High-throughput fluorescent colony formation assay

7 Jul 2021

The colony formation assay evaluates the proliferative capacity of a single cell. Conventional analysis of this assay involves scoring and quantifying colonies in each well of a multi-well format manually by eye, limiting its throughput capabilities.

In this application note, BioTek Instruments presents an automated high throughput method for conducting a colony formation assay in a 96-well microplate using fluorescence microscopy with the Cytation™ 5 wide field of view. This fluorescence-based method is a powerful application of a classic assay, with a streamlined analytical process.

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MicroplatesMicroplates are multi-well plates used to increase the throughput of biological investigations. The number of wells microplates contain can range from 6 to 3243 wells, with the 96-well format being the most commonly used. Microplates can come tailored for a range of applications including cell culture, PCR, filtration, storage, non-binding surface, protein crystallization, as well as pre-coated, deep well and normal Standard microplates. Additionally, options for microplate colors include clear , black, white or black with clear bottom and white with clear bottom for absorbance microplate reader applications. Find the best microplates for your lab in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.High-Throughput ScreeningHigh-throughput screening (HTS) is an automated drug discovery technique for identification of active compounds against a compound library. Use HTS readers and integrated assay preparation / analysis workstations to screen your compounds. Identify active compounds against various HTS libraries, including membranes, proteins and peptides and HTS cell lines. Find the best high-throughput screening products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.Colony CountingFluorescenceThe 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.Fluorescence MicroscopyFluorescence microscopy has become an essential tool in biology, as well as in materials science. The application of many fluorochromes has made it possible to identify cells and sub-microscopic cellular components with a high degree of specificity. Using multiple fluorescence labels, different probes can simultaneously identify several target molecules.ScreeningUsing robotics, data processing and control software, liquid handling devices and sensitive detectors, screening allows a researcher to quickly conduct millions of chemical, genetic or pharmacological tests.Cancer ResearchAlthough cancer is often referred to as a single condition, it actually consists of more than 100 different diseases. Microscopy, mass spectrometry, high throughput sequencing and flow cytometry are some of the most common techniques employed in cancer research labs.
High-throughput fluorescent colony formation assay