Please use one of the following formats to cite this article in your essay, paper or report. Professor andrzej lange, lower silesian center for. Cell proliferation in cocultures using incucyte system and. We recommend optimizing incucyte nuclight rapid red reagent for each cell line tested by diluting the reagent in growth media final dilutions of 1. The incucyte nuclight rapid red reagent for nuclear labeling is a cell permeable dna stain that specifically stains nuclei in cells and is ideally suited to the mixandread, realtime quantification using the incucyte livecell analysis system. The incucyte nuclight rapid nir reagent for nuclear labeling is a cell. Incucyte is set to a 2hour scan pattern for everybody, you cannot change this. Cancers free fulltext the rnabinding ubiquitin ligase. Alternatively, cells can also labeled with our nuclight rapid red or nir nuclear binding fluorescent dyes. Mar 24, 2017 imelapse movie of coculture cell proliferation. What is the optimal seeding density and concentration of incucyte nuclight rapid red for u2 os cell proliferation assay. Incucyte livecell analysis handbook by sartorius issuu. At 100m and 10m ionomycin shows rapid onset of cytotoxicity within 612 hours of treatment. Incucyte cytotox reagents are highly sensitive cyanine nucleic acid dyes ideally suited to a simple mixandread, realtime quantification of cell death.
Incucyte cytolight rapid red reagent for live cell. Nuclight rapid red reagent is nonperturbing and enables highefficiency labeling. Downloading and connecting to softwareincucytezoom software can be downloaded from garvan software server. Incucyte nuclight lentivirus reagents provide homogenous expression of a nuclearrestricted gfp green fluorescent protein or mkate2 red fluorescent protein in your choice of primary. Incucyte, automated longterm livecell imaging system, offers a powerful platform for stem cell research. Incucyte annexin v green reagent for apoptosis incucyte. Essen bioscience launches the incucyte s3 livecell analysis. One vial of lyophilized solid sufficient for 100200 livecell apoptosis tests in 96well format. The incucyte rapid red reagent for cell labeling is a cell permeable dna stain that specifically stains nuclei in live cells and is ideally suited to the mixandread, realtime quantification of cell counting. Incucyte nuclight lentivirus reagents provide homogenous expression of a nuclearrestricted gfp green fluorescent protein or mkate2 red fluorescent protein in your choice of primary, immortalized, dividing, or nondividing cells without altering. Incucyte zoom technical details university of bristol. Four images per well were collected every 2 hr for 3 days using a 10. Incucyte cytolight rapid dyes for livecell labelling enable simple, fast and highly efficient labelling of living cells. Addition of the incucyte nuclight rapid red reagent to normal healthy cells is nonperturbing to cell growth.
I plan to assay proliferation dynamics of u2 os cells using the. Specially formulated, these reagents are ideally suited for real time measurements of apoptosis in living cultures. Mar 19, 2014 incucyte is set to a 2hour scan pattern for everybody, you cannot change this. What is the optimal seeding density and concentration of. Incucyte nuclight rapid red reagent for livecell nuclear.
The growth curve was plotted using the incucyte zoom software by analyzing phasecontrast or red object count for nuclight red. Incucyte zoom technical details incucyte zoom essen bioscience in incubator live cell imaging system for optimum environmental stability and reproducible imaging over extended timecourses. How to best statistically analyse results of an incucyte multidrug screen. Incucyte nuclight rapid red reagent for livecell nuclear labeling. Our nuclight bacmam and lentiviralbased labeling reagents enable expression of a nuclearrestricted green gfp or red mkate2 fluorescent protein in your choice of pri. This imaging system in combination with cellular dyes, stem cell specific stains and antibodies is an ideal solution for real time tracking of reprogramming, dissecting basic biology and screening for modulators of pluripotency maintenance and differentiation. Incucyte nuclight rapid red reagent for nuclear labeling. Each vial sufficient for 100200 live cell imaging cytotoxicity tests in 96well format. Peripheral blood mononuclear cells pbmcs were activated and labelled with incucyte cytolight rapid green dye and then added to tumor cells stably expressing a red fluorescent protein cytolight. It is compatible with the incucyte livecell analysis system and describes how incucyte nuclight rapid red can be used to fluorescently. In addition, this reagent can be multiplexed with incucyte annexin v green, caspase37 green, or cytotox green reagents for simultaneous readouts. Cells free fulltext cd44s assembles hyaluronan coat on. Mar 24, 2017 peripheral blood mononuclear cells pbmcs were activated and labelled with incucyte cytolight rapid green dye and then added to tumor cells stably expressing a red fluorescent protein cytolight. Incucyte cytolight rapid red reagent for live cell cytoplasmic labeling supplied as five vials, sufficient for 100 to tests.
Gabpadependent downregulation of dicer1 in follicular. Protect incucyte nuclight rapid red reagent from light at all times. Incucyte nuclight rapid red reagent for cell nuclear labeling. It is in an incubator so that cells can grow and be imaged in the normal plates that you would use for tissue culture. Cell culture media was supplemented with incucyte nuclight rapid red reagent for cell labelling essen bioscience and incucyte caspase. Using incucyte for automatic quantification of live cell analysis. Gain realtime, quantitative analysis of live cells with the incucyte s3 livecell analysis system by essen bioscience. Supplied as five vials, sufficient for 100 to tests. Addition of the incucyte nuclight rapid red reagent to normal healthy cells is nonperturbing to cell growth and morphology and provides homogenous. Experiment current method incucyte zoom assay description quality control and experiment optimization not tried before optimization of seeding conditions, cell labelling, media, assay. Squamous cell carcinomas scc of the lung lscc and head and neck hnscc are very prevalent with poor prognosis and limited treatment options. Add nuclight rapid red monitor proliferation of cells exact cell count 7.
