Spark^®

LEARN – SELECT – MEASURE

A lot of research is devoted to the search for new compounds with bioactivity against bacteria. However, it is important to distinguish between antimicrobial activity and antibiofilm activity. Antimicrobial activity affects bacterial growth, and reduced growth can simultaneously result in a decrease in biofilm production. Antibiofilm activity only affects the formation of biofilms. Therefore, the use of antimicrobial compounds at concentrations that are not bactericidal against all susceptible bacteria often leads to the emergence of resistant strains. In contrast, antibiofilm compounds do not affect bacterial growth, and so do not generate the same opportunities for resistance to develop. This Application Note describes the SIMBA – SIMultaneous detection of antiMicrobial and anti-Biofilm Activity – method using the Spark multimode reader to generate OD600 growth curves for S. Infantis in a 96-well microplate over a 20- hour period, allowing the parallel determination of antimicrobial and antibiofilm activities of selected cold water fungal extracts.

Environmental pathogens are microorganisms that inhabit our surroundings, living outside of a human host for a large portion of their lifecycle. Many of these micro-organisms cause severe pathogenic conditions in plants and are often the reason behind significant crop failures that in turn lead to economic damage. Environmental pathogens can also cause diseases in humans and, in some cases, lead to severe infections that are difficult to treat. There is therefore a need for accurate methods to study these pathogens, and a common way to do that is through turbidimetric measurements. The aim of the current study was to investigate how bacterial growth is affected by temperature, using two different environmental pathogens – the rod-shaped Gram-negative Pseudomonas syringae and Erwinia amylovora.

Bioluminescence resonance energy transfer (BRET) is a well-established detection technology for the investigation of protein-protein interactions in living cells. The latest-generation BRET assay, NanoBRET™, employs an extremely bright NanoLuc® luciferase and a long-wavelength acceptor fluorophore, resulting in a markedly improved detection sensitivity and a higher dynamic range. This application note describes the implementation of NanoBRET technology on the Spark multimode reader, using the NanoBRET Target Engagement Intracellular HDAC Assay.

The novel ValitaTITER assay provides a very accurate and cost-effective solution for the detection and quantification of IgG and Fc-containing proteins directly in cell culture supernatants In this application note, we show the optimal protocol for performing IgG/Fc-protein quantification using ValitaTITER assay kits with the Spark and the Infinite 200 PRO multimode plate readers.

Temperature is a major factor affecting the rate and dynamics of biochemical reactions. The Spark® multimode reader’s ingenious Te-Cool™ module permits measurements even below ambient temperature, thereby limiting the effects of temperature increases in the laboratory on the dynamics of assays. It helps to stabilize temperature-dependent assays and makes the results more reproducible and reliable.

The Optimal Read function for fluorescence bottom measurements is designed to enable uniform well illumination during excitation and whole-well emission for 6-to 384-well plates. Performing multiple measurements on spatially separated spots arrayed across each well ensures that even heterogeneous cell layers can be effectively and reproducibly analyzed. This application note shows that the Optimal Read function produces the most reliable results for the analysis of adherent cell types, giving the highest well-to-well uniformity and best sensitivity for cell-based applications.

Cell counting, viability analysis and determination of size distribution are routinely performed by researchers across multiple disciplines, from standard cell passaging for downstream experiments to using insect cells infected with baculovirus to produce recombinant proteins, or determining the differentiation capacity of stem cells. Tecan’s Spark multimode reader has an integrated, bright field cell imaging module. This patent-pending system enables label-free cell counting, size distribution determination and trypan blue-based cell viability analysis with an easy-to-use, disposable Cell Chip™. In this study, the analysis of cell concentration, cell viability and size distribution for the determination of the optimal protein production rate in recombinant Sf21 cell cultures is presented.

HTRF is a time-resolved fluorescence resonance energy transfer (TR-FRET)-based assay system from Perkin Elmer cisbio, and is popular for the analysis of various molecular interactions and binding studies. HTRF assays place high sensitivity demands on multimode readers. The Spark offers a number of features to meet these demands and to ensure optimal assay performance, including Tecan’s unique Fusion Optics which combine the sensitivity of filters and the flexibility of monochromators (MCRs) in one instrument. In addition to entirely filter- or MCR-based measurements, a combination of the two can be used simultaneously within one measurement. Automated Z-position optimization and an integrated dichroic mirror that is perfectly suited to HTRF measurements further increase the Spark’s outstanding sensitivity. This application note describes the validation of the Spark reader, equipped with standard fluorescence optics, for HTRF measurements.

