We added a second flow cytometer. The Attune NxT has two lasers, 488 nm and 637 nm, and can detect seven fluorochromes, in addition to forward and side scatter. Examples of fluorochromes that can be detected simultaneously include FITC, PE, PE-Cy5, PE-Cy7, APC, AlexaFluor 700 and APC-Cy7. The instrument can accept samples from a traditional 12 x 75 mm tube as well as 96-well and 384-well plates. The instrument aspirates the samples volumetrically, allowing for accurate cell counting. The Attune has automated start-up and shut-down procedures, allowing experienced users 24/7 access to the instrument.
The BD LSR II underwent a major upgrade recently. The upgrades were made possible by support from the K-INBRE Core Facility Award (National Institute of General Medical Sciences (8P20GM103418)), the Office of the Vice Chancellor for Research, and the COBRE entitled, “Novel Approaches for the Control of Microbial Pathogens” (P30 GM103326).
The LSR II now has four lasers and is capable of detecting sixteen fluorochromes, upgraded from three lasers and nine fluorochromes. With the addition of the 552 nm laser, we are now capable of optimally detecting many of the newest commercially available fluorescent proteins, such as mCherry, mStrawberry, and Red Fluorescent Protein. The 552 nm laser will also permit optimal detection of PE and PE-conjugated fluorochromes. You will notice that these dyes will have a significantly brighter signal than in your previous experiments and you may need to re-titer antibodies conjugated to these fluorochromes.
We also expanded our ability to detect fluorochromes excited by the 405 nm laser, such as the newly developed Brilliant Violet™ (BD Biosciences) and eFluor® (eBioscience) series. We can now detect six fluorochromes excited by the 405 nm laser instead of two fluorochromes. A dye selection guide can be found at http://www.kumc.edu/flow/dye-selection-guide.html.
Our exciting news for this issue is that the COBRE grant that provides a substantial level of financial support was funded. This grant will be used to support the FCCL for the next five years as we continue to strive for sustainability. What does this mean for you? Without COBRE, we would be unable to support a user fee structure that is comparable to that of other institutions. To highlight the importance of COBRE and institutional support, we will redesign the invoices so that you will see how COBRE and KUMC support all users on campus. Because COBRE supports everyone, we kindly request that all users add the following statement to all manuscripts containing data obtained from FCCL instrumentation: “We acknowledge the Flow Cytometry Core Laboratory, which is sponsored, in part, by the NIH COBRE program of the NIGM P30 GM103326.”
During the next five years, we will seek to upgrade and add instrumentation that will support the KUMC scientific community. We always welcome suggestions for new flow cytometry-related instrumentation and upgrades. Further, we will gladly facilitate the preparation of instrumentation grants related to technologies appropriate for the FCCL.