CYTO U Upcoming Webinars
Collapse SRL Best Practices:  Managing Data in a Flow Cytometry Core

This live webinar is the first in a series of Best Practices webinars.  It will give an overview of the important aspects in managing data in a Shared Resource Lab and provide examples of common practices used among some of the best SRLs from around the world,

Formats Available: Streaming, Live Webinar, Webinar + Archive
Original Seminar Date: March 16, 2017
On-Demand Release Date: Available Now

Approved Credit:
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information SRL Best Practices:  Managing Data in a Flow Cytometry Core
    Collapse SRL Best Practices Series:  Standard Operating Procedures (SOPs)
    The second of seven webinars in the SRL Best Practices Series
    Formats Available: Streaming, Live Webcast
    Original Seminar Date: March 28, 2017
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information SRL Best Practices Series:  Standard Operating Procedures (SOPs)
    Collapse Recycle
    Original Seminar Date: May 09, 2017

    Approved Credit:
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information Recycle
    Collapse Live Cell and Super-Resolution Optical Microscopy
    Formats Available: Recording, Live Webcast, Live Webinar & Streaming
    Original Seminar Date: May 09, 2017
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information Live Cell and Super-Resolution Optical Microscopy
    CYTO U Archived Webinars
    Collapse Using CellProfiler for Biological Image Analysis

    Mark-Anthony Bray, Ph.D. will present a webinar Using CellProfiler for Biological Image Analysis. Mark earned his B.S. in biomedical engineering from Tulane University in 1996 and his Ph.D. in biomedical engineering from Vanderbilt University in 2003, followed by postdoctoral work in cellular bioengineering at Harvard University. In 2008, he joined the Imaging Platform at the Broad Institute as a computational biologist under the direction of Anne Carpenter. Using his biology and image assay development expertise, he is now involved in the adaptation of image analysis software to specific image-based screens.

    Image-based screens increasingly require the use of automated microscopes which generates hundreds, if not thousands of images at a time. With such large image sets, automatic image analysis is more objective and quantitative and less tedious then visual inspection alone. This webinar is meant to instruct biologists in the use of CellProfiler, an open-source, freely-downloadable software package designed for analyzing such images via a user-friendly interface. Material will begin an introduction of CellProfiler, followed by details for the construction and use of a pipeline for analyzing typical image data. We will also include some of the more important considerations in cellular image analysis, by demonstrating the software on sample images as a realistic use case. Lastly, we will cover measurement export and preparation for additional analysis using  machine-learning tools.

    At the completion of this tutorial, the attendee will have knowledge of the following:
    • Understand the basics of the CellProfiler environment;
    • Design a simple pipeline using the graphic user interface;
    • Test their pipeline and adjust the settings for optimal performance.
     
    Who Should Attend:This tutorial is intended for beginners to quantitative cellular image analysis. No programming skills are required. However, basic knowledge of fluorescence microscopy and digital image acquisition is expected.
    Formats Available: Streaming
    Original Seminar Date: January 14, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Using CellProfiler for Biological Image Analysis
    Collapse Integrating Flow Cytometry and Single-Cell Transcriptomics

    Mario Roederer, Ph.D. will be presenting the webinar Integrating Flow Cytometry and Single-Cell Transcriptomics. Mario serves as Senior Investigator on the ImmunoTechnology Section, Chief on the Flow Cytometry Core, and Chief on the Nonhuman Primate Immunogenicity Core. The immune system is comprised of incredibly diverse sets of cells, each programmed to carry out overlapping sets of effector functions. Quantifying any one function provides an incomplete view of the immune response, as information about what other responses are generated is absent. Quantifying multiple responses is far superior, but when carried out on a bulk level, loses information about cellular heterogeneity, gene programs, and a myriad of interactions that may occur at the single-cell level. Since individual cells are the atomic unit of immune function, the maximum information content is achievable only by measuring these functions independently and simultaneously on a cell-by-cell basis. For this reason, flow cytometryis a powerful technology to assess immune function in settings like vaccination and pathogenesis. Nonetheless, current flow cytometrytechnology is limited to measuring the expression of ~16 proteins per cell. To extend the multiplexing of gene expression measurements, we now combine single cell sorting (based on cell surface phenotype) with highly-multiplexed qPCR for 96 or more genes. We choose to quantify lymphocyte-centric genes, including those encoding transcription factors, signaling molecules, effector;molecules, and regulatory molecules. On a single-cell basis, we can correlate protein expression with gene expression. Discordant results for the same gene reveal post-transcriptional regulatory mechanisms. We have identified gene signatures associated with vaccine-elicited T cells as well as with productively SIV-infected cellsin vivo. This technology gives us an unprecedented view into the complexity and range of immunological functions expressed by vaccine or virus-specific immune cells. Using this approach, we can search for correlates of clinical outcome based on either: quantitative gene expression; and/or cell subset representation, enumerated by groups of cells sharing gene expression profiles. These analysesgive us new insights into functional immune states in pathogenesis, treatment, and vaccination.

