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Spark Cyto is a multi-mode plate reader with fluorescence imaging and cytometry capabilities, unlocking new possibilities for your cell-based research. By combining live cell imaging with industry-leading detection technologies, you now have the ability to unite qualitative and quantitative information into unique multi-parameter data sets.
Your cells don’t stay static when you leave the lab, so your research requires a dynamic instrument that ensures you never miss a key event. Spark Cyto works in real -time, using parallel data acquisition and analysis- to deliver meaningful insights faster than before.
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Silver winner of the Bioinformatics Life Science Industry Award 2019 in the category: |
The Spark Cyto can be an excellent support in any cell biology laboratory.
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Tab 01 / Overview
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Spark Cyto uniquely brings together top-of-the-range camera components with proprietary patent-pending technology to ensure that you can truly investigate your entire cell population. It gives you the ability to record the whole well area of a 96- or 384-well microplate with just one image, without tiling or distortion. This means that you never miss a cell when investigating the total cell population in a microplate well. Spark Cyto includes three magnification levels combined with four channels for fluorescence and bright field imaging, enabling high quality cell analysis for a wide variety of applications.
Learn more about the cell imaging moduleSpark Cyto is available in configurations, building on the foundation of the Spark multimode reader platform. It combines sophisticated imaging with proven multimode reader capabilities, allowing you to define new approaches to your research and obtain robust orthogonal data faster than ever before.
Learn more about the detection features of the Spark Cyto
Spark Cyto can be extended with an automated multi-plate cell incubator or integrated in Tecan’s fully automated workflow solutions.
Learn more how to scale up your researchDesigned to handle a broad range of common cytometry applications, Spark Cyto gives you a new level of experimental control without compromising on ease of use and convenience. Five predefined methods for common cytometry applications offer a straightforward approach to image acquisition and analysis, complemented by additional features such as ‘user-defined’ parameters and Real Time Experimental Control (REC™), making it possible for you to unlock new application possibilities
Learn more about SparkControl™ and Image Analyzer™* Capabilities depend on the Spark Cyto configuration.
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Tab 02 / Technology
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The Spark Cyto fluorescence imaging module is designed to deliver crisp images with minimal user intervention. Using three different objectives, five LEDs (bright field and fluorescence excitation), a multiband filter set, and a 12-bit CMOS camera, Spark Cyto eliminates pixel shift and delivers high quality images in a flash.
Show moreBuilt-in objective changer with 2x, 4x and 10x objectives.
Spark Cyto incorporates three objectives that can easily be selected on the fly in the SparkControl™ software:
It also offers five LED channels to help meet the needs of your assay, without requiring manual user intervention.
The optical system requires no movement of the optics or plate transport to image a well with any combination of the five available channels. This eliminates pixel shifts and results in remarkable image quality and acquisition speed, especially when using two or more channels.
Spark Cyto uses a patented LED-based autofocus system to deliver accurate images without compromising on scanning time. The autofocus system projects an extended grid pattern onto the sample surface, which minimizes the impact of potential distortions from isolated impurities, and then uses that as a guide to find the optimal focus for your samples. Simple, fast and effective autofocus that comes as standard on every instrument so that you never miss an image.
The Live Viewer software application enables the Spark Cyto to be used as an image cytometer, allowing cells to be monitored in real time. All optical channels and magnification levels are available in the Live Viewer, which can be operated from the software’s dashboard screen as a dedicated application, or from the Method Editor interface, to optimize parameters such as the focus offset or crosstalk between fluorescence channels before starting a measurement.
Spark Cyto allows you to image a whole-well in a 96- or 384-well plate with one single picture, giving you insight into every cell in every well. Using a proprietary patented wide field of view technology Spark Cyto obtains high quality whole-well images, without suffering from stitching artifacts or edge-to-edge image contrast. This gives you a complete picture of your cell population in less time, allowing you to drive your research in new directions.
Show moreSingle image of a whole well from a 96-well plate. No tiling or edge-to-edge optical distortion leads to superior results when analyzing cell populations.
Many imaging systems advertise the capability to acquire high quality images of an entire well, but most lack the ability to do this effectively. These systems create composite images, using tiling or stitching, which misrepresent the cell population and suffer from distortion due to fluid meniscus shadows on the well borders.
Spark Cyto captures the whole well (96- and 384-well plates) with a single image giving you a real picture of your research.
