Spark® Cyto

Plate reader with live cell imaging and real-time cytometry

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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Tab 01 / Overview
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More insights delivered in real-time, uncover new approaches with Spark Cyto

More cells analyzed

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 module

More parameters measured

Spark 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

More experimental control

Designed 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™

Spark Cyto capabilities*

Applications

  • Nuclei counting
  • Transfection efficiency
  • Cell viability
  • Apoptosis
  • Confluence assessment
  • Cell migration and wound healing
  • ELISAs
  • Low-volume DNA/RNA quantification
  • Nucleic acid labeling efficiency
  • Protein quantification
  • Reporter gene assays
  • HTRF®, DELFIA®, LanthaScreen®
  • Transcreener®
  • DLR®
  • BRET – including NanoBRET®
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Detection modes

  • Fluorescence imaging (blue, green, red, far red)
  • Bright field imaging
  • Digital phase contrast-imaging
  • Absorbance – incl. UV/VIS
  • Fluorescence top and bottom
  • Time resolved fluorescence (TRF)
  • Full spectral scanning capability for all measurement modes
  • FRET
  • TR-FRET
  • Fluorescence polarization (FP)
  • Luminescence – glow, flash, multicolor, scanning
  • AlphaScreen®, AlphaLISA® and AlphaPlex®
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Additional features

  • Reagent dispensers with heater and stirrer
  • Humidity Cassette
  • NanoQuant Plate™
  • QC tools for IQ/OQ services
  • Spark-Stack™ microplate stacker

* Capabilities depend on the Spark Cyto configuration.




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Tab 02 / Technology
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Optics engineered for speed and clarity

A dedicated optical set-up for live-cell cytometry in microplates

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.

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Schematic diagram of imager module. (1) LED for bright field; (2) microplate with sample; (3) objective; (4) multiband filter set; (5) LEDs and excitation filters for fluorescence; (6) Autofocus unit; (7) reflection mirror; (8) CMOS camera.

Built-in objective changer with 2x, 4x and 10x objectives.

Hassle-free selection of objectives and channels

Spark Cyto incorporates three objectives that can easily be selected on the fly in the SparkControl™ software:

  • 2x, numeric aperture =0.08
  • 4x, numeric aperture =0.13
  • 10x, numeric aperture =0.30

It also offers five LED channels to help meet the needs of your assay, without requiring manual user intervention.

  • Blue (381-450 nm)
  • Green (461-530 nm)
  • Red (543-611 nm)
  • Far red (626-800 nm)
  • Bright field and digital phase contrast
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Composite images without the pixel shift

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.

Autofocus enabled – stay focused on your research

Spark Cyto uses a patent-pending 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.

Live Viewer™

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.

Analyze the whole picture

Whole well view of a 96-well plate recorded with the 2x objective; HeLa cells were stained with calcein AM and propidium iodide, and analyzed with the cell viability application. The whole well image reveals an area within the well where the local cytoxicity is significantly higher, which can point to effects related to compound dispensing.

Whole well imaging of 96- and 384-well plates

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 patent-pending 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.

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Single 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.

One single image can tell the whole story

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 patent-pending 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.

  • 96-well plate – less than 12 minutes
  • 384-well plate – less than 45 minutes
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Imaging pattern for a whole well, 24-well recording using the 4x objective.

Whole-well imaging in other plate formats

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.

Tecan’s 96-well transparent cell culture plate. The Spark Cyto’s optics are developed and tested for these plates, guaranteeing the best possible image quality for whole well, 96-well analysis.

Optimized plates for optimal results

Tecan offers proprietary transparent tissue culture plates in 24-, 48-, 96- and 384-well formats. The Spark Cyto’s capabilities are specifically optimized for use with Tecan’s cell culture plates, assuring reduced optical artifacts and optimal results.




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Tab 03 / Applications
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Five predefined applications, even more possibilities

Intuitive image acquisition and analysis methods

Spark Cyto takes a user-friendly approach to the most common cytometry applications:

  • confluence
  • nucleic counting
  • transfection efficiency
  • cell viability
  • cell death

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.