This application note describes the successful implementation of AlphaScreen on the Spark multimode reader, as well as the performance of the instrument with regards to measurement sensitivity, uniformity and typical read times.

This application note describes the implementation of the AlphaScreen technology on the Spark multimode reader and the detection of tyrosine kinase activity using the AlphaScreen P-Tyr-100 Assay Kit.

This application note describes the implementation of the AlphaLISA technology on the Spark multimode reader and its use for the detection of human immunoglobulin G (IgG) using the AlphaLISA IgG Assay Kit.

Tecan’s Spark multimode microplate reader with integrated cell imaging simplifies cell culture, featuring quality checks and signal normalization to cell confluence, thereby boosting automation of cell-based studies.

Keep control of crucial experimental conditions with the first air conditioning system for a multimode reader – enabling stable measurement chamber temperature down to 18°C.

Assaying cells in an appropriate window of time after a compound-­mediated cytotoxic event can be extremely challenging. This note describes the automation for kinetic monitoring of cytotoxicity and viability using three Promega assays for viability, cytotoxicity and apoptosis assessment with the Spark multimode reader.

Cell migration is a central process in the development and maintenance of multicellular organisms, with tissue formation during embryonic development, wound healing and immune responses all requiring the orchestrated movement and assembly of cells.

Tecan’s Spark multimode microplate reader was used for the real-time analysis of human stem cell proliferation and differentiation, using established assays for cell viability, proliferation and apoptosis assessment in a reliable and convenient manner.

Tecan’s Spark multimode microplate reader was used to identify metabolic changes during the differentiation of human mesenchymal stem cells and to assess and quantify cell viability, cytotoxicity and apoptosis, using several well-established and reliable cell-based assays developed by PromoCell.

For many cell biology applications – such as the identification and separation of high expression clones or knock-down candidates, and small-scale antibody or protein production – it is important to ensure that cell lines originates from a single cell, i.e. they are monoclonal. This can be a major bottleneck, especially for academic laboratories, as dedicated automated microscopy systems are both very expensive and complex to operate, meaning verification of monoclonality is often performed by manual microscopy.

Plate based titer quantification with PAIA titer assay and Tecan Spark® multimode reader.

Homogeneous Time-Resolved Fluorescence (HTRF), a time-resolved fluorescence resonance energy transfer (TR-FRET) assay system developed by Cisbio Bioassays, part of Revvity’s portfolio, is a highly reliable assay technology for studying molecular interactions. This application note summarizes the performance of the Spark reader with enhanced fluorescence optics for HTRF assays. All available optics configurations of the Spark’s Fusion Optics system meet the requirements for HTRF certification and are HTRF-compatible. Unless otherwise stated, the data presented was acquired using filter-based optics, as recommended for high performance HTRF measurements.

Fast and reliable nucleic acid quantification and purity checks are important daily routine applications for many laboratories, driven by the widespread use of next generation sequencing and other high throughput nucleic acid analysis techniques. Many methods require rapid, accurate analysis of small sample volumes to keep pace with downstream processes.This technical note shows the optimized Spark measurement settings, as well as comparative results, for the commonly used fluorescence-based DNA quantification assays evaluated in this study.

This technical note describes a method to perform UV[1]based protein quantification using the Spark multimode reader and the low volume NanoQuant Plate with the one-click SparkControl application. It shows comparative data for UV absorbance-based protein quantification in microplates and the gold standard for dye-based methods, the Pierce™ BCA assay. The study results can be used as a guideline to help scientists choose the right method for their specific application.

Evaporation has a huge impact on the performance of both biochemical and cell-based assays, changing reagent, component and metabolite concentrations to alter reaction kinetics and cell physiology. It is therefore crucial to keep the consistency throughout the whole plate to allow comparison of data between wells during life sciences experiments. This technical note highlights the benefits of using the Tecan’s Humidity Cassette in combination with the Spark. The Humidity Cassette can minimize the impact of edge effects in microplates under certain conditions. It provides better uniformity and delivers more reliable data for long-term studies.

