Discrimination of spectrally overlapping fluorochromes using Sony Corporation’s (SP6800) Spectral Analyzer
Poster May 09, 2014
Divekar AA, Lee MJ, Kakuta M, Nitta N, Furuki M, Yang XF and Ransom JT
A maximum of seven colors can be separated using traditional 2-laser flow cytometers and it is required to have the necessary band pass filters to separate these fluorochromes. In addition, these systems cannot differentiate between spectrally overlapping fluorophores such as PerCP and PerCP/Cy5.5; or FITC, PE and Alexa Fluor 514® (A514). SP6800, a novel spectral analyzer from Sony Corporation, uses an innovative optics system that collects all emitted light and eliminates the need for band pass filters. We performed whole blood staining using a panel of 7-10 markers, which included significantly overlapping fluorochromes PerCP and PerCP/Cy5.5 and A514, PE and FITC and analyzed it on the SP6800. We were able to clearly identify CD56+ (PerCP/Cy5.5), CD19+ (PerCP) and CD3+ (A514) cells. Further comparison of a 7-color panel consisting of CD3 FITC, CD56 PE, CD19 PerCP, CD8 PE/Cy7, CD16 APC, CD4 Alexa Fluor 700® and CD62L APC/Cy7 on the BD LSRFortessa™ system and SP6800 gave similar results. Our data show that the SP6800 can perform all the analysis similar to a traditional cytometer with the added feature of separating spectrally overlapping fluorophores.
Quantitative Cell-Based Bioassays for Individual and Combination Immune Checkpoint Immunotherapy TargetsPoster
The human immune system is comprised of a complex network of immune checkpoint receptors that are promising new immunotherapy targets for the treatment of a variety of diseases including cancer and autoimmune-mediated disorders.READ MORE
Reporter Bioassays to Assess Therapeutic Antibodies for Immunotherapy ProgramsPoster
Immunotherapy, also called biologic therapy or biotherapy, stimulates certain parts of the immune system to fight diseases such as cancer. Important drug targets in immunotherapy include: Co-inhibitory receptors, such as PD-1/PD-L1, CTLA-4, LAG3, Tim3; and co-stimulatory receptors, such as GITR, CD40, OX40, 4-1BB.
Current approaches to assaying these targets are cumbersome and variable. Here we offer an improved in vitro bioassay approach.
Modelling CLL cell and T-cell Migration in a Dynamic Circulating Model of CLLPoster
We have recently developed a novel circulating model of chronic lymphocytic leukaemia (CLL) that mimics the transient interactions that take place between circulating lymphocytes and vascular endothelium. Here we show that both normal and malignant lymphocytes actively underwent transendothelial migration.READ MORE