Neurite outgrowth was quantified as described previously [14 (link)]. Briefly, cells were plated and treated in 96-well plates. After 4 days, cells were placed into the ZOOM IncuCyte Imaging System (Essen BioScience), and images were taken under 20X magnification. The relative neurite length was then calculated using the neurite definition determined for each specific cell line using the NeuroTrack system.
Incucyte zoom imaging system
Manufactured by Sartorius
Sourced in United States, United Kingdom
The IncuCyte ZOOM imaging system is a real-time, automated live-cell analysis platform. It enables continuous, non-invasive monitoring and analysis of cell behavior and biological processes within a cell culture environment.
Automatically generated - may contain errors
Lab products found in correlation
Woundmaker, by Sartorius (4 mentions)
Neurotrack software module, by Sartorius (3 mentions)
S7020, by Thermo Fisher Scientific (3 mentions)
Microplate reader, by Molecular Devices (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Krtap5 5 shrna resistant vector, by GenScript (2 mentions)
Imagelock 96 well plate, by Sartorius (2 mentions)
Imagelock plate, by Sartorius (2 mentions)
Incucyte s3, by Sartorius (2 mentions)
Prism 8, by GraphPad (2 mentions)
Cultureone supplement, by Thermo Fisher Scientific (1 mentions)
Woundmaker tool, by Sartorius (1 mentions)
Incucyte v2018b, by Sartorius (1 mentions)
Incucyte caspase 3 7 green dye for apoptosis, by Sartorius (1 mentions)
Incucyte woundmaker, by Sartorius (1 mentions)
Z vad fmk, by Peptide Institute (1 mentions)
Pd98059, by ChemScene (1 mentions)
Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions)
α smooth muscle actin α sma, by Agilent Technologies (1 mentions)
Cool snap, by Teledyne (1 mentions)
45 protocols using incucyte zoom imaging system
1
Automated Neurite Outgrowth Quantification
2
Time-lapse Neurite Outgrowth Imaging
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Cells were plated and treated in 96-well plates. For detecting neurite outgrowth, cells were placed into ZOOM IncuCyte Imaging System (Essen BioScience) and cell images were taken under 20X microscopic magnification. For detecting neurite outgrowth in a time-dependent manner, cell images were taken every 6 h. The neurite lengths associated with each treatment were calculated using the neurite definition defined for each specific cell line using the NeuroTrack system (Essen BioScience).
3
Quantifying Neurite Outgrowth after Gene Knockdown
4
Assessing Neurite Outgrowth in NPCs
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To assess neurite outgrowth, NPCs were plated at a density of 15,000 cells/cm2 in NeuroDiff media with modifications (substituting N2 with CultureOne supplement (Thermo Fisher)). The plate was imaged using an IncuCyte Zoom Imaging system (Essen Bioscience, Ann Arbor, MI) every 2 h for 7 days. Live-capture measurements were performed in 9 image field per well, n = 4 per genotype per treatment condition. For metformin treatment, metformin was added at day 1 post plating, and media with/without metformin was changed every 2–3 days. Cells were imaged under phase contrast, and analysis was performed using IncuCyte’s NeuroTrack module. The neurite-calling algorithm automatically traces the neurites from each cell body and masks the neurites for the entire image field, with no pre-selection of evaluated neurons. The growth rate of neurites in each well was obtained by measuring the surface area covered by neurites and expressed as mm/mm2.
5
In Vitro Wound Healing Assay with Dermal Fibroblasts
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Adult human dermal fibroblasts (HDF, Gibco) were plated at confluence (10,000 cells/well) in a 96‐well Image‐Lock plate (Essen Bioscience) and serum‐deprived overnight in medium containing 1% FBS. The following day, a standardized cell‐free lane (scratch) was created in each well using a WoundMaker tool (Essen Bioscience). The cell‐free lanes were washed with basal medium to remove cell fragments and remaining serum and subsequently treated with one of the following: basal medium without serum (negative control), complete medium with 10% FBS (positive control), and dHUC extracts at 20 mg/mL in basal medium (n = 10 donors). Soluble dHUC extracts were prepared as previously described.
