Incucyte zoom

Manufactured by Sartorius

Sourced in United States, United Kingdom, Germany, Japan

The IncuCyte ZOOM is a live-cell analysis system that enables real-time monitoring and quantification of cellular processes. It provides automated, non-invasive, and time-lapse imaging capabilities to observe and measure cellular responses within a controlled environment.

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Lab products found in correlation

Woundmaker, by Sartorius (30 mentions) Imagelock plate, by Sartorius (20 mentions) Sytox green, by Thermo Fisher Scientific (12 mentions) Matrigel, by Corning (12 mentions) Incucyte s3, by Sartorius (11 mentions) Imagelock 96 well plate, by Sartorius (11 mentions) Incucyte zoom software, by Sartorius (10 mentions) 96 well plate, by Corning (10 mentions) Incucyte woundmaker, by Sartorius (9 mentions) Matrigel, by BD (8 mentions) Woundmaker tool, by Sartorius (7 mentions) Incucyte, by Sartorius (6 mentions) Incucyte caspase 3 7 green apoptosis assay reagent, by Sartorius (6 mentions) 96 well woundmaker, by Sartorius (6 mentions) Incucyte zoom system, by Sartorius (6 mentions) 96 well plate, by Greiner (6 mentions) Incucyte kinetic caspase 3 7 apoptosis assay reagent, by Sartorius (5 mentions) Woundmaker device, by Sartorius (5 mentions) Imagelock, by Sartorius (5 mentions) Sytox green nucleic acid stain, by Thermo Fisher Scientific (5 mentions)

661 protocols using incucyte zoom

Real-Time Monitoring of Nanoparticle-Treated Cells

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For live cell imaging, cells treated with Lipo-PLEKHA7-PEG-cRGD nanoparticles were compared with untreated cells. KG-1a cells were seeded into 96-well plates (at a density of ∼4 × 10⁴ cells/well), incubated for 4 h, and then treated with Lipo-PLEKHA7-PEG-cRGD. After washing twice with PBS, the cells were suspended in fresh media and were monitored in the IncuCyte ZOOM (Essen BioScience), acquiring images at 1 h scan intervals for 48 h.
For cell growth assessment, KG-1a and ML-2 cells were seeded into 96-well plates (at a density of ∼4 × 10⁴ cells/well), incubated for 4 h, and then treated with Lipo-PLEKHA7-PEG-cRGD. After washing twice with PBS, the cells were stained with IncuCyte NucLight Rapid Red Reagent for nuclear labeling (Essen Bioscience) and were monitored in the IncuCyte ZOOM (Essen BioScience), acquiring images at 1 h scan intervals for 48 h.
For cell growth quantification, transfected cells and untreated cells were seeded in 96-well plates (5000 cells/well) using IncuCyte ZOOM (Essen Bioscience) for every 1 h of imaging. The confluence was analyzed using the IncuCyte ZOOM 2016A software.

Mohammed S.A., Kimura Y., Toku Y, & Ju Y. (2022). Bioengineered PLEKHA7 nanodelivery regularly induces behavior alteration and growth retardation of acute myeloid leukemia. Biomaterials and Biosystems, 6, 100045.

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Automated Wound Healing Assay

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Scratch assay were performed, and cell migration was monitored by the automated Incucyte ZOOM (Sartorius, Gottingen, Germany). 67NR cells infected with the FN mutant plasmids were seeded in an Essen Imagelock 96-well plate at the density of 1×60⁴ cells/well and were maintained over night to reach 100% confluency. A wound was made in the cell monolayer using a 96-well wound-maker tool with PTFE pin tips (Sartorius) according to the manufacturer’s instructions. Afterward, the plate was inserted into the Incucyte ZOOM and incubated in 5% CO2 at 37°C for 53 h. Images were captured at 1-h intervals, and data were analyzed using the Incucyte ZOOM software (Sartorius). Relative wound density (RWD), which measures the relative cell density in the wounded and non-wounded areas at each time point, was used to assess the rate of cell migration.

Ernst W.G., Maruyama S, & Wallis S. (1997). Buoyancy-driven, rapid exhumation of ultrahigh-pressure metamorphosed continental crust. Proceedings of the National Academy of Sciences of the United States of America, 94(18), 9532-9537.

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Automated Wound Healing Assay

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Melamed S., Zaffryar-Eilot S., Nadjar-Boger E., Aviram R., Zhao H., Yaseen-Badarne W., Kalev-Altman R., Sela-Donenfeld D., Lewinson O., Astrof S., Hasson P, & Wolfenson H. (2023). Initiation of fibronectin fibrillogenesis is an enzyme-dependent process. Cell reports, 42(5), 112473.

