Software Modules for the IncuCyte® ZOOM System - Structured Review
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A guided interface and purpose-built tools make the process of acquiring, viewing, analyzing and sharing images of 20016b cells easier than ever before using the Classic or vownload AI-driven Analysis. AI-driven Confluence Analysis provides a simple 2016bb for highly accurate segmentation of cells in phase contrast images, adapting to a wide range of incjcyte types and morphologies with minimal user input.
By viewing images of all downlosd in best download speed for zoom написано experiment at once, you can quickly assess treatment effects or identify outliers.
Dpwnload can also overlay metrics for rapid assessment and verification of image processing parameters. Then, generate presentation-ready images and movies with just a few clicks. Easily view images of all locations in an experiment at once to scan for trends and outliers, then zoom incucyte zoom 2016b download on wells of interest.
By providing purpose-built tools to answer your scientific questions, image processing and analysis is uncomplicated yet extremely powerful. Create analysis definitions once, then reapply to subsequent experiments to generate real-time подробнее на этой странице that enable decision-making.
Guided analysis interface ссылка на страницу even first-time users to convert images into insights using powerful AI-driven or classic segmentation for confluence measurements. Graphing tools enable review of trends and rapid generation нажмите для деталей publication- and presentation-ready graphs. Perform label-free cell counts incucyte zoom 2016b download subsequent cell-by-cell classification based on shape, size or fluorescence intensity to quantify dynamics changes in cell subsets within heterogenous living cell cultures.
Characterize the differentiation and maturation of organoid cultures in or well plates and assess treatment effects on organoid growth in well microplates. Analyze growth, viability or invasion of single spheroids in round- bottom multi-well format or measure multiple spheroid in flat bottom incucyte zoom 2016b download to detect changes of growth or viability. Enables label-free incucyte zoom 2016b download fluorescence analysis of neurite incucyte zoom 2016b download, maturation or disruption in each incucyte zoom 2016b download in or well plates.
Track zooom quantify incucyet and fluorescence labeled chemotaxis cell migration and invasion in microplate format. Assess complex network development and disruption qualitatively to study therapeutic interventions of vascular formation.
Gain insights into biological processes of cells in real time via non-perturbing incucyte zoom 2016b download analysis. Ideal if you use your Incucyte intensively as it keeps data on-line and at your fingertips for longer. The content of our website is always available in English and partly incicyte other languages.
Choose your preferred language and we will show you the content in that language, if available. Stress-free Image Acquisition Guided interface enables even first-time users to set up long-term, live-cell experiments Flexible scheduling tool allows incucyte zoom 2016b download users to run multiple applications in parallel.
Powerful, Purpose-built AI-driven Analysis Guided interface makes processing thousands of wells of data easier, driven by AI or Classic segmentation Graphing tools designed by biologists enable publication-ready graphs without the need for third-party software. Ordering Information. Software Страница Description Cat.
Add-on Software Modules. Classifier downllad trained using control wells. Literature and Documentation. Related Products.
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I can revoke my consent at any time with zkom for the future by sending an e-mail to unsubscribe sartorius. Request a Demo, Literature, or More Information. Language Preference. Please select your country so we can show you products that are available for you. 22016b counts and track changes in adherent cell morphology via label-free image segmentation and multivariate analysis of cell shape.
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The new PMC design is here! Learn more about navigating our updated article layout. The PMC legacy view will also be available for a limited time. Federal government websites often end in. The site is secure. Zlom it was hypothesized that a culture of ESCs was heterogeneous in the cell cycle, it was expected that increased S-G2-M-phases of the cell cycle would make an S-shaped curve parallel to the accumulation curve.
G1 delay by infrequent читать полностью change is incucyte zoom 2016b download mild stress, as it does not affect growth significantly when frequency is increased нажмите для продолжения 12 h. Comprehensive in vitro developmental toxicity DevTox assays are controversial, and the current standard is in vivo rodent gestational testing for the Extended One-Generation Incucyte zoom 2016b download Toxicity Study Beekhuijzen et al.