Spark Cyto uses a patented approach combining image acquisition with the 2x (96-well plates) or 4x (384-well plates) objective with a large camera chip and advanced imaging algorithms to give you accurate results with one image.
Imaging pattern for a whole well, 24-well recording using the 4x objective.
Primary cell lines tend to show better growth dynamics in 6- to 48-well plates. Spark Cyto allows whole well imaging for these applications, based on the automatic generation of composite images of multiple single image tiles.
Spark Cyto is optimized for Tecan’s cell culture plates and this combination guarantees the best possible image quality for your cell-based assays.
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Tab 03 / Applications
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Spark Cyto takes a user-friendly approach to the most common cytometry applications:
For all other applications simply select the ‘user-defined’ option to easily set up your own method in SparkControl™ or use the ‘image-only’ feature to export your files to a third-party image analysis software. This gives you complete flexibility in meeting your respective assay requirements.
Show moreWhole well image of a 96-well plate showing HeLa cells with bright field cell counting yellow overlay, acquired with the 4x objective.
This feature offers a fast and reliable way to count cells in bright field images directly, without the need of cellular dyes.
Using a deep learning algorithm, a yellow cross is overlaid on identified cells for easy visualization, and the total cell count is displayed for each well.
Whole well image from 96-well plate, acquired with the 2x objective; NNNHDF cells with confluence evaluation mask.
Use the bright field imaging channel to provide a quick overview of a well’s cell density. Cell confluence is calculated automatically by the software and displayed as a yellow overlay for easy visual confirmation. In addition, you can customize the roughness factor, a qualitative measurement, as a simple indicator of cell death.
Whole 384-well image acquired with the 4x objective; CHO cells with nuclei couting object mask.
Optimized for Hoechst 33342, this function provides an easy method for cell counting on the Spark Cyto, using any blue fluorescent dye with nuclear DNA binding capabilities.
Centered image of CHO cells cultured in a 96-well plate, acquired with the 4x objective, showing an overlay of the blue and green channels.
This application is designed to automatically determine the transfection rate for cells containing green fluorescent protein (GFP), a widely used reporter for gene expression, and counter-stained with Hoechst 33342 blue nuclei dye. The green and blue images are overlaid and analyzed to determine the resulting transfection efficiency in the cell population.
Centered image of HeLa cells cultured in a 24-well plate, acquired with the 10x objective, showing an overlay of the bright field, green and red channels.
Spark Cyto’s preset cell viability application relies on a common double staining approach to discriminate between live (green) and dead (red) cells in a population. Using two fluorescent dyes, suach as calcein AM (live cells) and propidium iodide (dead cells), you can image and analyze your population in minutes.
Image of A431 cells cultured in a 96-well plate, acquired with the 10x objective, showing an overlay of the blue, green and red channels.
The detection of, and discrimination between, apoptosis and necrosis can be accomplished by differential staining of markers characteristic of the relevant type of cell death, for example, with Hoechst 33342, propidium iodide and Annexin V-FITC.
Hoechst 33342 (blue) – nuclei stain
Propidium iodide (red) – necrotic cell stain
Annexin V-FITC / Alexa Fluor® 488 (green) – binds to the early apoptosis marker phosphatidylserine
Using a proprietary algorithm, the software can uniquely identify three object classes:
Spark Cyto’s easy to use Multi-Color application is ideal for counting and analyzing cells with multiple labels, using a fluorescence marker for the nucleus and up to two additional labels to automatically characterize your cells.
Transmission
Spark Cyto’s Method Editor offers two unique options for researchers looking to customize their assay:
REC grants you the ability to create new experimental workflows in your lab. Combining standard detection technologies, imaging capabilities and additional unique features, such as integrated humidity and environmental controls, REC unlocks new research possibilities.
REC uses all of these features to enable set-up of workflows that ensure you never miss a critical biological event – without chaining you to the bench.
Show moreSpark Cyto comes equipped with a unique set of environmental features for controlled measurement conditions in your microplate:
In combination, these features allow you to maintain a stable environment for your assays, effectively eliminating the risk temperature fluctuations or evaporation could pose to your results. Spark Cyto is the only instrument to put these features right at your fingertips.
All of these functions are easily programmed and controlled with SparkControl software.
Maintaining humidity levels of 95 % or higher is essential for unimpaired cell viability and growth, and miminizing evaporation is essential for maintaining consistent concentrations during long-term assays. Spark’s patented Humidity Cassette is a cost-effective solution to minimize evaporation.