The plates were transferred to the IncuCyte ZOOM imaging system (Software Version 2016B; Essen Bioscience) and imaged every 6 h for a total of 72 h. Percent wound confluence was determined using an image processing algorithm to differentiate HDF cells and cellular debris from background to avoid the influence of user bias during image analysis.
The plates were transferred to the IncuCyte ZOOM imaging system (Software Version 2016B; Essen Bioscience) and imaged every 6 h for a total of 72 h. Percent wound confluence was determined using an image processing algorithm to differentiate HDF cells and cellular debris from background to avoid the influence of user bias during image analysis.
6
Real-Time Cell Viability Monitoring
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To track cell viability in real time, BMDMs were stimulated in presence of the SYTOX Green nuclear stain that penetrates compromised membranes (1 μM; S7020; Life Technologies). Cell death was monitored over 22 h using the IncuCyte Zoom imaging system (Essen Biosciences) and data was collected using IncuCyte v2018B.
7
Quantifying Chlamydia-Induced Apoptosis
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IncuCyte HeLa Nuclight Red cells (Essen Biosciences) were seeded into 96-well plates at a density of 7.5 × 103 cells per well 24 h before infection with WT C. trachomatis L2 or InaC KO C. trachomatis L2 at an MOI of 1. The infection was synchronized by centrifugation at 1,000 × g for 1 h at room temperature. At 6 hpi, the IncuCyte caspase 3/7 green dye for apoptosis (Essen Biosciences) was added to each well at a final concentration of 5 μM. Wells were imaged every 2 h until 72 hpi using an IncuCyte Zoom imaging system (Essen Biosciences). The percentage of caspase 3/7-positive cells was calculated by dividing the number of caspase 3/7 puncta by the number of red nuclei.
8
Tracking Cell Death via NHE Stimulation
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To track viability, BMDMs or various cell types were stimulated with 0.5 μM NHE (high) or 100 nM NHE (low), LPS + ATP, or with 100 nM NHEWT or NHEmut., in presence of the SYTOX Green nuclear stain that penetrates compromised membranes (1 μM; S7020; Life Technologies). Cell death was monitored over 3 h using the IncuCyte Zoom imaging system (Essen Biosciences) or fluorescence was assessed using a Zeiss Axio Observer Fluorescent/Brightfield microscope.
9
Cytoprotective Effects of L-Cysteine Against MGO-Induced Toxicity
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MES13 and HEK 293 cells were seeded at 1.5 × 104 and 5.0 × 104 cells/well in 96-well plates and incubated for 24 h. The cells were pretreated with several concentrations (0.1, 0.5, and 1.0 mM) of l -cysteine and AG (1.0 mM) for 1 h, followed by treatment with MGO (500 μM) for 24 h. After incubation, the MTT (0.1 mg/mL) solution was added to each well and the cells were incubation for 2 h. Next, the medium was carefully removed and dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) was added. The absorbance was measured at 570 nm using a microplate reader (Molecular Devices, San Jose, CA, USA). Cell morphological changes in mesangial cells were investigated using the IncuCyte Zoom imaging system (Essen Bioscience, Ann Arbor, MI, USA).
10
Measuring Apoptosis in BxPC-3 Cells
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Control and S2/3-KO BxPC-3 cells were infected with the Nuclight lentivirus reagents (Sartorius #4625) in order to stained nucleus in red. 5 × 104 cells were seeded in 48-wells plate. The next day, cells were serum starved (1% FBS medium), treated or not with TGF-β and cleaved-Caspase 3/7 Green dye (Sartorius #4440) was added in the culture medium to follow apoptosis. Images were acquired and analyzed for 72 h with an Incucyte ZOOM Imaging System (Essen Bioscience).
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