Cell biology

Cell separation has become instrumental in many areas of medical research over the last two decades, and Miltenyi Biotec’s MACS® Technology has become the gold standard technique, having been cited in over 20,000 publications. Combining superparamagnetic microparticles conjugated to specific antibodies with proprietary columns, this technology uses strong magnetic fields to separate specific cell types, allowing retention of both labeled and unlabeled cells for downstream analysis.

Since its launch at SLAS 2015 in February, the Spark® 10M has gained interest from across the life sciences community, with researchers around the world beginning to explore the advanced capabilities of this exciting new system for applications including DNA quantification, microbiology research and cell-based assays.

The Karlsruhe Institute of Technology has automated protocols for high throughput separation of cells for therapeutic applications on a Freedom EVO® 200 liquid handling platform. The process maintains cell viability and is more rapid and reproducible than manual methods, enabling processing of 96 samples in four hours.

The new Spark 10M multimode microplate reader is designed to offer greater flexibility and increased productivity for cell biology and genomics customers. From microbiology research and cell-based assays to rapid DNA quantification, the all-new platform delivers a combination of advanced capabilities and exceptional

A novel 3D cell culture platform is enabling scientists to generate microtissues that closely mimic the behavior of human tissues from virtually any cell type. This proprietary technology is easily automated, offering speed and versatility, combined with consistency and reproducibility, for high throughput applications.

Degenerative eye diseases affect millions of people around the world, causing progressive loss of vision and, in some cases, complete blindness. Researchers in Australia are using stem cell technologies to investigate ways of halting, or even reversing, the effects of common eye diseases.

The Karlsruhe Institute of Technology has automated protocols for high throughput separation of cells for therapeutic applications on a Freedom EVO® 200 liquid handling platform. The process maintains cell viability and is more rapid and reproducible than manual methods, enabling processing of 96 samples in four hours.

Tecan and TAP Biosystems have created an automated 3D cell culture screening platform by automating TAP’s RAFT™ cell culture technology on the Freedom EVO® workstation. This system uses in vivo-like concentrations of collagen matrix to ensure cell growth and proliferation in an environment closely resembling native tissue, with the added reliability and throughput of automated liquid handling.

Freiburg-based Oncotest GmbH specializes in preclinical pharmacological contract research, and has successfully automated 3D cell culture on a Freedom EVO® workstation using polystyrene-based Alvetex® Scaffold 96-well plates from Reinnervate

Tecan and InSphero have developed a novel solution for analyzing 3D cell cultures based on the Infinite® M200 PRO and InSphero’s 3D InSight™ microtissue technology. This solution is designed to facilitate the use of 3D cell culture formats in drug screening applications, helping to reduce the cost of early phase drug development.

The rising global problem of antimicrobial resistance has led to growing pressure on food producers to eliminate the use of prophylactic antibiotics as an additive in livestock feeds. To help combat the potential risk this poses to animal health, Agro BioSciences is harnessing the power of the microbiota present in the gut of chickens, pigs and cows to try and eliminate the growth of pathogens.

Researchers at the University of Montreal have benefitted from Tecan’s collaboration with SciRobotics to automate DNA assembly protocols for synthetic biology. Combining a Freedom EVO® 200 with a Pickolo™ Colony-Picker and various other components, the Systems Biology and Synthetic Biology Research Unit has built a streamlined cloning workflow with increased throughput and reliability.

Extended periods of walkaway operation are essential to the work carried out by Spanish energy producer Repsol, which relies on two Freedom EVO® platforms and Tecan’s 350 μl nested liquid handling disposable tips for its research into alternative energy sources.

Russia has been a fast-growing region for biotechnology and life sciences in recent years, with food and agriculture being key focus areas. Here is a snapshot from laboratories looking at the contrasting roles of microbiology in food production, both employing Tecan equipment in their workflows.

Laboratory automation and LIMS specialist Mejoran Lab Automation has developed a customized method for automated fungal colony picking on a Freedom EVO® 150 platform with an integrated SciRobotics Pickolo™ Colony-Picker. This innovative colony picking method produces viable microinocula of consistent quality for downstream assays, reducing the time needed to select mutants for further investigation from one week to one day.

The Life Sciences laboratory of the Central Research Institute at Ishihara Sangyo Kaisha Ltd has used Tecan’s Freedom EVO® 200 workstation to develop a successful automated protocol for efficient siRNA transfection, paving the way for efficient high throughput screening of small RNA libraries.

Automation with the Freedom EVO® system and SciRobotics’ Pickolo™ Colony-Picker has revolutionized the gene cloning and colony picking workflow at Evogene’s laboratory, allowing time-efficient and reliable tracking of cloning processes and enabling scientists to increase the yearly production of clones.