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Cell Growth and Apoptosis Assays

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For growth assays, cells were plated in 96-well plates at 5,000 or 7,500 cells/well and transfected with 2% (v/v) of non-perturbing nuclear-restricted green fluorescent label (IncuCyte NucLight Green BacMam 3.0, Essen Bioscience). For apotosis assays, cells were plated in 96-well plates at 10,000 cells/well and transfected with 1% (v/v) apoptosis marker reagent, which is cleaved by activated caspase 3/7, releasing a green fluorescent label (IncuCyte Caspase-3/7 Apoptosis Assay Reagent). After 2 h, cells were incubated in an Incucyte Zoom (Essen Bioscience), acquiring phase and green fluorescent images were obtained at 10X magnification every 2 h. Incucyte Zoom software was used to analyze and graph the results.

Magani F., Peacock S.O., Rice M.A., Martinez M.J., Greene A.M., Magani P.S., Lyles R., Weitz J.R, & Burnstein K.L. (2017). Targeting AR Variant-coactivator Interactions to Exploit Prostate Cancer Vulnerabilities. Molecular cancer research : MCR, 15(11), 1469-1480.

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Breast Cancer Cell Migration Assay

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First, a 50-μl quantity of Matrigel (100 μg/ml, BD Bioscience) was added onto an Image Lock 96-well plate (Essen BioScience) overnight at 37°C in 5% CO₂. Then, 6 × 10⁴ DCIS^alone2cy, DCIS^cnt2cy, and DCIS^CAF2cy were seeded onto the Matrigel-precoated plates in 100 μl of DMEM/F12 with 5% FBS for 4 h at 37°C in 5% CO₂ (Figs 2B and 5C). The scratch wound was also generated by the Wound Maker tool (Essen BioScience), before washing with PBS once and coating with 50 μl of Matrigel (8 mg/ml) on a prechilled cool box. To solidify the Matrigel, the plate was subsequently incubated at 37°C in 5% CO₂ for 30 min followed by addition of 100 μl of 5% FBS in DMEM/F12 at 37°C in 5% CO₂. The plate was scanned with an IncuCyte Zoom (Essen BioScience) according to the manufacturer’s instructions at 3-h intervals for 5 d.
Plates were incubated for 1 h and then treated with DMSO, saracatinib (1 μM), or PP1 (10 μM) (Fig 6H). The plates were scanned with an IncuCyte Zoom (Essen BioScience) according to the manufacturer’s instructions at 2-h intervals for 4 d.

Matsumura Y., Ito Y., Mezawa Y., Sulidan K., Daigo Y., Hiraga T., Mogushi K., Wali N., Suzuki H., Itoh T., Miyagi Y., Yokose T., Shimizu S., Takano A., Terao Y., Saeki H., Ozawa M., Abe M., Takeda S., Okumura K., Habu S., Hino O., Takeda K., Hamada M, & Orimo A. (2019). Stromal fibroblasts induce metastatic tumor cell clusters via epithelial–mesenchymal plasticity. Life Science Alliance, 2(4), e201900425.

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Quantitative Cell Migration Analysis

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U2OS cells were reversely transfected in IncuCyte ImageLock 96-well plates (Essen Bioscience) and grown into confluent monolayers before wounding with the IncuCyte WoundMaker (Essen Bioscience). Imaging was performed by using the IncuCyte ZOOM with a 10× objective (Essen Bioscience), and the relative wound density calculated by the IncuCyte ZOOM software analysis program. For individual cell tracking and analysis of cell speed, the Fiji/ImageJ manual tracking plugin and Ibidi Chemotaxis software was used.

Bjørnestad S.A., Guadagno N.A., Kjos I, & Progida C. (2022). Rab33b-exocyst interaction mediates localized secretion for focal adhesion turnover and cell migration. iScience, 25(5), 104250.

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Caspase-3/7 Activation Imaging Assay

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Cells were cultured in 96-well plates (5000 cells/well) and 0.1% IncuCyte Caspase-3/7 Green Reagent (Essen Bioscience) was added to the medium with PG or Gly to detect caspase 3/7-activated cells. Cells were assessed in triplicate in each condition. Immunofluorescent images were taken at 3 h intervals up to 24 h at 10 × magnification using IncuCyte Zoom (Essen Bioscience). Caspase 3/7-positive cells were counted using the IncuCyte Zoom software (Essen Bioscience).

