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Improving Fluorescence-Based Assay Performance Using Automated Digital Microscopy

Agilent Technologies
21 Apr 2015

One of the main limitations of fluorescence detection is background fluorescence, either from autofluorescence from endogenous cellular material, impurities from reagents added to the assay, unbound fluorescent conjugates or fluorogenic probes, or unreacted photoprotein. This study demonstrates three unique ways in which the incorporation of imaging, in addition to cellular analysis, allow for a more accurate analysis of the event taking place within the microplate well, and improve assay performance.

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Agilent BioTek Gen5 Software for Detection

Agilent Technologies

Agilent BioTek Gen5 software for Agilent BioTek multimode and single-mode microplate readers is an integrated tool for endpoint, kinetic, spectral scanning, and well area scanning. It controls all the functions of the plate reader and has powerful data analysis capabilities for a broad range of applications. Gen5 microplate reader software is also used to integrate BioTek plate readers to BioTek BioStack and other automated systems.

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Microplate Readers / DetectorsMicroplate readers are used to automate the detection and analysis of labeled or label-free components in microplates during assays or live-cell monitoring. Microplate readers are generally distinguished by their mode of detection. Types include absorbance, luminescence, fluorescence intensity, fluorescence polarization, TRF / FRET and multimode microplate readers. Microplate readers deliver a high throughput of samples by reading multiple wells simultaneously, with the 96-well format the most commonly used. As a result, microplate readers are often used in the drug discovery, bioassays, research and pharmaceutical industries for screening applications. Microplate loading can also be automated, with robotic microplate stackers to increase throughput. Find the best microplate readers in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.Digital MicroscopyDigital microscopy involves using digital cameras and sensors to capture high-resolution images of samples for analysis. It offers enhanced imaging capabilities compared to traditional optical microscopy and is widely used in biological and material science research. Explore digital microscopy systems in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.FluorescenceThe emission of fluorescence occurs when a photon of energy is supplied to a fluorescent chemical compound by an external source, causing it to become excited. Fluorescence can be detected and measured for different purposes using microplate readers, fluorescence microscopes, fluorescence scanners, and flow cytometers.AutomationAutomation in laboratories and manufacturing processes enhances efficiency, precision, and scalability by reducing the need for manual intervention. It plays a critical role in improving productivity, minimizing human error, and accelerating workflows in fields like diagnostics, drug development, and industrial testing. Automation technologies include robotic systems, automated liquid handlers, and process control systems that streamline complex tasks and ensure consistent, reproducible results. Explore our peer-reviewed product directory to discover the best automation solutions, compare options, read user reviews, and get prices directly from manufacturers.

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Improving Fluorescence-Based Assay Performance Using Automated Digital Microscopy