Tecan and amaxa GmbH (now Lonza) have successfully integrated the amaxa Nucleofector® 96-well Shuttle® System with the Tecan Freedom EVO® liquid handling workstation, allowing fully automated, high throughput transfection of primary cells and difficult-to-transfect cell lines, including non-dividing neurons and T cells, with siRNA, DNA and other substrates.

Discover tailor-made solutions for your cell biology workflow from cell maintenance and cell-based assays to life cell imaging.

Tecan has teamed up with pION Inc. to provide an automated solution for running high throughput drug permeability screening, using the pION Double-Sink™ parallel artificial membrane permeability assays (PAMPA™) and μSOL™ solubility assay. The flexibility of Tecan’s Freedom EVO® workstation and Infinite® M200 PRO microplate reader, combined with Pion’s Gut-Box™ microplate stirrer and PAMPA Evolution™ technology, provides a complete solution for drug candidate screening.

Tecan has created a range of automated ADME screening systems which maximize the efficiency and throughput of these processes. These systems are based on our liquid handling platform for automated cell permeability, compound characterization, in vitro drug metabolism and cell culturing procedures.

Discover a platform that speaks the language of your laboratory, and make your cell-based assays simpler, more efficient… and fluent.

Fraunhofer IME has adapted the Pluricyte Cardiomyocyte Differentiation Kit protocol for 96-well plates, using the Fluent Automation Workstation to create a screening platform for compound-related cardiotoxic effects.

Cytochrome P450 screening assay automated in a 384-well microplate format for lead generation using three different CYP enzymes.

The data presented in this application note underlines the suitability of the D300e Digital Dispenser for protein-based assays.

The results generated with the D300e Digital Dispenser are consistent with an expertly performed manual process, but with the benefit of significant time savings and the elimination of serial dilution.

Fully automated HDAC assays on the Fluent® Laboratory Automation Solution where triplicates of 10 serially diluted compounds per plate were tested for dose response to selective histone deacetylase inhibition.

The ability to carry out cell-based G-Protein coupled receptor (GPCR) assays in high throughput is a key part of the drug discovery process. Combining a Fluent® Laboratory Automation Solution configured for cell-based assays with the DiscoverX PathHunter® β-Arrestin human and ortholog GPCR assay kits gives users the power to run 384-well plate-based assays in high throughput.

Growth inhibition (GI50) of a set of eight compounds was tested on four different NCI-60 cell lines on a Fluent® Laboratory Automation Solution.

Easy automation of Cisbio’s EPIgeneous™ cell-based assays on a Fluent® Laboratory Automation Solution. In order to discover the optimal assay conditions, four different cell densities of three different cell lines were deployed to determine the level of H3K27Me3 and total H3 under different conditions.

Increasing sample numbers drive many laboratories to look for opportunities to automate and optimize their liquid handling processes. One of the dilemmas that laboratories face when configuring their automated liquid handling systems is the need to balance capacity and functionality against space and budget constraints.

The PAMPA Explorer Test System™ implemented on Tecan´s Infinite® M200 PRO multimode microplate reader comprises a quick, reliable system that accurately models both human jejunal permeability (GIT) and rodent blood-brain barrier (BBB) uptake of candidate drug molecules, based on measurements of permeability across artificial phospholipid membranes.

Screening of cytochrome P450 in combined dose-response and single concentration experiments in a 384 - well microplate format.

Automation of the fluorogenic Thrombin assay to identify coagulation factor lla inhibitors in a 384 - well microplate format

The Spark multimode microplate reader platform off ers solutions to suit virtually any life science research or drug discovery application. It allows researchers to freely confi gure the reader to their current needs, and is fully upgradable to access other techniques and features in the future.

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.

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.

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.

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.

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.

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.

Until now, it has only been possible to heat multimode readers using temperature control, not to set them at ambient temperature or lower. This can cause problems for temperature-sensitive experiments, such as enzymatic assays, affecting the reproducibility of results over time. In addition, the experimental precision can be decreased by the plate heating up during measurements, even over just a few minutes. Tecan’s Te-Cool™ module for the Spark® platform, the first air conditioning system for a multimode reader, addresses this problem. This unique device can cool the measurement chamber, enabling it to be set at any user-defined temperature from 18 to 42 °C, guaranteeing stable conditions every time, in every lab, across every season, anywhere in the world.

In cell culture applications, confluence assessment is an important quality control parameter that is commonly used to estimate the proportion of adherent cells on a growth surface as an indicator of the cell density inside a well or culture flask. Some cell lines behave differently regarding their growth rate or gene expression depending on the degree of confluence. Consistent determination of cell confluence and estimation of cell numbers are therefore important for reproducible assays and accurate interpretation of experimental data. However, visual cell imaging using microscopes is usually a time-consuming and laborious process, especially when multiple samples are being analyzed. A solution for this problem is automated cell confluence assessment, realized in the Spark® 20M multimode reader.

This application note describes the automated production and growth of single spheroids in ultra-low attachment (ULA) Corning 96-well spheroid microplates on a Fluent Automation Workstation, using a Multiple Channel Arm™ (MCA) for cell seeding and medium exchange.

