2015 Program Commercial Tutorials

Saturday, 27 June 2015

Cell Sorting and Biosafety - Protecting You and Your Research Samples
BD Biosciences
2350 Qume Drive
San Jose, CA  95131
Phone: 877-232-8995
Email: answers@bd.com
Web: www.bdbiosciences.com
1245 – 1345 – Alsh Room
Presenter:  Morgan Blaylock and Jens Fleischer

Biological safety in flow cytometry is a growing issue for core laboratories concerned about the potential incidental exposure of operators to biological samples.   Scientific studies have demonstrated that there is an increased health risk from aerosol generation when performing cell sorting experiments on biohazardous material. Therefore a higher biosafety level of containment may be required during sorting than the one recommended for the infectious material alone.  Containment includes the use of a Biosafety Cabinet that can pass international biosafety standards for product, personnel, and environmental protection.  In this presentation we will show examples of product, personnel, and environmental safety testing for research use in accordance with the testing criteria defined in the national norms. In this context, research instrument solutions from BD Biosciences will be presented and usability for biosafety applications discussed.
ChipCytometry: A New 65-plex Immune Monitoring Panel, Exciting New Applications and Instruments
Bosestrasse 4, Saxonia
Leipzig  04109
Phone: 49 15152385628
Email: info@zellkraftwerk.com
Web: www.zellkraftwerk.com/cyto.html
1245 – 1345 – Boisdale Room
Presenter:  Christine Hennig
ChipCytometry is a high-content cytometry platform combining the unsurpassed quantitative phenotyping ability of flow cytometry with the unparalleled information depth of microscopy. Besides some features that are quite similar to conventional flow cytometry, three technology features make ChipCytometry an exciting technology for explorative high-content analysis:
1. Long-term sample storage/ no sample consumption: ChipCytometry uses microfluidic chips enabling biomarker-preserving long-term storage of samples for a period of at least 20 months. Cell storage is particularly useful for precious samples like patient samples, rare cells and (multicenter) clinical trials because cells are not destroyed during transport, storage and analysis and can be stored for further tests exploring more and more markers on these pre-analyzed samples.
2. High-content cytometry: ChipCytometry enables highly multiplexed biomarker assays. Zellkraftwerk presents a 65-plex immune monitoring assay for comprehensive human immune cell phenotyping, and a 39-plex murine immune monitoring panel.
3. Tissue Cytometry: ChipCytometry can work with as sorts of cell specimens as well as with a broad range of tissue types.
At CYTO 2015 Zellkraftwerk will launch the Cytobot™, the first fully automated high-throughput ChipCytometer including fully automated data processing and lab management software. The Cytobot™ is a new instrument designed for core facilities, whereas the ZellScanner ONE (launched in 2014) is a compact benchtop instrument for low to medium sample throughput.
Join us for an introduction to the future of “precious samples cytometry”.
Typical German lunch bags will be served on a first come first serve basis.

Maximize your FlowJo:  Applying FlowJo Enterprise to a Clinical Research Study
FlowJo, LLC
385 Williamson Way
Ashland, OR  97520
Phone: 541-201-0022
Email: caitlinf@flowjo.com
Web: www.flowjo.com

1245 – 1345 – Carron Room

Presenter:  Jack Panopoulos, Ph.D. and John Quinn, Ph. D.

Clinical research studies present unique software challenges to data management, analysis, reports and sharing.  Advances in technology have allowed them to grow in complexity and have produced richer data sets to mine.  At the same time the rigor required for work related to clinical research has not depreciated.  FlowJo Enterprise enhances the clinical research environment by providing centralized and consolidated analysis pipelines, facilitating push-button report generation.  This centralized server based system can assist with the compliance aspects of clinical research by generating electronic logs of all data and associated derivatives’ activity from acquisition to analysis and reporting. Server logs can easily be queried by an authorized administrator for various activities, including data upload, workspace manipulation, security access and reports. Restricted access to data, reports and derivatives is regulated by a high- level administrator, further increasing security and compliance with good data management practices.
FlowJo’s workshop tutorial will include a review of federal compliance guidelines and provide an overview of FlowJo Enterprise’s utility as applied to a common clinical research study.