Glutamine antagonism attenuates physical and cognitive. M, media was changed every 24 hours, and cells were imaged by the incucyte. Have your plate in the instrument at least 15 mins before starting first scan to allow condensation to settle. Livecell analysis handbook 2nd edition by sartorius. Using incucyte for automatic quantification of live cell. Incucyte cytotox red reagent for counting dead cells. If the incucyte instrument only exports a partial plate e. Shown here is a representative graph exported from the incucyte software. Mar 16, 2020 different concentrations of galactose. Immediately post addition, cells are labeled with the nuclight rapid red reagent. Incucyte nuclight rapid red reagent by essen bioscience.
This protocol provides an overview of the incucyte nuclight rapid cell labeling reagent methodology. Furthermore, the incucyte annexin v reagents can be combined with the incucyte confluence metric, incucyte caspase 37 reagent, incucyte nuclight nuclear labeling reagents or incucyte cytotox reagents for multiplexed measurements of apoptosis, cell proliferation or cytotoxicity in every assay well. Incucyte nuclight live cell labeling reagents enable efficient, nonperturbing, nuclear labeling of living cells and were used to label cell. The nuclei were stained with incucyte nuclight rapid red reagent essen biosciences, hertfordshire, uk and incucyte s3 2018b software essen biosciences, hertfordshire, uk was used to count the numbers of cells and the level of confluency.
Real time measurement of tumor cell death using incucyte. Oct 03, 2016 please use one of the following formats to cite this article in your essay, paper or report. Fluorescent celllabeling strategies for incucyte live. These dyes are well suited for identifying the location of cells within a po.
Cells free fulltext cd44s assembles hyaluronan coat. Incucyte cytolight rapid red reagent for live cell cytoplasmic. Incucyte nuclight live cell labeling reagents enable efficient, nonperturbing, nuclear labeling of living cells and were used to. These versatile labeling reagents have been validated. When stored as described the stock solutions will be stable for at least 12 months. Mutations in the mirna enzyme gene dicer1 have been reported in several endocrine malignancies and is associated with the rare tumourpredisposing dicer1. Incucyte nuclight reagents enable efficient, nonperturbing, nuclear labeling of living cells, using a range of different celllabelling strategies. Nuclight red reagents incucyte nuclight reagents enable efficient, nonperturbing, nuclear labeling of living cells, using a range of different celllabelling strategies. Incucyte s3 livecell analysis system from essen bioscience. Realtime and quantitative fluorescent livecell imaging. In the next experiment, hela nuclight red cells were treated with ssp in the presence of 5m incucyte caspase37 reagent and phasecontrast, red, and green images were collected every 2 hours in.
Fluorescent celllabeling strategies for incucyte livecell. Incucyte nuclight rapid red reagent essen bioscience. Incucyte cytotox green reagent for counting dead cells. Start with viable, living cells with incucyte s3 livecell analysis, cells are measured in situ in a physiologically relevant environment. Incucyte nuclight rapid red reagent for cell labeling.
Each vial sufficient for labelling labels 10100 million cells. The incucyte nuclight rapid red reagent for nuclear labeling is a cell permeable dna stain that specifically stains nuclei in cells and is ideally suited to the. Automated timelapse imaging of multiple positions with 4x 0. After 24 h, the cells were treated with varying doses 2100. Incucyte imaging for fluorescence measurements of a cell proliferation based on cell count in a ht1080 monoculture including the nuclight rapid red reagent b cell proliferation based on count in a coculture of ht1080 nuclight green and a549 nuclight red cells and c immune cell killing of skov3 nuclight red cells by activated pbmcs. Incucyte nuclight rapid nir reagent incucyte eshop. Following cell seeding, place plates at ambient temperature. These dyes are well suited for identifying the location of cells within a population and monitoring the dynamic interactions between cells in mixed cultures. Plates were transferred into the incucyte s3 live cell analysis systems and incubated at physiological condition 37 c, 5% co 2, over 72 h. The incucyte s3 is our flagship livecell analysis system, it supports the workflow and workload of larger laboratories the incucyte s3 livecell analysis system automatically acquires and analyzes images around the clock, enabling you to derive deeper and more physiologically relevant information about your cells, plus realtime kinetic data without ever removing your cells from the. Incucyte nuclight reagents cell proliferation essen bioscience. Essen bioscience launches the incucyte s3 livecell analysis platform enables realtime measurements of cell health, proliferation, movement and function while cells remain unperturbed in a. Incucyte zoom video tutorial and user guide the cell.
Essen bioscience launches the incucyte s3 livecell. Incucyte nuclight rapid red reagent qty 10 by essen bioscience. Incucyte nuclight lentivirus reagents provide homogenous expression of a nuclearrestricted gfp green fluorescent protein or mkate2 red fluorescent protein in your choice of primary, immortalized, dividing, or nondividing cells without altering cell function and with minimal toxicity. I have performed a drug screen consisting of 3drugs in different combinations against 3 separate cancer cell lines. The incucyte microscope has a 10x and 4x lens these have to be changed manually and it can take images of gfp, rfp etc. The strength of the incucyte is definitely the software package included.
Images were collected every 2 h, and proliferation was evaluated as number. Incucyte nuclight lentivirus reagents enable efficient, nonperturbing, nuclear. Incucyte nuclight rapid red cell labeling protocol. Nutraceutical effects of emblica officinalis in age.