Spontaneous migration assay was performed as described previously [39 (link)]. Briefly, 250 cells were seeded in a complete medium into each well of the dedicated 96-well plate (IncuCyte ImageLock, Sartorius, Goettingen, Germany), and the plates were incubated in an IncuCyte Live Cell Analysis Imaging System (Sartorius, Goettingen, Germany). Series of plate images were collected from 0 to 72 h every 2 h. Collected images were analyzed with a Manual Tracking plugin (ImageJ, F. Cordelieres, Institute Curie, Paris, France).
Incucyte imagelock
Manufactured by Sartorius
Sourced in Germany, United Kingdom
The Incucyte ImageLock is a specialized piece of laboratory equipment designed for live-cell imaging applications. It provides a controlled and stable environment for cells, enabling researchers to capture high-quality images and monitor cellular behavior over extended periods of time.
Automatically generated - may contain errors
Lab products found in correlation
Woundmaker, by Sartorius (2 mentions)
Incucyte zoom software, by Sartorius (2 mentions)
Incucyte live cell analysis imaging system, by Sartorius (1 mentions)
Incucyte live cell analysis system, by Sartorius (1 mentions)
Matrigel, by Corning (1 mentions)
Incucyte scratch wound cell migration software module, by Sartorius (1 mentions)
Incucyte woundmaker, by Sartorius (1 mentions)
Mitomycin c, by Merck Group (1 mentions)
96 well woundmaker, by Sartorius (1 mentions)
7 protocols using incucyte imagelock
1
Spontaneous cell migration assay
2
Cell Migration Tracking Protocol
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To measure the distance and speed of cell migration, CRC cells from co-culture or mono-culture were seeded on 96-well plates (IncuCyte ImageLock, Sartorius) covered with Matrigel (1 mg/ml) (Corning) and incubated for 1 h at 37 °C. Then, phase-contrast time-lapse images were captured for 24 h with time intervals of 2 h using a 10 × objective in an IncuCyte® Live-Cell Analysis System (Sartorius). Pictures were analyzed with ImageJ software and the Manual Tracking plugin [13 (link)]. The distance covered by every cell was measured as the total distance based on the cumulative track lengths. Two independent experiments were carried out, and in each one, at least 25 cells were analyzed.
3
Matrigel-based Scratch Wound Assay
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Cancer cells derived from co-culture or mono-culture were seeded into Matrigel-coated (0.5 mg/ml) ImageLock 96-well plates (IncuCyte ImageLock, Sartorius) and incubated for 24 h to reach 100% confluence. Then, standardized scratches were made in all wells simultaneously using Wound Maker™ (Essen Bioscience). Subsequently, cells were covered with an additional layer of Matrigel, on top of which a 1:1 combination of culture and conditioned medium (with FBS collected as described in the “Co-culture conditions ” section) was added into each well. Phase-contrast time-lapse images were taken every 2 h for 24 h using an IncuCyte® LiveCell Analysis System using a 10 × objective. Representative results were analyzed using the IncuCyte® Scratch Wound Cell Migration Software Module (Sartorius). The relative wound density represents the increase in the area covered by the cells over time. The experiments were performed three times, and each independent experiment consisted of four repetitions.
4
Cell Confluence Measurement by Incucyte
5
Wound Healing Assay with Incucyte
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Approximately 22,500 eACs/cm2 (P3) were seeded per 96-well plate well (Incucyte Image Lock, Essen Bioscience, Ltd., Royston, UK) in the presence of HG-DMEM medium containing 10% FCS and PSA. Twenty-four hours post-seeding, treatments were added. Thirty hours post-seeding (time = 0 h), the scratch was performed with the Incucyte WoundMaker (Essen Bioscience). Two washing steps with PBS were performed to remove detached cells. Finally, the treatments were added. The relative wound density was assessed, corresponding to the ratio of the area colonized by the cells (time = t) to the total area of the initial scratched region (time = 0 h).
6
Wound Healing Dynamics in HaCaT Cells
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HaCaT cells (1.5 × 104 cells/well) were seeded onto a collagen I-coated 96-well IncuCyte™ ImageLock™ tissue culture plate (Essen BioScience, catalog no. 4379) and incubated in a standard CO2 incubator for 48 h to form a cell monolayer, before being treated with 2 µg/mL Mitomycin C (Sigma-Aldrich, catalog no. M0503) for 2 h. Wounds were made with the 96-well WoundMaker™ (Essen BioScience, 4493). In order to remove any detached cells, wounded cell layers were washed twice with culture medium before treatment with 100 μl of medium containing the appropriate glucose concentrations. Images of the wounds were automatically acquired within the CO2 incubator using the IncuCyte™ ZOOM software package (Essen BioScience, catalog no. 2016A). Typical kinetic updates were taken at 3 h intervals for the duration of the experiment. Finally, cell confluence analysis was performed using the IncuCyte™ ZOOM software.
7
NLRP3 Inflammasome-Induced Migration in HCASMC
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HCASMC (1.0 × 104 cells/well) were seeded in 8 replicates onto 96-well IncuCyte ImageLock tissue culture plates (Essen BioScience, #4379) and incubated for 72 h to form a cell monolayer, before being treated with NLRP3-YFP inflammasome particles (3:1 particles/ cell) for 4 h. PDGF (10 ng/ml) and Actinomycin (5 µg/ml) were incubated for 24 h as positive and negative controls. Wounds were made with the 96-well WoundMaker (Essen BioScience, #4493). The plate was washed twice to remove any detached cells. Images of the wounds were automatically acquired within the CO2 incubator using the IncuCyte ZOOM software package (Essen BioScience, #2016A). Typical kinetic updates were taken at 0 and after 24 h. Wound confluency [%] was calculated using the IncuCyte ZOOM software. Cell count that migrated into the scratch were counted manually.
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