ACEA NovoCyte™ System - Redefining Benchtop Flow Cytometers with High Performance, Efficient Tools in Customizable Budget Friendly Platforms
ACEA Bioscience
6779 Mesa Ridge Road, # 100
San Diego, CA  92121
Phone: 858-724-0928
Email: info@aceabio.com
Web: www.aceabio.com
1245 – 1345 – Seminar Suite
Presenters:  Derek Davies, Cancer Research UK; Alan Saluk, The Scripps Research Institute; Andy Filby, University of Newcastle
In this workshop key opinion leaders will present complex multi-parameters studies, comparative studies as well as microparticle applications using the new innovative NovoCyte flow cytometer from ACEA.

Sunday, 28 June 2015

Optimizing Strategies for Reliable Multicolor Flow Data
BD Biosciences
2350 Qume Drive
San Jose, CA  94087
Phone: 877-232-8995
Email: answers@bd.com
Web: www.bdbiosciences.com
1245 – 1345 – Alsh Room
Presenter:  Robert Balderas
Flow cytometry has been a powerful tool for the immunology laboratory and continues to advance from using 3-4 parameters to using >15 parameters and beyond in a single panel. The innovations have made multi-color flow cytometry more accessible to researchers worldwide working on other areas of biology such as marine sciences, microbiology and stem cell biology.  In this tutorial we will review various approaches that can be taken to ensure data quality and reproducibility whether we are using 4 or 15 parameters. We will explore how choices in instrument capability, reagent selection and approach to setup strategies can improve our confidence in data and simplify workflows for both cell analysis and cell sorting. We will also look at experimental approaches for studying rare/dim populations, and facilitating transitions from a low to high complexity panel. These multicolor optimization strategies for cell analysis and sorting populations of interest make it easier for researchers to generate good quality data and drive discovery.
Multicolor Flow Cytometry on your Benchtop: Now More Sensitive and Versatile with the Guava easyCyte™ Systems
Merck Millipore
Boulevard Industrial Park, Padge Road
Nottingham, NG9 2JR
United Kingdom
Phone: 44 115 943 0840
Web: www.merckmillipore.com
1245 – 1345 – Boisdale Room
Presenters:  Katherine Gillis and Jessica Reed
The guava easyCyte™ microcapillary flow cytometry systems are simpler to operate than traditional sheath-fluid based instruments and are far easier to maintain. They utilize small sample volumes, generate minimal waste, and have lower operating costs. In this workshop, attendees will learn how the simplicity and sensitivity of the improved guava easyCyte™ systems enhances data collection and analysis. Please come visit us and learn more!

(1)  Microfluidic Droplet-free Sorting: the Gentle Way to Purify Cells
Miltenyi Biotec GmbH
Friedrich-Ebert-StraBe 68
Bergisch Gladbach 51429 
Phone: 49 2204 8306 0
Email: macs@miltenyibiotec.de
Web: www.miltenyibiotec.de

1245 – 1345 – Carron Room
Presenter: John Foster and Christian Peth
Microfluidic sorting offers a number of advantages compared to conventional state of the art jet-in-air cell sorters. High sample throughput, purity, yield and low cell stress can be achieved simultaneously. The benchtop cell sorting instrument MACSQuant® TytoTM uses a sterile disposable cartridge with a microfluidic chip to purify samples in a closed environment. The heart of the microchip is the world’s fastest valve that enables fast and precise flow switching in less than 10µs. Therefore, sort rates up to 50.000 events per second can be achieved. The droplet-fee microfluidic approach minimizes cell stress during the sort process resulting in high cell viabilities and functionality. In addition, the absence of aerosols in combination with a closed cartridge ensures operator safety during operation. Simple workflows without complicated setup and alignment procedures allow for sorting with just a few mouse clicks.
(2)  Not Your Average Flow: Creating Customized Flow Cytometry Solutions Using the MACSQuant Platform
Presenter: Jeffrey Carrell
As flow cytometry core lab managers, we are challenged with providing access to methods and technology across global sites, often without full time on-site core staff. To this end, we advocate for standardized instruments and methods enabling the sharing of protocols across a large organization. Our solution was to evaluate an easy-to-operate and highly capable cytometer platform that was well-supported with a line of reagents and assay kits. We chose the Miltenyi Biotec MACSQuant platform as it met all of these criteria. We describe several projects we are developing in collaboration with Miltenyi, such as: creating a custom barcoded antibody cocktail, incorporating an automated data collection and analysis template for immunophenotyping; building a multi-assay toolkit for the evaluation of biotherapeutics; and producing a series of training videos that we will deploy across global sites. These customized solutions have led to faster data generation and real-time analysis, improved access to flow cytometry methods for non-experts, and more efficient use of core staff time. Most importantly, the platform allows for global harmonization of capabilities, enabling more of our users to conduct sophisticated flow cytometry experiments.
Advances in High Throughput Flow (HTF): The Ideal Platform to Enhance Productivity in Immunology Research
IntelliCyt Corporation
9620 San Mateo Blvd., NE
Alburquerque, NM  87113
Phone: 505-345-9075
Fax: 866-781-3140
Email: support@intellicyt.com
Web: www.intelliCyt.com
1245 – 1345 – Seminar Suite
Presenter: Joseph Zock