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Image of viable and dead cells cpatured using the cell viability method with the object mask.

Whole well image from 96-well plate, acquired with the 2x objective; NNNHDF cells with confluence evaluation mask.

Confluence

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.

Nuclei counting

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.

Transfection efficiency

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.

Cell viability

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.

Cell death

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:

  • Blue objects – cell nuclei
  • Blue/red objects – necrotic cells (for live:dead cell ratio)
  • Blue/red/green objects – apoptotic cells (for apoptotic:necrotic cell ratio)
Show more

Transmission

Even more possibilities

Spark Cyto’s Method Editor offers two unique options for researchers looking to customize their assay:

  • User-defined protocols – set up specific imaging protocols for automated image acquisition and analysis
  • Imaging only – allowing acquisition and export of files to any third-party image analysis software, such as Image J or CellProfiler.

Real Time Experimental Control (REC™) – never miss a critical biological event!

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.

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Complete environmental control comes as standard

Spark Cyto comes equipped with a unique set of environmental features for controlled measurement conditions in your microplate:

  • Uniform temperature control (up to 42 °C)
  • Dynamic gas control (CO2 and O2)
  • Humidity control via Lid Lifter™ and Humidity Cassette

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.

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Humidity control for optimal evaporation protection

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 patent-pending Humidity Cassette is a cost-effective solution to minimize evaporation.

Lid Lifter

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.

Software designed for long-term studies

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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Spark Control™ – powerful control in an intuitive package

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.

Spark Control software is tailored to set up complex protocols and workflows for live cell analysis that include imaging-based analysis with standard detection technologies (RECTM – link to REC case). The easy and intuitive operation of the SparkControl’s workflow-based interface gives instant access to various applications, allowing the analysis of biological processes over time.

Image Analyzer™ – robust and accessible analysis tool

Images acquired with the Spark Cyto can be automatically processed with Image Analyzer, Tecan’s proprietary imaging software package. Image Analyzer offers you an array of customization options, making it easy to adjust and optimize your imaging parameters.

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.

Image Analyzer showing live HeLa cells stained with calcein AM and the respective object count and heat map for object size/RFU.




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Tab 05 / Configurations
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Configurations to meet your applications

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:

  • Lid Lifter™
  • Integrated gas control (CO2/O2)
  • Heating
  • LED-based autofocus
  • Objectives (2x, 4x and 10x)
  • 5-LED excitation, 4 color channels
  • Digital phase contrast
  • SparkControl™ software
  • Image Analyzer™ software
  • Instrument control unit

All four configurations can be optionally combined with additional features - see overview.




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Tab 06 / Accessories
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Injectors with heating and stirring

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.

Spark-Stack™ microplate stacker

Spark-Stack is a compact, automated plate stacker for integration with Spark readers. It provides reliable processing of up to 50 plates per run.

QC Tools and IQ/OQ services

Tecan’s quality control packages help you meet all regulatory requirements in an efficient, cost-effective way.

Read more

Humidity Cassette

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:

  • Reduction in evaporation during your assay
  • Use your existing plates – no need for expensive special plates
  • Better data quality – consistant results
  • Reduced edge effects – ideal for long-term kinetics
  • Stable conditions – better cell viability
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Tecan microplates

A collection of tissue-culture (TC) treated, sterile cell culture plates with individual lids for cell imaging and bottom-fluorescence measurements of cell-based applications. Performance assured with minimum background signal.

Read more

NanoQuant Plate™

The NanoQuant Plate allows parallel quantification and analysis of up to 16 samples in parallel, in a sample volume of as little as 2 µl. The unique design of the NanoQuant Plate, removes the need for calibration and guarantees consistent performance.




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Tab 07 / Literature
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Applications Notes

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

Technical Notes

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.

Whitepaper

Bringing time and cost advantages to labs…

Brochure

The Spark Cyto is a multimode reader platform equipped with a highly sophisticated fluorescence imaging module for real-time cytometry.

Read the Spark Cyto brochure

Typical performance values

Spark Cyto instrument specifications and typical performance values.

See the typical performance values for Spark Cyto

Spark Cyto is for research use only.