Utility of 405nm-Excitable Dyes in High Content Screening Using an Acumen Explorer Microplate Cytometer
Poster May 03, 2006
Sarah Payne, Tristan Cope, Christopher Lupton, Jeffrey T. Hung and Paul Wylie
The Acumen Explorer is a fluorescent microplate cytometer equipped with either a 405nm or 488 nm laser. The instrument offers rapid read and analysis times of individual cells within 96-3456 well plates (typically 5-10 minutes), making it compatible with the sustained use of high content methods in primary screens. In addition, small file sizes (>50kB in screening mode) are produced, therefore removing the requirement for expensive data storage solutions in compound screening programs.
In this study, we have used an Acumen Explorer equipped with a violet 405nm laser in conjunction with a selection of shorter-wavelength amine-reactive fluorophores (Invitrogen, Molecular Probes) to demonstrate a greater utility for blue fluorescent probes within high content assays.
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