Spark^® Cyto

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

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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

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.