Key features of the iQue Screener make it uniquely capable to screen both cells and beads, separately and together. By sampling as little as 1 uL from 96, 384 and 1536 well plates, it allows for dramatic reduction in assay volumes to as little as 5 μL. Combined with its ability to analyze an entire 384-well plate in less than 20 minutes, multiplexed screening with precious samples is a reality. ForeCyt Software manages the acquisition and analysis of highly multiplexed data and provides insightful data visualizations at per cell, per well and per plate levels. These features and more place the iQue Screener as the ideal platform for combinatorial immunotherapy characterizations and immunotarget screening. In this tutorial you will see how the Integrated Screening Solution that iQue keystones makes screening of Immunology and Immuno-oncology applications a reality. You will also learn about recent advances to the iQue Screener platform.

CytoFLEX – How New Technology Translates into Exciting Applications
Beckman Coulter
22, rue Juste-Olivier
Nyon  1260
Phone: 49 2151 333-723
Web: www.beckmancoulter.com
1245 – 1345 – Lomond Auditorium
Presenters: Andreas Spittler, Medical Univeristy of Vienna and Yong Q. Chen, Chief Technical Officer, Beckman Coulter Life Sciences

Beckman Coulter has recently launched a new Cytometry platform called CytoFLEX. Its revolutionary technology leads to unmatched performance and results. Learn how CytoFLEX can bring your research to the next level.

Monday, 29 June 2015

Overcoming Challenges in Cellular Analysis: Rare Event Detection in the Innate and Adaptive Immune System
Thermo Fisher Scientific
3 Fountain Drive, Inchinnan Business Park
Paisley Renfrewshire, PA4 9RF
United Kingdom
Phone: 44 1418145955
Web: www.lifetechnologies.com
1245 – 1345 – Alsh Room
Presenters: Ryan P. Larson, PhD; Andrea Cossarizza, MD/PhD, University of Modena and Reggio Emilia School of Medicine, Italy
Analysis of rare cell types in complex biological systems is a major challenge in biomedical research. The multi-parameter capability and high analysis rate make flow cytometry one of the key technologies in identifying and characterizing rare cell types. This seminar will discuss the application of this technology to the analysis of rare cell types in the immune system. Acoustic focusing cytometry allows for high sample collection rates without compromising data integrity, thus allowing for the rapid analysis of rare events in dilute samples or in a no-lyse, no-wash whole blood format.  We have developed multiple techniques for no-lyse, no-wash assays for whole blood analysis with minimal sample processing along with assays to characterize phagocyte phenotype and function utilizing pHrodo® BioParticles® phagocytosis/phagosome acidification assay.  In addition, we will discuss the identification and functional characterization of innate lymphoid cells, circulating antigen-specific lymphocytes, and innate-like cells (such as invariant natural killer T cells and circulating endothelial cells).

