细胞生物学

Access to human pluripotent embryonic stem cells is enabling Genea Biocells to pioneer novel therapies to treat a number of neuromuscular diseases. Drawing on almost 30 years of research heritage, the company is using its expertise to model spinal muscular atrophy and facioscapulohumeral muscular dystrophy to identify potential therapies.

The Monash Antibody Technologies Facility (MATF) in Victoria, Australia, has added a new dimension to its high throughput service, offering screening for antibody functionality. This time-consuming phase is crucial for many projects, and can now be outsourced to MATF, where comprehensive and flexible automation completes testing in a fraction of the time.

The new Spark 20M multimode microplate reader offers tailor-made solutions to suit virtually any drug discovery or advanced life science research application. This freely configurable system gives researchers access to new techniques and features intended to enhance and streamline biochemical and cell-based workflows.

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.

Scientists in the ADME Screening, Enzymology and Automation Group at Roche, Switzerland, have successfully integrated a mosquito® nanoliter liquid handling instrument with a Freedom EVO® platform, enabling accurate, precise pipetting at very low volumes.

Scientists working in the Screening Unit at the Institute for Molecular Pharmacology in Germany are finding the low volume pipetting capability of the MultiChannel Arm™ 384 beneficial for the investigation of siRNAs and small molecule compound screening.

Tecan and pION Inc. have combined pION’s PAMPA Explorer Test System™ with Tecan’s microplate readers for drug permeability screening using the Double-Sink™ parallel artificial membrane permeability assay (PAMPA™).

The German army has automated the determination of AChE activity in whole blood. The determination of cholinesterase status is primarily intended for analyzing patients poisoned with organophosphates (accidental or suicidal poisoning), but it can also be applied after the use of nerve agents in military conflicts or terrorist attacks, as well as in the context of occupational medical examinations.

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.

自从2月份在SLAS 2015推出以来，Spark®10M已经在整个生命科学界获得了兴趣，全球研究人员开始探索这种令人兴奋的新系统的先进功能，包括DNA定量，微生物学研究和基于细胞测定。

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.

新的Spark 10M多功能酶标仪旨在为细胞生物学和基因组学客户提供更大的灵活性和更高的生产力。 从微生物学研究和基于细胞的测定到快速的DNA定量，全新的平台提供高级功能的组合并且非常易于使用，以简化您的日常工作。

一种新颖的3D细胞培养平台使科学家能够产生与几乎任何细胞类型的人类组织行为密切相似的微量组织。这种专有技术易于自动化，为高通量应用提供高速度,多功能性，一致性和可重复性。

退行性眼病对世界各地的数百万人造成影响，导致视力逐渐丧失，在某些情况下，甚至完全失明。澳大利亚的研究人员正在使用干细胞技术来研究如何阻止或甚至扭转常见眼睛疾病的影响。

Karlsruhe理工学院拥有在FreedomEVO®200液体处理平台上用于治疗应用的高通量分离细胞的自动化方案。该过程保持细胞活力，并且比手动方法更快速和可重复，使得能够在四小时内处理96个样品。

Tecan和TAP Biosystems通过在FreedomEVO®工作站上自动化TAP的RAFT™细胞培养技术，创建了一个自动化的3D细胞培养筛选平台。该系统使用类似于体内的胶原基质浓度来确保在与天然组织非常相似的环境中的细胞生长和增殖，并增加了自动化液体处理的可靠性和产量。

基于Freiburg的Oncotest有限公司专门从事临床前药理优选法研究，并在FreedomEVO®工作站上成功自动化3D细胞培养，使用了Reinnervate的聚苯乙烯基的Alvetex®Scaffold 96孔板

Tecan和InSphero开发了一种基于Infinite®M200 PRO和InSphero 3D InSight™微量组织技术以分析3D细胞培养的新颖解决方案。该解决方案旨在促进在药物筛选应用中使用3D细胞培养形式，有助于降低早期药物开发的成本。

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.

This white paper outlines how the Fluent® Automation Workstation was used to automate cell culture for beating, hiPSC-derived cardiomyocytes. It also describes the use of these cultured cells to screen >3,500 small molecules in a chemically induced hypertrophy disease model, using a validated phenotypic assay.

This application note describes the successful automation of cytotoxicity endpoint assays using mES-D3 and NIH 3T3 cells on the Fluent® Automation Workstation. Researchers at Fraunhofer ITMP ScreeningPort have combined automated cell culture maintenance and homogenous cell viability assays with high content imaging and analysis to optimize the EST and develop a robust, high throughput screening platform to predict negative effects on embryonic development in vivo.

This application note describes the successful automation of an mES-D3 differentiation assay, including embryoid body formation on the Fluent Automation Workstation. In combination with the automated workflow for compound toxicity assessment in mES-D3 and NIH3T3 cells, researchers at Fraunhofer ITMP ScreeningPort have optimized the EST workflow and developed a robust screening platform to predict negative effects on embryo development in vivo.

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.

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.

Streamlining 3D cell culture-based drug response profiling with the Fluent® Automation Workstation, D300 Digital Dispenser, and Spark® Cyto multimode reader.

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

Easy automation of a biochemical assay on the Fluent® Laboratory Automation Solution to identify coagulation factor IIa inhibitors in a 384-well plate 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.

There has been growing interest in the use of in vitro three-dimensional (3D) cell culture in cancer research and tissue biology over the last couple of decades. 3D spheroids are more physiologically relevant compared to conventional 2D cell culture, making them more suitable for in vitro efficacy testing and analysis of drug penetration, bridging the gap between traditional monolayer cultures and animal testing. Furthermore, 3D cell models are promising tools in personalized medicine, as tumor spheroids can be obtained from most tumors, and can be used to screen drugs to find effective patient-specific therapies. Alternatively, healthy multicellular spheroids are potential organ building blocks to restore lost organ functions in regenerative medicine applications.

Automated cell seeding, medium exchange and incubation of 3t3-l1 cells using the fluent automation workstation and thermo scientific’s cytomat™ 2 c450-lin automated incubator

This application note compares accuracy and reproducibility between manual and automated seeding of adherent cells in 96-well microplates using the Fluent Automation Workstation. We show how the Fluent platform can successfully increase productivity while achieving high quality results.

This application note describes optimized protocols for the maintenance and reprogramming of patient-derived fibroblast cells into induced pluripotent stem cells (iPSCs) and their differentiation into retinal cells using the Freedom EVO® platform.

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.

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.