Powerful and most sensitive HTS plate reader
The PHERAstar® FSX is our latest, most sensitive multi-mode reader for high-throughput screening, combining this sensitivity with fast read times. Thanks to the unique Optic Module system, Simultaneous Dual Emission, UV/vis spectrometer, and dedicated AlphaScreen® and TRF lasers, the PHERAstar FSX will perform your assay with ease and speed, even in 3456-well microplates.
No longer worry about installing filters and dichroics: the PHERAstar FSX comes with easy-to-handle, assay-optimized Optic Modules which contain all application-specific filters, mirrors, dichroics and/or polarizers, and are automatically recognized by the plate reader. In addition, the PHERAstar FSX is equipped with four matched and assay-optimized photomultiplier tubes (PMTs) which are automatically selected for the relevant detection mode. You can also measure assays with two emission wavelengths at the same time thanks to the Simultaneous Dual Emission.
For HTS automation purposes, the PHERAstar FSX offers improved robotic integration capabilities and multi-user control.
- The most sensitive reader in fluorescence intensity and polarization
- The only plate reader with 5 assay-dedicated detectors
- Simultaneous Dual Emission, including for Alpha Technology
- 9 decades luminescence dynamic range
- Dedicated AlphaScreen®/AlphaLISA®/AlphaPlex™ laser
- New generation TRF-laser
- Top and bottom focal height adjustment
- High-precision injectors with simultaneous reagent injection and detection
- High-speed absorbance measurements
Fluorescence Polarization based assay for rapid, precise, high-throughput measurement of IgG & Fc containing derivativesDr. Carolanne Doherty, Valitacell, NIBRT, Fosters Avenue, Blackrock, Dublin, Ireland, 11/2017
The accurate, rapid and high-throughput measurement of IgG is essential in the development and manufacture of most therapeutic antibodies. Monoclonal antibodies are becoming increasingly dominant in biopharmaceuticals, where a vast number of samples must be screened for the development of each potential therapeutic.
Here we report the development of a novel, rapid, and simple fluorescence polarization based assay for high-throughput titer measurement of IgG and Fc-containing derivatives. It uses fluorescently labeled protein G that binds IgG. Upon binding, the depolarization of emitted light decreases, this can be detected to report on the presence of IgG. The CLARIOstar®, PHERAstar® and POLARstar® exhibit excellent assay quality when used with the ValitaTITER assay, which enables a high-throughput, simple, precise method for quantification of IgG. Moreover, the assay can be performed using sample straight cell culture supernatant, which means there are no complex sample preparation steps.
Calcium retention capacity assay evaluates inhibition of mitochondrial permeability transition poreM. Awais , D. Latawiec , R. Sutton, Liverpool Pancreatitis Research Group, Institute of Translational Medicine, University of Liverpool, UK, 11/2017
Mitochondrial dysfunction is central to the pathogenesis of acute pancreatitis, ischemia-reperfusion injury of the heart, brain and kidney, muscular dystrophies and neurodegeneration. Mitochondrial dysfunction is the result of a sudden increase in permeability of the inner mitochondrial membrane (IMM), via persistent opening of a multi-protein channel known as the mitochondrial permeability transition pore (MPTP). This is followed by uncontrolled proton flow across the IMM and unregulated flux of water, ions and small solutes into and out of the mitochondrial matrix. This results in rupture of the outer mitochondrial membrane (OMM) and eventually cell death by necrosis. Therefore, MPTP is an attractive target for cell death prevention in a host of disease states.
The calcium retention capacity assay challenges isolated mitochondria with spikes of calcium ions. Upon opening of the MPTP, Ca2+ leaks into the assay buffer and increases fluorescence of the membrane-impermeable CalciumGreen™ dye. The Omega multi-mode plate reader has proven excellent robustness for performing the multiple injections as well as reliable fluorescent detection of the assay.
Protein-induced fluorescence enhancement detects protein-nucleic acid interactions in microplatesFulneček J. , Valuchová S. , Petrov A.P. , Tripsianes K. , Říha K., Research group Molecular Biology of Plants, Central European Institute of Technology, Masaryk University, Brno, Czech Republic, 10/2017
Various biological processes depend on the interaction of proteins with nucleic acids. These comprise transcriptional regulation, DNA replication and DNA repair. Conventional methods to study the interaction are either laborious (EMSA, ChIP) or limited to specific targets (EMSA, FP, ChIP).
A novel assay employs the Cy3 fluorophore which increases its fluorescence when being in proximity to proteins: Cy3-labeled oligonucleotides are immobilized on a microplate bottom and the basal fluorescence is measured on a microplate reader. If a protein is added to the microplate well that binds the oligonucleotide, the fluorescence of the fluorophore will increase. This increase can be measured and reports on protein-nucleic acid binding.
This simple and cost-effective assay detects sequence and structure specificities as well as binding constants of nucleic acid:protein liaisions. The change in fluorescence intensity is captured on a huge surface to provide stable measurements. This has been enabled by the well-scan function and high sensitivity of the FLUOstar® Omega.
Quantifying double-stranded DNA with fluorescent dyes: Qubit on BMG LABTECH instrumentsAndrea Krumm , Mark Gröne , Franka Maurer, BMG LABTECH GmbH, Ortenberg, Germany, 07/2017
The quantification of DNA is a standard laboratory activity and pivotal for subsequent applications such as next generation sequencing (NGS). The widely used Qubit™ nucleic acid staining dyes are analyzed on a Qubit™ fluorometer. It measures fluorescence intensity directly in sample preparation tubes and automatically calculates the DNA-concentration based on two standards measured in parallel. This provides an easy, quick and intuitive way to quantitate dsDNA. However, throughput is limited using the Qubit™ fluorometer as only one sample can be analyzed at a time.
This application not presents how the Qubit® dsDNA HS fluorophore can be used to quantitate dsDNA on a BMG LABTECH microplate reader. The Qubit kit read on the FLUOstar® Omega, CLARIOstar® and PHERAstar® resulted in a high linearity (R2 > 0.9999) and quantification of DNA dilutions of 50 pg/μl-100 ng/μl. Detecting Qubit chemistry on BMG LABTECH microplate readers increases throughput while keeping the simplicity of the quantification method.
Screening for inhibitors of the ubiquitination regulator and anticancer target CSN5 using a HTS fluorescence polarization methodU . Hassiepen , M.D. Jones , R. Assenberg , I. Bechtold , C. Logel , M. Renatus , B. Martoglio, Novartis Pharma AG , Switzerland, 04/2017
Cullin-RING E3 ubiquitin ligases (CRLs) are activated by neddylation and inactivated by de-neddylation that is accomplished by the COP9 signalosome (CSN). Trapping CRLs in their inactive state is reported to elevate tumor suppressors. Therefore, inhibition of CSN is a potential modality for cancer treatment.
Here, we show the development of a high-throughput de-neddylation assay for a CRL substrate that was used to find inhibitors of CSN. The fluorescence polarization assay was optimized using a fluorophore with comparatively long fluorescence lifetime.
Performing the assay in a 384 well plate and reading it on a PHERAstar® microplate reader provided an assay window of 130 mP. The reader's capability of simultaneous detection of the two polarization channels accelerated the measurement and enabled kinetic reads. Hence, the assay is applicable for both CSN inhibitor finding and characterization.