Spark’s fluorescence module is available with a choice of filters, monochromators or Fusion Optics – a novel combination of both monochromators and filters. The Fusion Optics module combines the flexibility of monochromators with the sensitivity of filters in a single device, and even within individual measurements. In addition to this enormous flexibility, users can choose either a standard or an enhanced fluorescence module for their Spark multimode reader. The standard fluorescence module is tailored to basic applications, with an exceptionally good performance in the green wavelength range. The enhanced optics module is designed for higher throughput applications in 384- and 1,536-well plates, and offers better performance in the red and blue wavelength range, as well as for high throughput screening (HTS) assays, including TR-FRET and FP.This technical note describes the differences in the sensitivity-to-speed ratio of the standard and enhanced modules in 96- and 384-well plates for measurements using fluorescein (green) and Alexa Fluor® 555 (red).

Monitoring growth of prokaryotic cells and yeast is an important tool in life sciences, as well as in the food industry. This technical note demonstrates the Spark reader’s suitability for the generation of growth curves in the commonly used 24- and 96-well plate formats for three different, well-established bacterial strains, Escherichia coli, Listeria innocua and Staphylococcus capitis. It describes the established methods to generate OD 600 growth curves of all three bacterial strains in 24- and 96-well plates over a period of 8 hours. The influence of important instrument settings, such as shaking, are considered and discussed.

This technical note outlines the use of the Spark reader in combination with the ValitaTITER assay for the quantification of IgG. It offers a comparison of the use of the instrument’s standard and enhanced fluorescence optics, and discusses the optimization of flash number to produce high quality, reproducible data while minimizing read times.

This technical note outlines the use of the Spark reader in combination with the ValitaTITER assay for the quantification of IgG. It offers a comparison of the use of the instrument’s standard and enhanced fluorescence optics, and discusses the optimization of flash number to produce high quality, reproducible data while minimizing read times.

Spark-Stack is a compact, versatile and field-upgradeable microplate stacker for the Spark® multimode reader. It offers walkaway automation and enhances the productivity in the lab by helping to automate typical workflow steps – including plate loading, unloading and restacking of 6-1536-well microplates. Spark-Stack can be equipped with dark covers to protect light-sensitive assays inside the plate magazines, such as Alpha technology-based assays and fluorophore-transfected cells.

The one-click Live Viewer software application turns the Spark microplate reader into a microscope. It enables easy and straightforward visual quality checks of cells in microplate wells or in Tecan's proprietary Cell Chip™, allowing researchers to automatically acquire and instantly view images of their cell samples immediately before the measurement is started.

The Spark reader is equipped with Tecan's unique Fusion Optics, which combine the benefits of filters and MCRs in a single multimode reader. The Spark can be equipped with filters, Quad4 Monochromators or a combination of both using the ingenious Spark Fusion Optics.

The new Spark multimode microplate reader is equipped with patent-pending High Speed Monochromators (HSM) enabling highly accurate, reproducible and ultra-fast absorbance measurements. This unique optical system offers an enhanced measurement range from 200 to 1,000 nm, ensuring optimal performance across the full range, particularly for absorbance measurements in the deep UV range, such as A260/A230 nucleic acid purity checks. The Spark reader is also suitable for measurements of low volume samples in both absorbance and fluorescence modes using the patented NanoQuant plate.

With the Spark‘s luminescence module, users can perform high sensitivity flash and glow luminescence measurements. For optimized signal detection of very high and low signals within one experiment, optical density (OD) filters help to attenuate strong signals in the range from OD1 to OD3, thereby avoiding ‘over’ readings. This enables users to detect luminescence signals with a dynamic range of more than 9 orders of magnitude.Multicolor luminescence assays with up to five different luciferases, as well as luminescence spectral scanning (eg. of luminescent proteins) can be easily set - using an innovative filter-based approach, users can select any wavelength between 398 nm and 653 nm, with a step size of 15 nm and a 25 nm bandwidth.

This technical note describes the optimized test set-ups, instrument settings and statistical equations that were used to determine the maximum instrument sensitivity of the Spark multimode reader for flash and glow luminescence.

This technical note describes the optimized test set-ups, instrument settings and statistical equations that were used to determine the best possible detection limit for standard fluorophore label in fluorescence intensity top reading mode (FI top).

This technical note describes the optimized test set-ups, instrument settings and statistical equations that were used to determine the best possible detection limit of europium in time-resolved fluorescence intensity reading.

This technical note describes the optimized test set-ups, instrument settings and statistical equations that were used to determine the best possible detection limit for standard fluorophore label in FI bottom mode.

