MetroFlow 2021 Virtual
October 26th, 2021
ASCP CMLE Forms 1, 2, 3 (complete the forms and email a copy to MetroflowCMLE@gmail.com)
Recording Welcome & Introduction
Christina DeCoste, Director, Flow Cytometry Resource Facility at Princeton University & President, MetroFlow Steering Committee
Full Spectrum & Conventional Flow Cytometry
No Recording "Theoretical and practical aspects of autofluorescence in Cytometry: Impact on signal resolution, panel and assay design."
Alexis Perez Gonzalez, PhD, Manager, Melbourne Cytometry Platform at University of Melbourne
Recording "Fluorophore innovation for full spectrum flow cytometry"
Recording "Enabling Full Spectrum Panel Design: Novel BioLegend® Dyes and Experimental Considerations"
Recording Full Spectrum Panel Round Table Discussion
Recording Cytek Biosciences "Full Spectrum Profiling (FSP™) lets you see it, now Cytek® Aurora CS will let you sort it!"
Recording Nexcelom Bioscience "Applying ISO Cell Counting Standards to Improve Quality of Cell Counting Measurements"
Recording Immudex "Unravelling the specificity of T cells and beyond with Dextramer ® technology"
Coming Soon Abcam "FirePlex® multiplex assays – a biomarker discovery and screening technology platform"
Coming Soon "Semiconducting Polymer Dots (Pdots)"
Daniel Chiu, PhD, Professor of Chemistry and Bioengineering at The University of Washington
Recording "StarBright UltraViolet Dyes, increase your panel size with ease"
Recording "CAR Detection Reagents: Enhancing the Quality of Identifying CAR Surface Expression"
Coming Soon Reagents Panel Round Table Discussion
October 27th, 2021
ASCP CMLE Forms 1, 2, 3 (complete the forms and email a copy to MetroflowCMLE@gmail.com )
Recording "Adventures in Full Spectrum Flow Cytometry"
Christopher Hall, BSc(Hons), MSc, Senior Flow Cytometry Technical Specialist at the Flow Cytometry Facility at the Babraham Institute
Recording "Automated data analysis in flow cytometry: a “clinical-only” option?"
No Recording "Mastering cytometry data analysis with OMIQ"
Recording Data Analysis Panel Round Table Discussion
Recording De Novo Software "Moving from Data to Results with FCS Express"
Recording Fluidigm "CyTOF® XT: The evolution of high-parameter cytometry"
Recording Curiox Biosystems "Next-Generation Flow Cytometry Cell Preparation with Laminar Wash Technology"
Recording Agilent Technologies "Utilization of novel silicon photomultiplier detectors for improved performance in NovoCyte flow cytometers"
The Future of Cell Sorting
Recording "Cell sorting: a guide for the perplexed"
Rui Gardner, PhD, Head, Flow Cytometry Core Facility Lab Member, Immunology, Memorial Sloan Kettering Cancer Center
Recording "Microfluidic Gentle Cell Sorting for Intact Nuclei Sample Preparation"
Recording "Enrichment of microRNA Using a Novel Avalanche Photodiode-based Benchtop Cell Sorter"
Recording Cell Sorting Panel Round Table Discussion
Alexis Perez Gonzalez, PhD
Manager, Melbourne Cytometry Platform at University of Melbourne
The results presented and discussed in this lecture derive from a benchmark study comparing the sensitivity of Aurora spectral analysers against conventional cytometers at the University of Melbourne Cytometry Platform. Fluorescence sensitivity was assessed via Rainbow 8-peaks beads dim peaks resolution and stain index of human CD4+ cells stained with a broad collection of fluorochrome conjugates (26), differing in excitation, emission wavelengths, brightness and spectral signatures. The comparison of CD4-fluor brightness on fresh vs. fixed human blood revealed the negative effect of autofluorescence on signal dynamic range, resolution and thus the perceived brightness and ranking of fluorochromes. The operational brightness of fluorochromes in cytometry is not only dependent on properties of the dye (extinction coefficient and quantum efficiency) and the instrument (optical configuration and detectors sensitivity), but on the autofluorescence of the cells they label. The autofluorescence extraction feature gives Cytek Aurora a clear advantage in the resolution of fluorescence carried by highly autofluorescent cells, with the instrument showing the best performance across platforms in terms of human CD4 resolution on autofluorescence-rich regions. The benchmark was further extended into a biologically relevant setting, with the comparison of dim tdTomato resolution in Cytek Aurora 5L vs. Beckman Coulter CytoFLEX 6L, as part of a study on the tissue and cellular distribution of a barely known innate immunity marker. Our results reveal the critical role of autofluorescence extraction on the detection of dim fluorescence on highly autofluorescent cell subsets, and the clear advantage this feature gives Aurora instruments over highly sensitive platforms (i.e. CytoFLEX) that lack the autofluorescence extraction feature or the capacity to measure intrinsic cellular autofluorescence in fine details. The lecture will discuss the theoretical basis for the impact of Autofluorescence on signal resolution, describe several strategies for the isolation of unique autofluorescence signatures and illustrate methods for subset-specific autofluorescence extraction in both spectral and conventional analysers.
Daniel Chiu, PhD
Professor of Chemistry and Bioengineering at The University of Washington
Fluorescence based techniques, from cellular imaging to molecular assays, have become an indispensible tool kit in both basic cellular studies and in clinical applications and in vitro diagnostics. Organic dyes appear to be the most versatile fluorophores used so far in these applications. But the intrinsic limitations of conventional dyes, such as low absorptivity, limited multiplexing capability, and poor photostability, have posed difficulties for the further development of high-sensitivity techniques and high-throughput assays. As a result, there has been considerable interest in brighter and more stable fluorescent probes. We have developed and reported a new class of fluorescent probes based on semiconducting polymer nanoparticles called Pdots. This talk will highlight some of our recent advancements in developing new generations of Pdots for biomedical applications, with particular emphasis on flow cytometry.
Christopher Hall, BSc(Hons), MSc
Senior Flow Cytometry Technical Specialist at the Flow Cytometry Facility at the Babraham Institute
With the proliferation of full spectrum flow cytometers on the market (and in our facility at Babraham) I have been investigating how unmixing works and if there are any novel (or forgotten) analysis methodologies that could be applied to full spectrum flow cytometry data. In this talk I will discuss these with you.