    Formats Available: Streaming
    Original Seminar Date: February 19, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Integrating Flow Cytometry and Single-Cell Transcriptomics
    Collapse The Signaling Networks Regulating Morphological Heterogeneity

    Chris BakalChris Bakal, Ph.D. will be presenting the webinar "The Signaling Networks Regulating Morphological Heterogeneity." Chris is a Dynamical Cell Systems Team Leader in the Division of Cancer Biology at the Institute of Cancer Research in London UK.  

    The Dynamical Cell Systems Team is using high-throughput functional genomic approaches in tandem with integrative computational technologies to understand the architecture and dynamics of signal transduction networks. He studies the biological switches that cause cells to change shape, become cancerous and spread around the body. By understanding how these switches work, scientists may one day find a way to control them through drugs or other therapies. Cells are able to assume a wide variety of complex shapes in order to carry out different roles and Dr Bakal's team examines the genetic and biochemical mechanisms that underpin these shape changes. Their research covers how both environmental and genetic variation affects cell shape. Ultimately, Dr Bakal aims to understand how normal and cancerous cells can adopt different shapes and why metastatic cancer develops in some people but not others.

    Formats Available: Streaming
    Original Seminar Date: March 20, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information The Signaling Networks Regulating Morphological Heterogeneity
    Collapse Enhancing the Shared Resource Laboratory through the Use of Social Media

    Ryan Duggan

    Ryan Duggan will be presenting the webinar "Enhancing the Shared Resource Laboratory through the Use of Social Media." Ryan serves as the Technical Director of the Flow Cytometry Core Facility at the University of Chicago (UCFlow).

    The past decade has seen a dramatic emergence of new media forms (such as social media) and web-(or"cloud") based tools that have had a significant impact on the ways in which individuals and organisations interact and conduct business. Like other areas of science and industry, shared resource laboratories (SRLs) have much to gain from leveraging these technologies. This webinar will share some of my experiences in utilising these tools. New media will be discussed in the context of enhancing SRL visibility and status and interacting with colleagues and customers. These tools will give your facility renewed confidence in providing high quality services in an efficient manner while expanding your user base within and beyond your own Institution.

    Formats Available: Streaming
    Original Seminar Date: April 16, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Enhancing the Shared Resource Laboratory through the Use of Social Media
    Collapse Beyond Polychromatic: Spectral Flow Cytometry

    John P. Nolan, Ph.D. will be presenting the webinar "Beyond Polychromatic: Spectral Flow Cytometry."  John is the Past-President of ISAC and he is currently a Professor at the La Jolla Bioengineering Institute/The Scintillon Institute in San Diego, CA.

