High Throughput High Content Screening using a Multiwavelength Microplate Cytometer
Poster Feb 01, 2007
Sarah Payne, Paul Wylie and Wayne Bowen
Fluorescence microplate cytometersare designed for a screening environment as they are capable of analysing up to 300,000 high content data points in 24 hours (1536 well format). They deliver the object recognition capabilities of a microscope-based CCD imager with the fast data collection rates and small data file sizes associated with bulk fluorescence readers. Statistical quality of screening data is enhanced by whole well scanning capability; whilst assays requiring larger fields of view than those offered by microscope objectives may be transferred into a higher throughput, automated format.
The Acumen eX3, a multiwavelength microplate cytometer, offers 405, 488 and 633 nm laser excitations in one instrument. This advancement significantly extends the range of fluorescent reagents that can be combined in multicolour, multiplexed assays over a wavelength range for excitation that is similar to that of white light source instrumentation. Thus, it permits seamless transfer of high content assay protocols, developed using CCD-based imaging devices, onto the Acumen eX3 for screening.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE
During early drug discovery, the study of metabolism plays an essential role in determining which drug candidates move forward into development and later stages. As an alternative to traditional Data Dependent Acquisition (DDA), the use of MSE/All Ions Fragmentation (AIF) has become common in metabolite identification workflows for the analysis of metabolic hot spots. Here we present a solution for analysis of MSE/AlF in metID studies.READ MORE
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