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Nature Methods Application Notes e-UPDATE: 09 October 2018

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09 October 2018 
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FEATURED APPLICATION NOTE
Airyscan detection in multiphoton microscopy: superresolution and improved signal-to-noise ratio beyond the confocal depth limit
https://www.zeiss.com/corporate/int/home.html >
The penetration depth of traditional confocal laser-scanning microscopy (LSM) systems is limited by light scattering. To avoid these limitations, multiphoton LSM uses a nonlinear fluorophore excitation process in combination with a non-descanned detection concept to greatly increase the penetration depth. However, in traditional multiphoton LSM, this increased depth necessitates a compromise on the achievable spatial resolution and signal-to-noise compared with that of confocal LSM. The novel Airsycan detection concept from ZEISS, used in combination with multiphoton excitation, overcomes these limitations and provides increased resolution and signal-to-noise with a 2–3× increase in penetration depth compared with that of traditional confocal LSM.
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TwistAmp® Liquid: a versatile amplification method to replace PCR
https://www.twistdx.co.uk/ >
Here we introduce TwistAmp® Liquid, a new PCR replacement format that makes RPA technology more amenable to a wide range of research applications. In contrast to PCR amplification, RPA takes minutes, rather than hours, and can be run with little to no equipment. TwistAmp® Liquid Basic and Basic RT kits can be used for applications requiring good fidelity, gel electrophoresis or solid phase detection. TwistAmp® Liquid exo and exo RT allow rapid real-time amplification and detection of targets without sacrificing sensitivity or specificity.
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High-content assay for morphological characterization of 3D neuronal networks in a microfluidic platform
https://www.moleculardevices.com/ >
Establishment of physiologically-relevant in vitro models is crucial to further understanding of the mechanisms of neurological diseases as well as targeted drug development. While iPSC-derived neurons show great promise for compound screening and disease modeling, use of three-dimensional (3D) cultures is emerging as a valid approach for neuronal cell based assay development. 3D cultures are recognized as more closely recapitulating aspects of the human tissues including the architecture, cell organization, as well as cell-cell and cell-matrix interactions1,2.
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Wheat germ cell-free protein expression enables studies on E3 ubiquitin ligases
http://www.cfsciences.com/eg/ >
With about 600 to 700 E3 ubiquitin ligases encoded within the human genome, it becomes a complex task to map out the substrate-E3 ligase interactions underlying the regulation of protein degradation in the cell. The wheat germ cell-free protein expression system from CFS, however, provides suitable means for preparing E3 ligases on a high throughout. This allowed different groups to do protein-protein interaction screening experiments and to identify binding partners for E3 ligases.
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Controlling the topography and biochemistry of cell culture substrates with PRIMO® photopatterning system
https://www.alveolelab.com >
In vivo, the cellular microenvironment has a crucial impact on the regulation of cell behavior and functions such as cellular differentiation, proliferation and migration. One of the challenges confronting cell biologists is to mimic this microenvironment in vivo to more efficiently study living cells and model diseases. Here we present PRIMO: a contactless and maskless UV projection system, which allows to control the topography and chemistry of in vivo microenvironments. We first show that PRIMO is a suitable tool to structure photosensitive resists and create molds on which elastomeric solutions (PDMS) can be polymerized. Then we show that the structured PDMS can be specifically functionalized with biomolecules using UV-light structured by PRIMO and a specific photo-initiator (PLPP™). Altogether, we demonstrate that PRIMO allows to tailor the cell microenvironment topography through microfabrication and biochemistry through micropatterning.
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Anti-idiotypic binders for Trastuzumab validated in regulatory bioanalysis assay in collaboration with Covance
https://www.avacta.com/ >
Regulatory bioanalysis of therapeutic proteins during drug development and clinical follow-up requires the use of critical reagents that can specifically identify and accurately quantify each biotherapeutic within patient samples. Antibodies currently represent the 'gold standard' of affinity reagents used in the regulated bioanalysis of therapeutic proteins. While traditional antibodies have been refined to the point where they are specific, sensitive and reasonably reliable, they can be limited by their development speed, complexity to produce on an industrial scale and lot-to-lot variation in assay performance.
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Improving biosensor assay development by determining sample quality with Tycho NT.6
https://nanotempertech.com/ >
Surface plasmon resonance (SPR) is an optical methodology widely used to detect and quantify molecular interactions. SPR is considered the gold standard for quantification of protein interactions, but SPR assay optimization can be technically challenging, time-consuming and costly. Tycho™ NT.6 quickly analyzes different conditions typically tested during optimization of an SPR assay. The Tycho™ NT.6 system is simple to use and enables researchers to make better-educated decisions in developing and optimizing their binding-interaction assays.
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