    Since the earliest days of flow cytometry, researchers have sought to bring the spectral resolution of a fluorescence spectrometer to the analysis of individual cells in flow, but faced many limitations in optics, detectors, and electronics that made this impractical. Today, these limitations have been largely overcome and it is possible to collect the full fluorescence emission spectra of individual cells in flow cytometers that use high efficiency prisms of gratings to disperse the light collected from single cells across a detector array. The ability to collect the full fluorescence spectrum allows compensation to be replaced by spectral unmixing, which can provide more accurate estimates of label intensity and resolve fluorochromes with significant spectral overlap.  Moreover, spectral flow cytometry opens the door to a new era of flow cytometry instruments and applications. Spectral flow cytometers have simpler optical paths and fewer components than conventional flow cytometers, potentially resulting in lower cost and maintenance. More importantly, spectral flow cytometry enables the use of new types of labeling probes. For example, nanoparticles exhibiting surface enhanced Raman scattering (SERS) have attracted interest for their multiplexing potential. In this webinar, I will describe the essential elements of spectral flow cytometry and show examples of fluorescence and SERS detection.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: June 19, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Beyond Polychromatic: Spectral Flow Cytometry
    Collapse μManager: Open source software for microscope image acquisition

    Nico Stuurman, Ph.D. will be presenting the webinar "μManager: Open source software for microscope image acquisition" on Wednesday, July 23 at 12:00 pm ET. Nico is a Research Associate in the laboratory of Ron Vale at the University of California San Francisco where he is involved in microscopy and computer programming.

    Research microscopes are usually equipped with digital cameras and motorized equipment such as shutters, stages, and filter wheels, necessitating computer control. Several commercial solutions exist, which are often tied to specific hardware, are often available only on Windows, are difficult to extend and incorporate in novel workflows, and do not allow for inspection of the source code. We therefore developed μManager, a freely available open source software package for control of motorized microscopes, which runs as a plugin to ImageJ. μManager is cross-platform (Windows, Mac, and Linux), hardware agnostic (it works with an ever growing number of microscopes and peripherals and device support can be written by anyone), extensible (through scripting and plugin interfaces, as well as hooks to other environments such as Python and Matlab), and is easy to use.

    In this webinar I will give some background about the purpose and capabilities of μManager, demonstrate the most common ways μManager is used to acquire microscope data, highlight a few of the μManager plugins that extend its functionality, and show what is involved in configuring a microscope system.

    This webinar will be useful for microscopists considering the use of μManager as well as for μManager users who want to learn more about the capabilities of the software.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: July 23, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information μManager: Open source software for microscope image acquisition
    Collapse Forensic Flow Cytometry

    Jennifer Wilshire will be presenting the webinar "Forensic Flow Cytometry." Jennifer is the Assistant Manager of the Flow Cytometry Core Facility at Memorial Sloan-Kettering Cancer Center in New York, NY.

    Do you sometimes look at flow cytometry results and wonder if something went wrong, but can't put your finger on it? In this webinar we will take a "forensic" approach to study the most common issues that give bad flow cytometry data.  We will use case studies and real examples to illustrate the causes of bad data and how you can prevent it. Many topics will be covered including staining issues, compensation, controls, and sorting. The focus of this webinar will be on understanding the potential pitfalls in flow cytometry experiments and learning practical methods to prevent these issues. After participating in this webinar, attendees will have practical suggestions of ways to ensure flow cytometry experiments will yield good data and purely sorted cells.

    Formats Available: Streaming
    Original Seminar Date: August 05, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Forensic Flow Cytometry
    Collapse Standardization of flow cytometry assays: What for? How? Where does it lead us?

    Tomáš Kalina, MD. PhD, is currently Associated Professor at Department of Pediatric Hematology and Oncology, Charles University in Prague, 2nd Faculty of Medicine, Czech Republic.

    He is a founding member of EuroFlow consortium where he is responsible for coordination of the technical aspects and design of flow cytometry procedures. At present, he is actively involved in EuroFlow Primary Immunodeficiency workpackage and EuroFlow proficiency testing. Tomáš is also an ISAC Scholar.

    Webinar will focus on standardization of the flow cytometry experimental procedures so that data analysis can be performed in an interlaboratory setting, on multiple instruments and over prolonged periods of time. Various aspects of reproducibility and robustness will be discussed. Since development of standardized flow cytometry tests is essential for meaningful computational analyses of the data, the new possibilities that are just opening will be discussed.