Spark‘s integrated and patented lid lifting function creates new workflow possibilities, reducing the risk of sample contamination and helping to establish an ideal environment for long-term kinetic assays. Whether you want to dispense reagents without the need for manual intervention or maintain optimal environmental conditions without compromising evaporation protection, Spark Cyto is the only reader to offer this benefit.
Spark Control combines automation of long-term kinetic assays with advanced features such as kinetic conditioning and remote monitoring, providing a hands-off solution for complex experimental set-ups.
Cell-based kinetic experiments require specific actions to be performed once a certain signal threshold is reached. Multimode readers without conditional kinetics make these studies cumbersome and potentially error-prone. SparkControl’s kinetic conditioning feature allows you to program the system to automatically perform any required action as soon as a predefined signal is reached, enabling all the assay workflows you can think of.
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Tab 04 / Software
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The new SparkControl 3.2 software includes multiple features improving the overall user experience. For example, benefit from the new label-free cell counting functionality to simplify your cell-based assay workflow and save valuable time.
Already have a Spark? Please contact your local sales representative for a free upgrade to SparkControl 3.2.
SparkControl, best-in-class user-interface software, now includes a dedicated imaging stripe specifically for Spark Cyto. The imaging stripe can be combined with any existing stripe in SparkControl, making the creation of multiplex assays straightforward and effortless. The new stripe is streamlined to give the operator complete control of all the parameters associated with their experiment, without burdening them with superfluous information and wasting valuable time.
The powerful reader control software of the Spark Cyto, SparkControl includes a dedicated new stripe to set up and optimize automated cell imaging protocols. The imaging programming stripe can be combined with any other programming stripe of the conventional detection methods to allow easy multiplexing. The operator can select the objective and imaging channels of choice, define the field of view, control all paramaters associated with the measurement, or select one of the pre-defined methods for standard cell imaging applications. A quick link to the microscope-like Live Viewer mode gives immediate access to the image acquisition settings.
Stay connected with your experiment wherever you are. Use the Tecan Connect mobile app to monitor instrument status and alert you when user interactions are required.
Read moreImages acquired with the Spark Cyto can be automatically processed with Image Analyzer, Tecan’s proprietary imaging software package, offering easy data analysis for object segmentation, counting and gating.
Image Analyzer showing A431 cancer cells after induction of apoptosis, stained with three colors to allow determination of cell count (Hoechst), apoptosis (Annexin V-FITC / Alexa Fluor® 488), and necrosis (propidium iodide). Analysis Settings allows parameter settings for object segmentation and subsequent counting.
Images are saved in widely used jpg or tiff formats to make image analysis with alternative software products easy and seamless. In addition, Bio-Formats – an open source software plug-in that works with 150+ microscopy formats– is able to read Spark Cyto results, helping to simplify the use of a wide range of analysis tools.
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Tab 05 / Configurations
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Spark Cyto is available in four specialized configurations, which differ only in their detection modalities. This means that no matter the configuration of your Spark Cyto, you have a fully equipped system ready for real-time live cell imaging cytometry.
Every configuration of the Spark Cyto includes the following features for real-time cytometry, truly redefining what is standard for an imaging microplate reader:
All four configurations can be optionally combined with additional features - see overview.
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Tab 06 / Accessories
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Tecan’s Spark Motion concept enables walkaway automation for live cell experiments on up to 40 plates.
Integrated injectors permit automated reagent dispensing, ideal for high sensitivity applications such as flash luminescence. An attachable heater/stirrer unit reduces the risk of precipitation.
Built-in evaporation protection enhances live cell kinetic assays, minimizing edge effects for better uniformity and more reliable data.
A patented Humidity Cassette reduces evaporation in standard microplates, enabling long-term live kinetic studies without the need to switch to dedicated and costly microplate types – simply use your current, validated plates. Benefits include:
Stay connected with your experiment wherever you are. Use the Tecan Connect mobile app to monitor instrument status and alert you when user interactions are required.
Read moreTecan’s quality control packages help you meet all regulatory requirements in an efficient, cost-effective way.
Read more* Spark-Stack microplate stacker supports all read modes without imaging.
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Tab 07 / Webinars
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Combining live cell imaging with industry-leading detection technologies, you now have the ability to unite qualitative and quantitative information into unique multiparameter data sets.
