William Godfrey, Ph.D., Manager, Reagent and
Application Development, Beckman Coulter, Inc The
three Beckman
Workshop Description: In recent years flow cytometers have become more powerful with the addition of large laser palates and multiple flourochrome options. This talk will look at the considerations a researcher needs to make while designing high color applications. We will look at the importance of titration, spectral overlap, dye selection, tandem dyes, background control and sample prep when constructing a complex cocktail or panel. Specific panel and cocktail examples to be discussed include L/L, Cell Signaling and NK. We will also examine how any new dyes will effect panel development.
Presenter:
Chris Novak, Global Product Manager, Flow Cytometry, Beckman Coulter, Inc.
Workshop Description: The Beckman Coulter MoFlo™ Astrios™ sorter is a high-end 7-laser system which allows for the ultimate in cell sorting flexibility. In this tutorial we will illustrate the system´s features looking at bio containment, automation, flexibility and operator ease-of-use. Cells samples analyzed will include Circulating Hematopoietic Progenitor Cells and a comprehensive sort of T Cell subsets. The data we review demonstrates the performance of the system with respect to, six-way sorting, sensitivity, sort speed, purity and recovery of sorted populations.
Workshop Description:
Through innovations in optical and electronics design, cytometers have progressively improved their ability to reliably resolve very small particles and dimly fluorescent signals. The Beckman Coulter Gallios system employs short optical paths and high power lasers, delivered directly through air to increase the available excitation energy. Additionally, highly efficient collection optics are used to ensure that the emitted light loss is minimized. Complementing these innovative optical designs is the highly resolved electronics that use a 20 bit integral signal and 40MHz sampling rate. The Gallios is therefore able to maintain very high sensitivity without compromising the speed of data acquisition. Other innovations, such as the collection of two angles of forward scatter, allows for much greater resolution between sub-micron particles.
The benefit of the application of these technologies is clearly evidenced by the superior resolution in dimly fluorescent bead populations that has been reproducibly demonstrated in comparison with a variety of other multi-color cytometers. Of course beads are just beads and unless these benefits are manifest in cellular analysis, they are of limited value to the cytometrist. Fortunately, a variety of enhanced cellular applications have been described that have been enabled or significantly improved using the Gallios system. Examples include the improved size resolution of both red cell microvesicles and platelet microparticles. In the context of fluorescence detection, applications that include vector transfection confirmation and complex cell signaling assays, were difficult using other systems have been enabled by the fluorescence resolution of Gallios. Finally, the speed of the Gallios expressed in events per second has been studied in comparison with other systems. It is clear, that while some systems suffer major reductions in the yield of detectable events, the Gallios is able to markedly reduce cellular data losses, even as the maximum event rate of 25000 EPS is reached.
These features of more highly resolved size and fluorescence signals, as well as high speed and yield in a 10 color flow cytometer, have resulted in tangible benefits in the performance of cytometric applications and in some instances, wholly new applications.
Beckman Coulter 2010 CYTO Workshops Presentations
Presenters
Presentation
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William Godfrey, Ph.D.,
Panel Design: Getting More "Multi" Into Your Multi-Color
Chris Novak
High Speed, High Color Six-way Sorting of Circulating Hematopoietic Progenitor Cells and Lymphocyte Subsets
Sharlene Wright
New Frontiers in High Sensitivity Cytometry
2010 CYTO Posters
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Posters
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1
Flow Cytometric Analysis of Multiple Signaling Pathways in LPS Stimulated Human Peripheral Blood Monocytes
T. Vincent Shankey
Advanced Technology Group, Beckman
Coulter, Inc., Miami, FL, USA Sue Chow
Division of Applied Molecular
Oncology, Princess Margaret
Hospital, Univ of Toronto, Toronto,
Ont., Canada David Hedley
Division of Applied Molecular
Oncology, Princess Margaret
Hospital, Univ of Toronto, Toronto,
Ont., Canada
2
Krome Orange
Felix Montero,
RavinderGupta, HashemAkhavan-Tafti,
Robert Eickholt, Mark Sandison,
Rhonda Federspiel, Laura Nieto
Gligorovsky, Franck Gaille, Jeffrey
Cobb1and Emmanuel Gautherot Beckman Coulter, Inc.,
Miami, FL, USA; Detroit, MI, USA;
Marseille, FRANCE
3
An Automated Cell Preparation Method For FlowCARE™ Pan-Leukocyte Gating (PLG)
Jin Zhang, ValentinQuesada,
Juan De Castro, Jennifer Zawislak,
Jorge Acevedo, Ed Jachimowicz, and
LilianaTejidor Beckman Coulter, Inc.,
Cellular Analysis Business Group,
Miami, FL