    Formats Available: Streaming
    Original Seminar Date: November 12, 2014
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Standardization of flow cytometry assays: What for? How? Where does it lead us?
    Collapse Growing a Successful & Fruitful Core: You Reap What You Sow

    Joanne Lannigan has been actively involved in Flow Cytometry for over 30 years. She served as the Associate Technical Director of the Clinical Immunology and Flow Cytometry Laboratories at University Hospital at the State University of New York at Stony Brook from 1986-1999 where she supervised the clinical immunology and flow cytometry patient testing services. In 2002 she joined the Faculty of the Department of Microbiology, Immunology and Cancer Biology in the School of Medicine at the University of Virginia as a Senior Research Scientist and Director of the Flow Cytometry Core Facility. She has been an active member of ISAC since 1996 and is currently an ISAC Council member, Chair of the Shared Resource Laboratory Task Force, member of the Shared Resource Laboratory Services Oversight Committee, and Vice Chair of the Certification Advisory Committee. Ms. Lannigan's current interests involve advancing technologies in cytometry, instrument evaluation and quality assurance, flow cytometry education and core management.

    The keys to a successful research core facility are a shared responsibility between the institutional research administration and the individual core directors. Both play important roles in providing the necessary resources and strategies to insure the research missions of the cores are efficiently and effectively met. This requires the alignment of goals and coordination of efforts in business planning, forecasting, and marketing. The presenter will share some personal experiences and approaches to the every day challenges of growing a successful core. We will address such challenges as:

    • Developing the right core environment
    • The Customer-Core relationship
    • Expense recovery vs. affordable rates
    • Growing the user base
    • Establishing an efficient (lean) operation
    • Recruiting/retaining quality staff
    • Ensuring quality data
    • Planning for the future

    Approaching these challenges from a day-to-day perspective is an important part of effectively integrating the larger goals of the institution; no one knows your strengths and weaknesses better than you.

    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: February 24, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Growing a Successful & Fruitful Core: You Reap What You Sow
    Collapse The Use of Nanoparticles for Cell Identification and Tracking

    Paul Rees

    Paul Rees received his degree and Ph.D. in physics from Cardiff University in 1990 and 1993, respectively. Research for his PhD included the calculation of the optical properties of semiconductor lasers and the measurement of spontaneous emission from laser diodes under operating conditions. From 1994 to 1996, he was a Research Fellow at the Department of Physics, Trinity College, Dublin, Ireland, where his research included the study of self-pulsation in laser diodes and the theory of many-body effects in wide band-gap semiconductors. In 1996, he joined the School of Informatics, University of Wales, Bangor, U.K., where he was appointed Senior Lecturer in 2000. Research interests at Bangor included the effect of optical feedback in laser diodes, the use of chaos for data encryption, quantum computing and the theory of quantum-dot lasers.

    In 2005 he was appointed to a Chair of Nanotechnology in the newly-formed Multidisciplinary Nanotechnology Centre at Swansea University. Current research interests include the theory of the electrical and optical properties of self assembled and colloidal semiconductor quantum dots. Recent research has involved the modelling of biological cell cycles and the use of these theories in deconvolving useful information from high throughput measurement techniques such as flow cytometry and time lapse microscopy. A major focus of current research is the investigation of the uptake of nano and micro-particles in biological cells. Using the high throughput measurement techniques to investigate the heterogeneity of this uptake for large cell populations, allowing the calculation of the statistical distributions for the number particles delivered to a cell. Recent research has involved the application of engineering systems approaches to both biology and medicine in order to deconvolve meaningful information from data.

    Webinar Summary: Colloidal quantum dots are widely used as fluorescent markers for biological cells.  These particles are optically stable and bio-compatible with the cellular environment. The wavelength of the quantum dot fluorescence can be tuned across the optical spectrum by varying the size, structure and chemical composition of the colloidal particles. Initially, colloidal dots were used as direct replacements of existing fluorescence probes but more recently applications have emerged that exploit the unique properties of these nanoparticles.

    In this presentation, Paul will discuss the use of colloidal nanoparticles to measure the proliferation of cells using flow cytometry. Then the use of nanoparticles to track the movement of cells using a fluorescence microscope will be presented and finally he will outline the use of multi-colored nanoparticles to uniquely bar code cells for identification and tracking.