These educational webinars explain how scientists advance their research with enabling cell analysis techniques.
Asst. Prof. Dannielle Engle, SALK institute
Pancreatic cancer is a deadly malignancy with few treatment options. This webinar highlights how dynamic measurements of confluence in patient-derived organoid models of pancreatic disease can predict treatment responses.
Dr. Christopher Wolff, FMP
If you are looking for a fast, uncomplicated and effective way of developing cytotoxicity cell painting assays, you won’t want to miss this webinar. Join Christopher Wolff from FMP Berlin and Christian Oberdanner from Tecan in an informative illustration of the precise and sensitive characterization of cytotoxicity based on multicolored imaging with Spark Cyto.
Dr. Julia Kirshner, zPREDICTA
Find out about the benefits of using 3D culture models in oncology drug discovery. This webinar will focus on the importance of the tumor micro-environment and tissue-associated extracellular matrix (ECM) for obtaining accurate drug response data, using the novel analytical capabilities of the Spark® Cyto multimode imaging plate reader.
Metabolic profiling of cancer cell line collections has become an invaluable tool in the study of disease etiology and drug modes of action, as well as for selecting personalized treatments. However, its scale is limited by time-consuming sampling and complex measurement procedures. Discover how the Institute of Molecular Systems Biology at ETH Zurich has developed a high throughput metabolomics platform to overcome these bottlenecks, using timelapse microscopy to combine dynamic phenotypic and molecular profiling at high throughput.
Dr. Marek Widera, Alexander Wilhelm
To decipher the pathology of COVID- 19, in vitro cell culture models that can physiologically mimic the viral replication cycle are required. This webinar introduces the A549-AT cell line, generated to meet this need and enable rapid and sensitive monitoring of SARS-CoV-2 replication and facilitating the characterization of viral variants.
Understanding when and how cells die following drug treatment is critical in the drug discovery and characterization process. However, traditional cell-based experiments that rely on endpoint data collection lack the details to answer these questions. Learn how to make more informed decisions on drug candidates with live-cell imaging and kinetic data using high-throughput amenable assays and instruments.
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Tab 08 / Literature
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The Spark Cyto is a multimode reader platform equipped with a highly sophisticated fluorescence imaging module for real-time cytometry.
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has become a global concern, due to its rapid spread. The study, published by the University of Frankfurt, describes the use of the Spark Cyto for the development of a cellular infection model that enables high throughput SARS-CoV-2 experiments and live cell imaging. The automated, non-invasive optical readout included assessment of confluency, roughness factor and fluorescence measurement. The data further highlights the use of the cell line for screening for antiviral compounds as well as for investigating the efficacy of neutralizing antibodies against different SARS-CoV-2 variants.
This application note systematically defines experimental parameters to enable live cell nuclear staining with minimal cytotoxicity during repeated exposure and continuous fluorescence imaging in the Spark Cyto, as well as robust automated cell segmentation with the SparkControl™ and Image Analyzer™ software.
The optimized procedure was used in a multiplexed, three-color assay to detect and distinguish early and late stages of apoptotic cell death, opening the door for dynamic long-term phenotype tracking.
Alzheimer’s disease (AD) and vascular dementia (VaD) are the two most common types of dementia, and their incidence is increasing year by year. They affect the health of the elderly, and cause a huge burden on society, with no effective treatments currently available.
In vitro research is the first step in drug evaluation, and cellular immunofluorescence can be used to more clearly define the role of drugs, provide more comprehensive and accurate information for downstream drug development.
This study uses the Spark Cyto – an innovative multimode reader with cell imaging capabilities – to evaluate the neuroprotective effects of miR-23b-3p (miRNA) and tilianin (compound) on stable transfected cell lines and human primary neurons. The results in this application note are based on a peer-reviewed article, published in the journal Oxidative Medicine and Cellular Longevity in August,2021, with the title: Tilianin Ameliorates Cognitive Dysfunction and Neuronal Damage in Rats with Vascular Dementia via p-CaMKII/ERK/CREB and ox-CaMKII-Dependent MAPK/NF-κB Pathways.