In this technical note, we demonstrate the superior performance of the Spark reader compared to two other multimode readers from different providers. The detection limit for the far red fluorescent dye Alexa Fluor 647 – which has a fluorescence spectrum comparable to Cy5 – was determined, operating each instrument in MCR mode.

This technical note describes the implementation of the DLR Assay System on Tecan’s next generation high-end multimode reader platform Spark. It can perform high sensitivity flash and glow luminescence measurements with a dynamic range of more than nine orders of magnitude. In addition, its enhanced luminescence module can measure multicolor assays with up to five different luciferases, as well as offering spectral scanning of luminescent substances.

This Technical Note describes the functions and benefits of the Spark multimode microplate reader and cell imaging module, as well as the SparkControl software and its dedicated Cell Counting and Cell Viability Application.

This technical note compares the performance of the Spark’s cell counting module with (i) manual cell counting using C-Chip™ disposable hemocytometers and a standard microscope, (ii) a comparably priced, imaging-based cell counting device (Device A) and (iii) a high performance cell imager with counting capabilities (Device B).

The Transcreener® ADP2 TR-FRET Red Assay is a competitive immunoassay for ADP with a far-red, time-resolved fluorescence-resonance-energy-transfer (TR-FRET) read-out. It is intended for the detection adenosine diphosphate (ADP) production by any kinase or ATPase, and has been developed specifically for high-throughput screening applications. The Spark was tested for its capacity to measure the Transcreener ADP2 TR-FRET Red Assay using the four different combinations of the Fusion Optics.

The Transcreener ADP2 FP assay is a competitive FP assay based on the detection of ADP and is therefore compatible with any enzyme class that produces ADP, including protein, lipid, and carbohydrate kinases, ATPases, DNA helicases, carboxylases and glutamine synthetase. This technical note describes the instrument settings for the Transcreener ADP2 FP assay on Tecan’s Spark multimode microplate reader.

The Transcreener ADP2 FI uses a simple fluorescence intensity read-out. It is a homogenous, competitive assay based on the detection of ADP with an extremely sensitive antibody against ADP. The Transcreener ADP2 FI assay is compatible with any enzyme class that produces ADP, including protein, lipid and carbohydrate kinases, ATPases, DNA helicases, carboxylases and glutamine synthetase. This technical note describes the instrument settings of Tecan’s Spark multimode reader for the Transcreener ADP2 FI assay.

The Host-Microbiota Interactions Lab at the Wellcome Sanger Institute uses high throughput anaerobic microbiology, genomic and metagenomic approaches to explore the relationship between humans and their microbiome – the community of microorganisms that live on and in us. HMIL has developed techniques to culture strict anaerobic gut bacteria at scale and built an extensive culture collection for phenotypic profiling. To phenotypically screen this culture collection, HMIL uses a Spark multimode reader equipped with an integrated stacker module – the Spark-Stack™ – for automated and high throughput plate processing, which is housed in a Whitley A135 HEPA Anaerobic Workstation. This allows the generation of bacterial growth curves cultured in multiple microplates under stringent anaerobic conditions, i.e. at 0 % O2 using a dedicated and pre-optimized gas mix (80 % N2 , 10 % CO2 and 10 % H2 ). Here, we describe accurate profiling of the growth kinetics of strict anaerobic gut bacteria.

Tecan’s Spark multimode microplate reader with integrated cell imaging simplifies cell culture, featuring quality checks and signal normalization to cell confluence, thereby boosting automation of cell-based studies.

Keep control of crucial experimental conditions with the first air conditioning system for a multimode reader – enabling stable measurement chamber temperature down to 18°C.

Cell-based assays are very popular in life sciences research, as they represent an in vitro system which offers results comparable to in vivo systems. However, this improved biological relevance is counterbalanced by the complexity of working with a living organism. Cell-based assays therefore require special precautions and care when planning and conducting experiments.

The Spark multimode microplate reader platform offers solutions to suit virtually any life science research and drug discovery applications. This system allows researchers to freely configure a reader to their needs and full upgradeability allows access to all techniques and features in the future.

Spark-Stack™ plate stacker for walkaway automation of Tecan Spark® readers Enhance the productivity of your lab with this compact, reliable and field-upgradeable plate stacker. Spark-Stack takes care of typical process steps, such as loading, unloading and re-stacking of non-lidded 6- to 1,536-well plates, and provides a throughput of up to 50 plates per run.