Previous ESC tests for cytotoxicity, cardiotoxicity Genschow et al. Many tests of pluripotent stem cells pre-differentiate these cells to other cell types, but the studies mentioned above immediately assess toxicant dose-dependent effects on stem cells cultured as stem cells. Cell-based HTS are an important part of drug discovery and ссылка на страницу in assessing the risk of drug and environmental compound zook Macarron et al. An important concept for DevTox in vitro HTS is that the cells used retain the developmental, molecular, and cellular strategies to respond to toxicant stress in the same way that they used to in the milieu of the early in vivo post-fertilization embryo /20736.txt were isolated from Rappolee et al.
Embryonic and placental incucyte zoom 2016b download stem cells ESCs and TSCs, respectively are isolated from the early embryo just prior to implantation into the uterus. In vitro, incucyte zoom 2016b download stem cell lineages do seem to emulate the strategies needed for the survival of the stressed embryo, in vivo. The early embryo at implantation has ESCs and TSCs, and both lineages enter 206b unique period of exponential growth as the two lineages arise, and this period lasts for about 10 days during mouse gestation and for 2—3 weeks in human gestation.
Exponential growth supports rapid embryonic expansion and sequential differentiation of the first few differentiated lineages from either ESCs or TSCs. The oncucyte differentiated lineages to arise are essential for the survival of the implanted embryo. In vivoif sufficient parenchymal function is produced, the embryo survives.
Stress from medical IVF увидеть больше toxicological sources should decrease stem cell growth and force first lineage differentiation to compensate for insufficient cell number and parenchymal function. Monitoring cell growth in every well of a high-throughput screen HTS is not possible by the naked incucyyte, and it is necessary to use an automated live imager or microplate reader to assay the cell number, nuclear number, or confluence in every download a zoom meeting of the HTS plate Boutros et al.
Growth reduction by dose-dependent stress is an important basis to integrate other downstream outcomes such as lineage imbalance or epigenetic deviation which are proportional to growth reduction Puscheck et al. However, the growth effects of stress are complicated as rapidly growing cells with zero stress dose may become contact-inhibited and confluent at Tfinal which is needed to optimize the detection of other developmental outcomes. The reliability of growth reduction assays incucyte zoom 2016b download marred by sownload confounding decrease in cell growth rates in lower stress-dosed wells that reach confluence earlier.
Hypothetically, FUCCI information regarding zoom app cell cycle progression would be diminished by stress on the first day of exposure, predicting a decrease in confluence and cell number or nuclear number /17822.txt the same stress incucyte zoom 2016b download dose at Tfinal.
Phthalates are esters of phthalic acid which are used in cosmetics, incucyte zoom 2016b download care products, and polyvinyl chloride PVC plastics Dutta et al. Since phthalates are not /28046.txt bonded into plastics, they enter the environment and then the body продолжить чтение absorption, ingestion, or inhalation.
Specifically, diethyl phthalate DEP is a low molecular weight LMW phthalate often used as a solvent in the manufacture of personal care products. DEP has previously been shown to increase neural markers at non-cytotoxic doses but slows mouse embryonic stem cell growth Yin et al. PFOA is a surfactant used in many industries, such as textiles and floor wax synthesis and in fire-fighting foam and sealants Lindstrom et al.
PFOA is a weak embryotoxic chemical often acting as an endocrine disruptor as defined by embryonic stem cells ESCs but can incucyte zoom 2016b download additive but not synergistic effects with BPA Zhou et al. This is relevant because real-world stressors are episodic, repeated simultaneously and synergistically, intrinsic, and environmental Awonuga et al. Traditionally each toxicant is assigned incucyte zoom 2016b download and incucyte zoom 2016b download adverse effect levels NOAEL incucyte zoom 2016b download LOAEL, respectively and half maximal toxic dose for each effect measured IC 50but other points of departure have been used more incucyte zoom 2016b download.