Komura M., Sato T., Yoshikawa H., Nitta N.A., Suzuki Y., Koike K., Kodama Y., Seyama K, & Takahashi K. (2022). Propylene glycol, a component of electronic cigarette liquid, damages epithelial cells in human small airways. Respiratory Research, 23, 216.

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Quantifying Cell Growth and Apoptosis

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To monitor cell growth, the live cell imaging instrument IncuCyte ZOOM (Essen Bioscience) was used. Cells were plated in a 96-well Micro Greiner clear plate (Sigma-Aldrich) and imaged every 4 h with default software settings and a 10× objective. The IncuCyte software was used to quantify confluence from two non-overlapping bright-field images. To identify apoptotic cells, the IncuCyte ZOOM instrument was used in combination with the Cell Player 96-well kinetic caspase-3/7 reagent (Essen Bioscience). The software was used to calculate mean green fluorescence for two non-overlapping images of each well. Green fluorescence was normalized to phase-contrast confluence to determine apoptosis. For crystal violet assays, the cells were cultured in six-well dishes and fixed using 4% formaldehyde for 10 min. Plates were washed using running tap water and stained using 1 ml 0.1% crystal violet for 30 min exactly.

Benedict B., van Bueren M.A., van Gemert F.P., Lieftink C., Guerrero Llobet S., van Vugt M.A., Beijersbergen R.L, & te Riele H. (2020). The RECQL helicase prevents replication fork collapse during replication stress. Life Science Alliance, 3(10), e202000668.

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Caspase 3/7 Activation in Lung Myofibroblasts

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Kinetic estimation of caspase 3/7 activity was performed using the real-time imaging system IncuCyte ZOOM (Essen BioScience, Ann Arbor, MI, United States). Activation of caspase-3/7 in cells undergoing apoptotic death cleaves the caspase-3/7 substrate to produce nuclear green-fluorescence (Caspase-3/7 Green Apoptosis Assay Reagent [Essen Bioscience]). Primary lung-resident myofibroblasts or fibrocytes were prepared from normal or fibrotic lung tissue and cultured in a 12-well plate to 50–60% confluency. After growing overnight in low serum-containing MEM media, they had adapted to low-serum conditions. They were then treated with media containing either Caspase 3/7 Green Apoptosis Assay Reagent at a final concentration of 5 μM/mL or Caspase 3/7 Green Apoptosis Assay Reagent and anti-Fas antibody (BD Biosciences) at a final concentration of 250 ng/mL. Time-lapse fluorescence imaging was performed using the IncuCyte ZOOM system (Essen BioScience); 9 images per well at 20× magnification were collected every 2 h for 24–48 h. The average number of green objects produced by the apoptotic cells were measured using IncuCyte ZOOM software 2015A.

Kasam R.K., Reddy G.B., Jegga A.G, & Madala S.K. (2019). Dysregulation of Mesenchymal Cell Survival Pathways in Severe Fibrotic Lung Disease: The Effect of Nintedanib Therapy. Frontiers in Pharmacology, 10, 532.

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Proliferation, Motility, and Adhesion Assays

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For proliferation assays, cells were plated at 2,000 cells per well of a 96-well dish. Four pictures of different regions in each well were taken every four hours for 48 hours using an IncuCyte ZOOM (Essen BioScience). Percent confluence was determined using the IncuCyte ZOOM software (Essen BioScience). For motility assays, wells of a 96-well dish were coated with 200 mg/mL matrigel (Corning) and allowed to gel for 2 hours at 37°C. Liquid was then aspirated and replaced with 2 mg/mL bovine serum albumin and blocked for 1 hour at 37°C. Wells were then washed once with PBS and cells were plated to produce a uniform monolayer 24 hours later. A WoundMaker (Essen BioScience) was used to make scratches across each well. Scratches were cleaned by washing wells two times with PBS. Pictures were taken at the center of the scratch every hour for 30 hours, and the IncucyteZOOM software was used to measure percent wound closure over time. For adhesion assays cells were plated at 5,000 cells/well in a 96-well dish. 6 hours post-plating the plates were washed with PBS and the cells were fixed in 10% neutral buffered formalin before being stained with 10% crystal violet. Images of each well were taken for analysis of positive pixels using Image J.

Black S., Nelson A.C., Gurule N., Futscher B.W, & Lyons T.R. (2016). Semaphorin 7a exerts pleiotropic effects to promote breast tumor progression. Oncogene, 35(39), 5170-5178.

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