crystal violet.
Kim J., Lee Y., Lu X., Song B., Fong K.W., Cao Q., Licht J.D., Zhao J.C, & Yu J. (2018). Polycomb- and Methylation-Independent Roles of EZH2 as a Transcription Activator. Cell reports, 25(10), 2808-2820.e4.
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5

Measuring Cell Cycle and Apoptosis

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Cell cycle progression was measured as previously described 21 (link). Apoptosis was measured using Annexin V-PI staining (BD Biosciences) as previously described 21 (link). Caspase 3/7 apoptosis activity was measured using Incucyte software (Essen Biosciences) over 15 days in culture. Apoptotic index was measured by dividing the fluorescence of caspase 3/7 substrate by total number of cells measured using Vybrant® DyeCycle™ Green stain (Life Technologies). Data were analyzed using Incucyte software (Essen Biosciences).
Braggio D., Koller D., Jin F., Siva N., Zewdu A., Lopez G., Batte K., Casadei L., Welliver M., Strohecker A.M., Lev D, & Pollock R.E. (2019). AUTOPHAGY INHIBITION OVERCOMES SORAFENIB RESISTANCE IN S45F-MUTATED DESMOID TUMORS. Cancer, 125(15), 2693-2703.
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6

Cell Viability and Apoptosis Assay

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Cells were harvested by trypsinization, counted on a Countess automated cell counter (Life Technologies, Carlsbad, CA) and plated at 2,000 cells per well on 96 tissue culture plates in 3 replicates. Cells were treated with the indicated reagents after cells were attached to the plate at 24 hours post initial incubation. Photomicrographs were taken every two hour using an Incucyte live cell imager (Essen Biosciences, Ann Arbor, MI). Confluence and caspase3/7 apoptosis activity were measured using Incucyte software (Essen Biosciences, Ann Arbor, MI) over 3 days in culture. Apoptotic index was measured by dividing the number of caspase 3/7 positive cells as measured by fluorescence of caspase 3/7 substrate by total number of cells as measured using Vybrant Green DyeCyle (Life Technologies, Carlsbad, CA) at 3 days post indicated treatment. Data were analyzed using Incucyte software (Essen Biosciences, Ann Arbor, MI).
Lee Y., Wang Y., James M., Jeong J.H, & You M. (2015). Inhibition of IGF1R Signaling Abrogates Resistance to Afatinib (BIBW2992) in EGFR T790M Mutant Lung Cancer Cells. Molecular carcinogenesis, 55(5), 991-1001.
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7

Quantifying Cell Adhesion and Proliferation

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One hundred microliters per well of the POS conjugate were added to RPE cells in triplicate. Samples were preincubated for 1 to 2 h at 37 °C with 5% of CO2. Wells were washed with 100 µL of EDTA, followed by three washes with PBS. After the last wash, 100 µL/well of DMEM FluoroBrite containing 2 mM HEPES was added. Plates were then placed in the plate holder of an IncuCyte® located in a 37 °C, 5% CO2 incubator for IncuCyte scanning or scanned once using the ImageXpress® Micro Confocal (IMX) system. The plates were scanned with the phase contrast and orange channel of the IncuCyte instrument for 24 h. After scanning, the data were analyzed using the IncuCyte software (Sartorius, Gottingen, Germany). The total orange object’s integrated intensity (Y-axis) was generated from each sample as a function of time (X-axis). The plates were scanned around the peak time with the red and green channels of the IMX.
Liang H., Wu Q., Guo X.V., Chan L., Mao T., Stella C., Guilbaud A, & Camperi J. (2023). Comprehensive Analysis of Photoreceptor Outer Segments: Flow Cytometry Characterization and Stress-Driven Impact on Retinal Pigment Epithelium Phagocytosis. International Journal of Molecular Sciences, 24(16), 12889.
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8

Detecting SIE in A. aegypti cells

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To test whether SIE can be detected in A. aegypti cells, Aag2 cells were infected with SINV mCherry at three different MOI (0.1, 1, and 10), and after 24 h of incubation, a secondary infection with SINV ZsGreen was carried out. For each of the infections, the inoculum was left on the cell monolayer for 1 h and subsequentially exchanged for fresh L-15 media. Plates were imaged using the IncuCyte live cell imager (Sartorius) after 24 h of incubation following the secondary infection and images analyzed using the associated IncuCyte software (Sartorius). The experiment was carried out in eight technical replicates per treatment.
Reitmayer C.M., Levitt E., Basu S., Atkinson B., Fragkoudis R., Merits A., Lumley S., Larner W., Diaz A.V., Rooney S., Thomas C.J., von Wyschetzki K., Rausalu K, & Alphey L. (2023). Mimicking superinfection exclusion disrupts alphavirus infection and transmission in the yellow fever mosquito Aedes aegypti. Proceedings of the National Academy of Sciences of the United States of America, 120(37), e2303080120.
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9

Organoid-based High-throughput Drug Screening

Cited 3 times
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We have used a human tumor sample to establish pancreatic and ovarian organoids. And for lung organoids, we have used tumor samples from an autochthonous mouse model (KrasG12D+/, p53R172H+/—, AdCre).80 (link) Tumor samples were chopped and digested enzymatically using digestion media containing collagenase II, Dispase, and 1% FBS. After digestion, the cells were embedded in Matrigel and incubated in the incubator for 15 to 20 min before adding organoids-specific media.22 (link),81 (link),82 (link)Three independent replicates of human ovarian and pancreatic organoids and murine lung organoids were transfected with siRNA (for PAF1/PD2), followed by treatment with cisplatin or gemcitabine alone or in combination with RAD52 inhibitor D-I03. Real-time images of the organoids were captured every 3 h for 48–72 h using the IncuCyte-S5 live-cell imaging system (Sartorius). The kinetic data (organoid average growth and darkness) were analyzed and graphically represented using IncuCyte software (Sartorius). RNA was collected from organoids at the experimental endpoint to perform further analysis.
Rauth S., Ganguly K., Atri P., Parte S., Nimmakayala R.K., Varadharaj V., Nallasamy P., Vengoji R., Ogunleye A.O., Lakshmanan I., Chirravuri R., Bessho M., Cox J.L., Foster J.M., Talmon G.A., Bessho T., Kishor Ganti A., Batra S.K, & Ponnusamy M.P. (2023). Elevated PAF1-RAD52 axis confers chemoresistance to human cancers. Cell reports, 42(2), 112043.
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10

Real-time Apoptosis and Cell Death Assay

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Cells were treated with respective drugs as indicated, followed by staining the cells with IncuCyte Annexin V Orange dye (Sartorius, 4759) or Annexin V Green dye (Sartorius, 4624) for apoptosis or with IncuCyte cytotox green reagent for counting dead cells (Sartorius, 4633). Fluorescent objects were quantified in real-time to identify apoptosis or cell death via IncuCyte integrated analysis software (Sartorius). Phase-contrast images taken on the same vessels were used to quantify cell proliferation or growth using the IncuCyte cell-by-cell analysis software module (Sartorius). Real-time images of the cells were captured every 3 h for 48–72 h and were analyzed and graphically presented using IncuCyte software (Sartorius).
Rauth S., Ganguly K., Atri P., Parte S., Nimmakayala R.K., Varadharaj V., Nallasamy P., Vengoji R., Ogunleye A.O., Lakshmanan I., Chirravuri R., Bessho M., Cox J.L., Foster J.M., Talmon G.A., Bessho T., Kishor Ganti A., Batra S.K, & Ponnusamy M.P. (2023). Elevated PAF1-RAD52 axis confers chemoresistance to human cancers. Cell reports, 42(2), 112043.
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