One such point of departure that we have also used is the benchmark dose lower confidence limitknown as the BMDL. We used the direness concept and dose-dependent kinetics of stressed stem cell responses reported by the live imager, to develop a separate predictive outcome for growth suppression.
The nadir is rapidly reversed into a green fluorescence peak by medium change. Here, we use toxicant dose-dependent suppression of this fed green peak as a взято отсюда indicator of future growth suppression, free of the confounding variable of contact inhibition of growth in low stress-dose responders that reach этим kahoot zoom app download моему at the earliest. ESC-qualified EmbryoMax fetal bovine serum, 0.
MEM non-essential amino acid solution, sorbitol, 2-mercaptoethanlol, and other zoo were obtained from Sigma St. Louis, MO. The osmolality of ESC media with and without addition of — mM sorbitol was determined previously Slater et al. The whole-well image was taken at both phase auto exposure and GFP channel ms.
The medium was changed every 24 h except for the experiment where the medium was changed every 12 h, but there was some variability in this schedule. Initial data analysis was conducted using acquisition software and was provided by IncuCyte Zoom Sartorius, version B This software analyzed the pictures to calculate the confluence and total green object integrated intensity of each well at each timepoint.
Total green object integrated intensity is the sum of incucyte zoom 2016b download intensities of the green objects. We calculated this stimulation incucte by, for each replicate, selecting the lowest nadir and single highest peak before and after medium change, respectively. The peak was chosen to be the highest value between T26—T36 or T50—T60, respectively the highest value within 10 h after taking the reading immediately after the average feeding time.
The trough was chosen to be the lowest point within 10 h preceding the chosen peak if the start of the peak was not easily seen. The analysis of time-dependent points of departure in this article is a simply the first significant adverse effect.
Benchmark dose modeling. We selected a benchmark response of one standard deviation of the control samples, as recommended by the USEPA Crump, We tested all models available in the default settings of BMDS, including linear, polynomial, exponential, and Hill models.
For DEP, we chose the frequentist Exponential degree 4 v1. Cultured pluripotent stem cells should grow exponentially after passage and be heterogeneous in the cell cycle. Stem cells in incucyte zoom 2016b download early mammalian embryo are not synchronized in the cell cycle, in contrast to the high degree of synchronization of pre-gastrulation stem cells in frogs and fruit flies Gilbert, People may use serum starvation and abnormal cell cycle synchronization to aid in preprogramming Chen et al.
Growth measured by confluence shows an S-shaped growth curve: In Посетить страницу источник 1A a live imager records confluence every 2 hours in five or six independent biological experiments for and downlkad feeding frequency, respectively. Standard ESC culture conditions require medium change every day to remove lactate from aerobic glycolytic.
Warburg metabolism was used by lncucyte cells to maintain good growth conditions and to incucyte zoom 2016b download the cells. ESCs found heterogeneously in the cell cycle seem like they would have an exponential increase in green cells with a line donload in shape and lower than the confluence line as shown in Figure 1A. Incucyte zoom 2016b download in Figure 1Bthe number of cells in the S-G2-M phase reaches a nadir in the last few hours prior to the h medium change and then increases in the hours following medium change.
It should be noted that increase продолжить green fluorescence in a single experiment varies from two—12 h after the nadir, although it is usually 6—8 h after the nadir. FUCCI ESCs accumulate in an S-shaped curve, but medium change reveals 201b6 cells accumulate in the green-negative state Продолжить чтение approaching a h interval feeding and then rapidly re-entering the green S-G2-M-phase after medium change.
The times for medium change may vary by 2 hours. Error zooom are s. It was hypothesized that insufficient medium change effects would incucyte zoom 2016b download ameliorated by more frequent medium changes. Two effects were expected; that the peak amplitude would decrease and the growth rate would increase with more frequent feedings.
In Figure 1Bit is shown that h feeding does decrease incucyte zoom 2016b download amplitude of the peak incucyte zoom 2016b download number of cells entering inxucyte S-G2-M phase after medium change, but in Figure 1A there was no significant increase in cell growth mediated by more frequent medium changes.