Using automation to optimize 3d bloom microtissue conditions and drug efficacy screening in an ovarian cancer cell model.

CellSpring’s 3D Bloom microtissue platform is a new technology that shows promise for 3D cell culture. It can be used to make microtissues of virtually any cell type. The rapid assembly of cells within the 3D Bloom scaffold enables the generation of microtissues ready for testing in as little as 30 minutes, compared to hours or days using other methods.

TAP Biosystems (now part of Sartorius) has developed a 3D cell culture technique – RAFT™ (Real Architecture for 3D Tissue) – that allows researchers to culture their choice of cell type, including primary cells, in an in vivo-like environment. This application note documents the automated production of four 96-well RAFT cell culture plates in parallel, using the HCT-116 cancer cell line as an example for a typical tumoroid-forming cell line. It compares cellular viability and response to the anti-cancer compound doxorubicin in RAFT 3D cell cultures and classical 2D cell cultures.

Tecan and TAP Biosystems have developed an automated process for the preparation of RAFT™ cell cultures. Implementation on the Freedom EVO® system, with its modular automation capabilities, increases experimental versatility and analytical flexibility. In this application note, we present cell viability and proliferation data for primary neonatal human dermal fibroblasts obtained over an 11-day culture period after seeding.

Technological advances in the last couple of years have enabled 3D scaffolds to be engineered that offer a real proxy for the in vivo environment which can be easily and routinely used in a cell culture laboratory. The Alvetex® scaffold enables cells to develop natural, in vivo-like intercellular interactions, providing an ideal environment for real three-dimensional growth and nutrient exchange that is comparable to intra-capillary exchange in living tissues.

Combining Tecan’s Infinite® M200 PRO with InSphero’s organotypic microtissue tumor model provides a scalable system to assess drug sensitivity in complex 3D cell culture models. This allows long-term, tissue-based analyses – such as cell proliferation studies – offering reproducible measurements over time.

This application note demonstrates the potential advantages of automated stem cell culture in biopharmaceutical and cell-based therapy applications, where a consistent quality of cultured cells is essential.

Establishing protocols for automated harvesting of various cell lines from multi-well plates.

Researchers in the Department of Biochemical Engineering, University College London (UCL), UK, have automated stem cell culture on a custom-built Freedom EVO® 100 platform. Benefits compared to manual methods are higher consistency and reproducibility, better protection of cells from contamination, and improved operator safety.

This application note serves as a guide to adapt and establish harvesting protocols for various adherent cell lines growing in Corning® RoboFlask™ robot-friendly cell culture flasks using Tecan’s Freedom EVO® liquid handling platform equipped with a flask handling device (Flask Flipper).

TAP Biosystems (now part of Sartorius) and Tecan have worked together to develop an automated process that enables researchers to increase throughput, facilitate mixing procedures and enhance reliability while maintaining cell viability, reproducibility, experimental versatility and analytical flexibility.

To simplify the adoption of 3D techniques in drug discovery, this project investigated the automation of a hydrogel-based modular 3D cell culture system using a liquid handling robot. This approach was proven to be absolutely comparable to manually-produced hydrogels.

Oncotest maintains a unique repository of >300 tumor xenograft models representing all major histotypes as well as niche tumors. After evaluating many 3D cell culture platforms, Oncotest has chosen Alvetex® scaffold 96 because of its simplicity, low cost of adoption and flexibility to study tumor cell cytotoxicity, as well as mechanism of drug action at the translational and post-transcriptional levels.

The Institute for Research in Immunology and Cancer (IRIC) at the University of Montreal established a reliable, fully automated cloning workflow on a Freedom EVO® workstation. This system performs fully automated multi-fragment DNA assembly, customized plating in ANSI/SLAS-format plates, colony picking and PCR set-up – as well as agar plate preparation – allowing the group to greatly increase the throughput of its experiments.

A Freedom EVO® 150 workstation with a SciRobotics’ Pickolo™ Colony-Picker module proved the ideal solution to perform both fungal selection and the subsequent screening of different culture conditions. The Freedom EVO workstation can easily identify, select and pick fungal colonies based on user-selected criteria, and achieved 100 % microinoculum growth for subsequent experiments due to Pickolo’s unique picking procedures.

The Nucleofector® technology from Lonza has been specifically designed for difficult-to-transfect cell lines and primary cells, including stem cells, T lymphocytes and non-dividing neurons. This application note describes the integration of the Nucleofector into Tecan’s Freedom EVO® liquid handling platform to fully automate the transfection process.

Using primary cells for RNAi-based applications, such as target identification or validation, requires a highly efficient transfection technology in combination with a reliable and robust automation system. To accomplish these requirements, we integrated the Amaxa™ Nucleofector™ 96-well Shuttle® System into a Tecan Freedom EVO® cell transfection workstation.