Scientific innovation is dependent on the power of observations, and the strength of the conclusions drawn from those observations. In in vitro biology, a majority of physiologically relevant outcomes adhere to dose- and exposure-dependent factors. Unfortunately, data collection for traditional cell-based experiments often occurs at arbitrary but convenient endpoints, and using inadequate or poorly informative tools. The resulting data typically lack the appropriate kinetic resolution to fully answer details of the cause-and-effect relationship. Because biological pathways and processes are already inherently complex, a more efficient screening paradigm is required, allowing scientists to examine the important parameters of ‘when’ and ‘how’ to better characterize a given response. This application note seeks to provide a practical demonstration of how real-time assays and a plate reader with bright field and fluorescence imaging functionality can work in unison to reveal cell- and compound-specific features of an apoptotic response.
This application note describes the outcome of an experiment comparing the Opera Phenix with the Spark Cyto for the detection of apoptotic cells. For this study, Hoechst 33342 was used as a ‘blue’ nucleic counter stain, with TMRM and YO-PRO-1 as the two secondary mask signals. Please note that findings presented here do not necessarily reflect the general performance difference between the two systems.
Workflow automation with the Spark Cyto multimode reader.
Automated monitoring of the reporter activity of hfob.1.19 cells following induction of osteogenic differentiation by temperature shift
While 3D cultures are superior to the 2D culture approaches in terms of physiological tissue organization and providing robust prediction of clinical outcomes, these techniques are often more expensive and time consuming to perform compared to the standard 2D protocols. Performing multiplexed assays with various readouts is therefore beneficial in order to gain the most information from each experiment. Here, we describe a multiplexed approach combining sequential imaging based LIVE/DEAD cell viability assays (Thermo Fisher Scientific) and CellTiter Glo luminescence analysis (Promega) using the Tecan Spark Cyto with r-Breast and r-mBreast models.
Analyzing the dose-response relationship using cell-based assays is essential to understanding a drug’s efficacy. However, fixing the appropriate dose range and time point is often challenging. The D300e Digital Dispenser enables accurate delivery of compounds at picoliter levels, spanning a wide dose range – for example, from 0.2 nM to 1 μM – in just a few minutes. This can be combined with the live-cell imaging and full cell incubation capabilities of the Spark Cyto multimode reader to monitor cell proliferation under standard culture conditions (37 ºC, 5 % CO2). This technical note uses the D300e and Spark Cyto to comprehensively capture an anti-cancer drug’s efficacy profile, tracking cell viability with nine doses at 25 time points using luminescence-based and bright field imaging technologies simultaneously.
This Technical Note briefly describes how to use Image Analyzer for the optimization of confluence assessments in bright field images, as well as for object segmentation and counting in fluorescence images.
This application note describes a semi-high throughput approach for the analysis of intracellular Ca2+ and cellular Ca2+ uptake using the Spark Cyto’s multicolor fluorescence imaging and the Image Analyzer™ software’s integrated Voronoi analysis function. Treatment with the calcium ionophore ionomycin was used to increase intracellular Ca2+ concentrations, and Fluo-4 was used in combination with the nuclear counterstain Hoechst 33342 for measurement quantification.
This technical note describes the capabilities of the Spark Cyto to normalize a cellular signal to the cell number, or cell mass, per well. This dramatically improves the overall data quality of the respective cell-based assay and, finally, leads to an improved reproducibility of cell-based experiments and more efficiency in the lab.
Fluorescence imaging using the spark cyto – acquisition in a snap using sparkcontrol™
Accurate and automated assessment of cell density using bright field and fluorescence imaging…
Automating visualization and quantification of protein expression in microplates…
Discriminating viable, apoptotic and necrotic cells.
Quantification of life: Dead less ratio with the Spark Cyto imaging cytometer.
Cell-based assays have a variety of practical uses within scientific research and are an integral component of many scientists’ workflows from testing tumor resistance for cancer research to helping identify novel therapeutic compounds in drug discovery.
The Spark Cyto is a multimode reader platform equipped with a highly sophisticated fluorescence imaging module for real-time cytometry.
Read the Spark Cyto brochureTecan’s Spark Motion concept enables walkaway automation for live cell experiments on up to 40 plates.
Novel cell models for rapid screening of SARS-CoV-2 infection studies
To decipher the pathology of COVID- 19, in vitro cell culture models that can physiologically mimic the viral replication cycle are required. This webinar introduces the A549-AT cell line, generated to meet this need and enable rapid and sensitive monitoring of SARS-CoV-2 replication and facilitating the characterization of viral variants.
Spark Cyto is for research use only.