Morphological Analysis, Internalization, Colocalization:  What Does Imaging Flow Cytometry Reveal?
Merck Millipore
Boulevard Industrial Park, Padge Road
Nottingham, NG9 2JR
United Kingdom
Phone: 44 115 943 0840
Web:  www.merckmillipore.com
1245 – 1345 – Boisdale Room
Presenter: Sherree Friend, PhD, Amnis-part of Merck Millipore
Imaging flow cytometry combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy. In this workshop you will learn how the features and benefits of the Amnis® ImageStream®X Mark II and FlowSight® imaging flow cytometers can be applied to your research. Please come visit us and learn more!
Analysis of CD163 mRNA for Monocyte Pathobiology in Systemic Juvenile Idiopathic Arthritis; and Integration of PrimeFlow RNA Assay into a Flow Cytometry Core
eBioscience, an Affymetrix Company
Campus Vienna Biocenter 2
Vienna 1030
Phone:  43 1 796 4040 305
Fax: 43 1 796 4040 400
Email: info@ebioscience.com
Web:  www.ebioscience.com

1245– 1345 – Carron Room
Presenters:  Sherry L. Thornton, PhD and Monica DeLay
Integration of PrimeflowTM technology into a shared facility will be discussed. Examples of using this technology will focus on examination of macrophage differentiation of monocytes in Systemic Juvenile Idiopathic Arthritis, SJIA. SJIA, is a chronic inflammatory disease of childhood with prominent extra-articular features, including prolonged spiking fevers, rash, lymphadenopathy and serositis.  Macrophage activation syndrome occurs in approximately 10% of SJIA patients, and is a potentially fatal episode of overwhelming inflammation with emergence of hemophagocytic macrophages. SJIA monocytes exhibit both classically activated M1 attributes as well as markers associated with alternatively activated M2 macrophages, such as CD163. PrimeflowTM technology was used to detect CD163 mRNA in THP-1 monocytes, as protein levels assessed by CD163PE antibody binding show minimal cell surface expression.  Upon stimulation of THP-1 cells with IL-10, CD163 mRNA was increased approximately 3-fold. Regulation of CD163 via microRNAs is also under investigation and will be discussed.
Comparison of QbSure and CS&T QC
Cytek Development
4059 Clipper Court
Fremont, CA  94538
Phone:  510-657-0102
Email: cytekdev@cytekdev.com
Web:  www.cytekdev.com
1245 – 1345 – Seminar Suite
Presenter: Eric Chase PhD
QbSure and CS&T QC programs use different measurements of instrument performance to determine pass/fail results. QbSure uses the resolution limit (number of MEFLs required for full resolution from noise) and the hydrodynamic CV of a bright bead for its pass/fail criteria.
CS&T uses a shift in the target PMT voltage and the hydrodynamic CV of a bright bead for its pass/fail criteria. The shift in the target PMT voltage is with respect to a baseline measurement, so is a relative measurement. The resolution limit measurement is an absolute criterion. Both programs track instrument performance over time. However, the QbSure program’s use of a resolution limit to evaluate each parameter allows a direct between-instrument comparison of performance. Without absolute criteria, CS&T cannot provide feedback when instrument performance is poor relative to other instruments.
Extensions of QbSure software to DiVa data files will be discussed. This will allow the measurement of the resolution of instruments running DiVa.
Novel Approaches to Extra Cellular Vesicle Detection and Sorting
Beckman Coulter.
22, rue Juste-Olivier
Nyon  1260
Phone:  49 2151 333-723
Web:  www.beckmancoulter.com
1245 – 1345 – Lomond Auditorium
Presenter: Laura Pajak, Beckman Coulter Life Sciences
Andreas Spittler, M.D. (Medical University of Vienna), Jennifer Jones, Ph.D. (NIH) and Carley D. Ross, Ph.D. (Beckman Coulter) will discuss how to utilize the Astrios EQ for extracellular vesicle sorting. Instrument configuration and results obtained will be shown when sorting extracellular vesicles from cells such as HeLa and whole blood.

