Life Sciences


Creating Cell Repellent Surfaces in Microfl uidic Devices


AMSBIO’s Lipidure® - CM5206 has been used by a team of researchers from the Medical University of Vienna, to coat channels in their novel microfluidic devices, to prevent cell adhesion.


The aim of the research by the Medical University of Vienna was to create physiological-relevant in vitro tissue models that offer better predictability and the potential to improve drug screening outcomes in pre-clinical studies. Despite the advances of spheroid models in pharmaceutical screening applications, variations in spheroid size and consequential altered cell responses traditionally have led to non-reproducible and unpredictable results. Having developed a novel microfluidic multisize spheroid array - the researchers characterised it using liver, lung, colon, and skin cells as well as a triple-culture model of the blood-brain barrier to assess the effects of spheroid size on anticancer drug toxicity and compound penetration. The reproducible on-chip generation of 360 spheroids of five dimensions is demonstrated in a recently published paper.


Dr Mario Rothbauer, a Group Leader at the Medical University of Vienna, commented: “I’ve worked with anti-fouling surfaces for cell-based applications my entire career and so far, the Lipidure®-CM5206 has proved to be the most reliable and straight-forward approach for generation of cell repellent surfaces on microtitre plates, as well as more the complex culture environments of microfluidic, lab-on-a-chip and organ-on-a-chip systems. For our latest project we screened a panel of primary and cancer cell lines including lung, liver, gut, dermal fibroblasts and brain endothelial cells; Lipidure®-CM5206 did not let us down a single time throughout those many years.”


Lipidure®-CM5206 is a biocompatible and hydrophilic white copolymer made up of repeating units of 2-(methacryloyoxy) ethyl phosphorylcholine (MPC). Lipidure®-CM5206 is designed to mimic the cell membrane surface and its molecular structure is the key for its highly hydrophilic nature and extremely low toxicity.


Lipidure® coating of microtitre plates and microfluidic devices provides a superior low attachment surface for the production of state-of-the-art 3D cell culture. Formation of organoids, spheroids, tumorspheres, embryoid bodies and neurospheres using Lipidure® coated microtitre plates have been demonstrated for cell types including ES and iPS (human and mouse), NIH3T3, pre-adipocytes, HepG2 and other cancer cell lines as well as primary neuronal cells.


Read the Medical University of Vienna paper, ‘A Microfluidic Multisize Spheroid Array for Multiparametric Screening of Anticancer Drugs and Blood–Brain Barrier Transport Properties’: ilmt.co/PL/6E93 More information online: ilmt.co/PL/LqXm


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Fibre Lasers for 2-photon Microscopy and Optogenetics


A key technology in biological imaging in neuroscience, 2-photon fluorescence microscopy enables three-dimensional, non-invasive studies of the neuronal structure and activity on the submicron scale. The contrast mechanism in 2-photon microscopy in neuroscientific research is based on the excitation of green or red fluorescent proteins, so called GFPs and RFP, by two photons in the infrared spectral range.


To drive this nonlinear process and to resolve the neurons deep within the living brain, femtosecond lasers with clean temporal pulse shape and average output powers of >1W are an essential prerequisite.


Going beyond pure imaging, all-optical interrogation is a novel approach to understand how active patterns in neuronal activity drive behaviour. In such experiments the visualisation of neuronal activity by 2-photon imaging is combined with 2-photon optogenetics to stimulate individual neurons by photoactivation of channelrhodopsins within the cell.


With the need of exciting many neurons in parallel, the laser requirements and microscopy technology for 2-photon optogenetics are fundamentally different from 2-photon imaging. Typically, high-power multi-Watt lasers at 1030-1040nm with repetition rates in the 100kHz - 1MHz range are used in combination with a spatial light modulator (SLM) to excite 10s to 100s of neurons simultaneously.


To support and drive the research in neuroscience Toptica is proud to introduce their laser portfolio for 2-photon imaging of GFPs and RFPs, the FemtoFiber ultra 920 and FemtoFiber ultra 1050, and for 2-photon optogenetics of channelrhodopsins the FemtoFiber vario 1030 HP.


All lasers are fully matched to the requirements in neuroscientific research and besides being fully turn-key, quiet, and compact, they are offering integrated dispersion pre-compensation (GDD) and integrated power control (AOM) to simplify operation and to allow the scientist to focus on their research.


More information online: ilmt.co/PL/drWX 57098pr@reply-direct.com


Exploring the Identities and Modifi cations of Proteins


Researchers from the University of Oxford, in collaboration with the University of Liverpool’s Centre for Proteome Research and the Wellcome Sanger Institute have been awarded £5.5 million by the BBSRC in support of a project analysing individual proteins to decipher the complexity of bacterial communication. Over the next five years, the multi-disciplinary team aims to develop and apply a novel approach for identifying proteins and their common modifications, such as phosphorylation, which can drastically alter a protein’s function. These modifications are difficult to detect with existing technology meaning they remain largely hidden.


The project will leverage three technologies developed by members of the team; nanopore, electrometry and mass photometry, which are already used individually to extract information about biomolecules, including their mass and electric charge. Studying the role of phosphorylation in individual bacteria will enable improved understanding of microbial life, helping to better combat infection and antimicrobial resistance.


Project lead Professor Justin Benesch, from the University of Oxford’s Department of Chemistry, said: “Proteins carry out the processes of life, yet harbour much complexity that current technologies cannot detect. Our approach should reveal much of this and we really look forward to exploring what we will uncover.”


Professor Claire Eyers, Director of the Centre for Proteome Research, said: “This is a fantastic opportunity that will allow us to explore the multitude of differentially modified protein species that contribute to regulating functional responses, not only in pathogenic bacteria, but to all forms of life.”


The project was awarded a Strategic Longer and Larger (sLoLa) grant by the BBSRC. The sLoLa programme is designed to support frontier research that will address significant fundamental bioscience questions and improve our understanding of the fundamental ‘rules of life’. This project was one of just four chosen for funding.


More information online: ilmt.co/PL/ZZ6V 57306pr@reply-direct.com


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