As shown in Figure zooncells are not heterogeneous in the cell cycle, and we /35332.txt the nadirs in приведу ссылку green object integrated intensity incucyte zoom 2016b download and after medium change by calculating the fold change between the peak divided by the nadir. Picking the nadir продолжение здесь peak for each replicate was performed to get the most accurate ratio as the timing of feeding and acquisition of green fluorescence may vary between experiments.
More detail of picking peaks is shown in the Statistics subsection of the Materials and Methods section. The nadirs from to h feeding frequencies were not significant, but the peaks for both frequencies were higher than the nadirs, and the h peak was significantly higher than incucyte zoom 2016b download h incucyte zoom 2016b download.
The stimulation index of h feeding frequency peak is greater than the download zoom free for pc h-feeding frequency peak. Продолжить чтение N as pro download zoom in Figure 1.
As по этому сообщению is reached and contact inhibition incucyte zoom 2016b download, the velocity of the lagging indicator of growth measurements decreases. Interestingly, the doubling rate is shortest, indicating fastest growth rate only on the first day, suggesting this would be a good period to assay for instantaneous changes in the growth rate caused by increase in incucyte zoom 2016b download stress doses.
Velocity of increase in confluence decreases with time, and doubling rates get slower with time as confluence and contact inhibition increases. A shows the fold change whereas B shows the doubling rate incucyte zoom 2016b download confluence from the stated time to 2 h. Five independent dowwnload experiments from h feeding frequency and six independent biological experiments for h feeding frequency with paired replicate wells in each experiment for each dose and stimulus were used for statistical analysis.
Confluence at T68 was used instead of T74 for datasets without T After LIF removal, ESCs slowly lose stemness and pluripotency-maintaining transcription factors, but the culture conditions here support similar time-dependent increases in confluence Figure 4A.
Similarly, instantaneous entry into the S-G2-M-phase showed a sharp increase from the unfed to fed states, though ND culture has a shallower increase than NS Figure 4B. A Cells were measured in time-lapse incucyte zoom 2016b download 2 hours for confluence in the live imager. Three highest PFOA doses suppress entry into the cell cycle, but only the two highest doses suppress the lagging phase accumulated growth measured by confluence. A Incicyte were measured in time-lapse every two hours for confluence in dowjload live imager.
As the cells grow for a longer period, the effect of the doses increases, causing the LOEL to be a lower dose. B Green cell cycle entry incucyte zoom 2016b download the S-G2-M-phase was measured simultaneously with confluence measurements in A and after feeding at day 1 and 2. In general, the green peak decreases as the dose of PFOA increases. NS 0 dose : N is described in Figure 1. PFOA: four biological experiments.
Analyses of the stimulation indices from the line graphs, now graphed as histograms, also show that PFOA suppresses accumulated growth as a lagging indicator measured by confluence Figure 6A and suppresses entry into the S-G2-M phase after medium change as a leading indicator Figure 2016n predicted downpoad ESC culture with LIF. In Figure 6ATfinal confluence means decrease with dose, but a second measure, the direness index, measures the first hour of departure of significantly decreased confluence compared with NS, 0 mM PFOA dose.
In general, the green fold change decreases as the dose of PFOA increases. The day 1 peak, occurring after the Aoom feeding, shows a clearer trend and incucyte zoom 2016b download less noisy than the day 2 peak which occurs after the T48 feeding.
In Figure 6Ca benchmark analysis of the point of departure was used to assess the dose of significant decrease compared with 0 mM sorbitol and normal stemness and such as Tfinal in Figure 6Ais accompanied by the direness significant point of departure for diminished confluence. As data of the confluences of the wells before the first feeding of the toxicant are not known, T0 was used 2 h after the first feeding for NS.
The direness points of departure from Figure 5A are shown in the day 3 bars. B The amount of green 0216b at the peak after feeding and the nadir before feeding is rationated.
Incucyte zoom 2016b download points of departure are shown in Figure 5A. All the models were deemed questionable.
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