Tuesday, 30 June 2015

Calculation of Sort Recovery and Using Rmax as an Improved Measure of Sorter Performance
Bio-Rad Laboratories
2000 Alfred Nobel Drive
Hercules, CA  94547
Phone:  510-741-4494
Web:  www.bio-rad.com
1245 – 1345 – Alsh Room
Presenters: Dr. Rui Gardner and Carol Oxford
At Bio-Rad, we are committed to measuring the most important parameters to determine optimal sorter performance. A new method of sorter performance has been described to evaluate sorter performance based on the measurement of sort recovery. The method precludes the need for absolute counting and re-sampling the sorted fraction, and requires only determining the ratio of target and non-target populations present in the original pre-sort sample and the non-sorted fraction in the waste stream. The method involves the calculation of Rmax, which is a measure of the maximum recovery expected in the conditions the sort was carried out, i.e., for a given instrument configuration and sort setup. Rmax can also be used to troubleshoot the instrument, to control daily drop-delay determination, and to evaluate and troubleshoot sample preparations by comparing them with an ideal sample. Rmax is a measure of the instrument’s maximum recovery, is dependent on instrument performance, and Independent of sample-related losses. It is calculated based on the cytometric analysis of the original sample, and avoids sampling and absolute counting of the sorted product. Bio-Rad Laboratories patented ProDrop process provides automated calculation of the drop delay based on actual events In the waste stream, providing a recovery based drop delay calculation.
Spectral Cytometry Session: Introducing the Sony SA3800 High-throughput Spectral Analyzer and its Unmixing Algorithms
Sony Biotechnology Inc.
1730 North First Street
San Jose, CA  95112
Phone:  800-275-5963
Fax: 408-352-4130
Email:  sales@sonybiotchnology.com
Web:  www.sonybiotechnology.com
1245 – 1345 – Boisdale Room
Presenter:  Marsha L. Griffin Ph.D., MPH

In this talk we will focus on the features of the newest addition to the family of spectral analyzers and the details of Sony’s own spectral  unmixing algorithms.  Spectral flow cytometry is proving to be an exciting technology for cytomics and systems biology. Spectral flow cytometry differs from conventional flow cytometry in that the measured parameters for events are fluorescence spectra taken across all detectors as opposed to being primarily the fluorescence signal measured from one detector. This gives spectral flow cytometry capabilities and flexibility that far exceeds those of conventional flow cytometry.  Additional flexibility has been engineered into Sony’s latest analyzer, the SA3800, by extending to users software-driven high-throughput functions that adapt to both plate and well formats permitting walk-away acquisition for the busy scientist. 
The core functions of a spectral detection scheme facilitate the simultaneous independent measurement of signals and the data processing power to unmix such signals for real time visualization.  These fundamental differences enable spectral flow cytometers to perform applications that are not readily possible on conventional flow cytometers. Because a spectral flow cytometer measures the whole fluorescence spectrum for each fluorophore, overlapping fluorophores can be resolved based on spectral shape allowing for set-up of panel combinations and the use of markers that would not be resolvable by conventional flow cytometry. Additional options for analysis include unique spectral graphs for each laser line.  These graphs enable the visualization of different cell subsets and data checking.

Frontiers in Mass Cytometry: Roadmap for Discovery of Immune Signatures that Correlate with Clinical Recovery after Surgical Trauma
Fluidigm Corporation
7000 Shoreline Court
S San Francisco, CA  94080
Phone: 408-900-7224
Web: www.fluidigm.com
1245 - 1345 – Carron Room
Presenters:  Brice Gaudilliere, MD/PhD, Stanford and Tad George, PhD, Fluidigm

Infection, cancer, tissue injury and other perturbations to human health provoke profound immune responses that influence clinical outcome and recovery.  Surgery represents one such perturbation that is performed on over 100 million patients annually in Europe and the United States, and discovery of immune signatures that correlate with recovery could provide a foundation for targeted therapies that improve postoperative recovery. 
Mass Cytometry’s ability to simultaneously measure 40+ phenotypic and functional proteins at the single cell level for millions of cells per experiment uniquely enables system-wide analysis of the immune system and therefore was used to search for immune signatures that correlate with clinical recovery from surgery.  Using 23 surface markers and 11 intracellular signaling markers, the study quantified endogenous immune signaling responses of patients undergoing surgery. Single-cell barcoding of patient samples and normalization of the mass cytometry data was essential to ensure that observed sample variability in endogenous signaling responses was in fact due to patient-specific factors rather than experimental conditions.   Using this study as a substrate, the tutorial will discuss practical aspects of palladium-based sample barcoding and data deconvolution when implementing mass-tag barcoding in a clinical study.
In this tutorial we will also describe recent advances in instrumentation and reagent development that continue to expand systems-level discovery at the single cell level through mass cytometry.
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