    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: April 16, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information The Use of Nanoparticles for Cell Identification and Tracking
    Collapse Predicting the best resolution and sensitivity in panel development and reducing inter-instrument variability in Flow Cytometry

    Presented by Steve Perfetto and Jim Wood. Moderated by Pratip Chattopadhyay

     Steve Perfetto

    Stephen P. received a B.S. degree in Medical Technology from the West Virginia University in 1977 and completed his M.S. degree in 1981 from West Virginia University. He studied and worked in the clinical Blood Bank and clinical immunology laboratories until 1988, when he joined the EPICS Division of Coulter Corporation. In 1990, he was recruited to the Walter Reed Army Institute of Research and was the manager of the core flow facility. While at WRAIR, he was involved in large HIV vaccine trials and developed functional to study the immune system of infected individuals.

    In 2000 he joined the NIH in the department of the Vaccine Research Center (VRC) as a staff scientist and manager of the flow core facility. This facility is the world-leader in multicolor flow cytometry and continues to actively develop this technology on a number of different fronts; one focus is on hardware development and reagent and analysis development. For several years, this lab has collaborated to develop Quantum Dots for use in immunophenotyping experiments. The advent of these fluorochromes provided an enormous advance in multicolor technology, allowing us to proceed from 12-color to 18-color very quickly. This group is actively working on new data analysis techniques - one major focus is the analysis and presentation of meta-data (for example, summarizing our functional analysis in which they have broken down each single response into hundreds of categories defined by the expression patterns of individual cytokines or other functional measurements). Recently, the Flow Cytometry Core broke the 18 color detection barrier by the discovery of 30 parameter flow cytometry and soon this will be further advanced to the world’s first 40 parameter instrument. Another focus is on particular aspects of immune function or viral dynamics, within the context of the major research efforts. (i) Detailed assays to an in vivo system to explore the role of CTL in lymphocyte dynamics during viral replication. (ii) Identification of viral reservoirs in CD4 T cell subsets. Sequence analysis of viruses isolated from specific CD4 T cell subsets give us an understanding of the spread of virus through the CD4 compartment, and the contribution of different CD4 subsets to both active viral production and latent reservoirs. (iii) Analysis and sorting of HIV Ag specific B cells to study neutralizing antibodies and to correlate the immunophenotyping of these cells using index sorting.

    James Wood

     

    James Wood obtained his BA in Physics from Gettysburg College; however, he also availed himself to a broad range of general and advanced biology, and chemistry courses during his undergraduate years.  He completed both his MS and PhD degrees in Biophysics from The Pennsylvania State University studying the effects on the life cycle of cells after radiation exposure.  In graduate school, he also developed the first microprocessor based two-parameter flow cytometer data acquisition system.  After completing a postgraduate position in the lab of Dr. Leon Wheeless at the University of Rochester Medical Center where he contributed to the development of a 3D slit-scan flow cytometer, he moved to Florida accepting a position with the EPICS Division of Coulter Corporation.  He was manager of the New Products Research and Applications Laboratory during most of his tenure at Coulter and Beckman-Coulter.  He currently consults for flow cytometry and pharmaceutical companies, and manages the flow cytometry shared resource at Wake Forest University School of Medicine Comprehensive Cancer Center (WFUCCC). 

    At the WFUCCC, he serves as a resource to the researchers and students.  He helps to design experiments and give advice on data interpretation.  He has worked to make flow cytometry an easily accessible technology for the Cancer Center users.  His knowledge of the technology as well as the associated biology of the applications enables him to guide the researchers and students in the successful application of the technology by explaining how to best probe the biological application with the flow cytometry technology.  He has a particular interest in the application of flow cytometry to the study of cell cycle kinetics and pursues this as time permits.  He enjoys sharing his passion for the flow cytometry technology and encouraging others to use it to its fullest extent.

    He is a long time member and contributor to International Society for the Advancement of Cytometry (ISAC) and, in the past, has served as chairperson and continues to be a member of the ISAC Data File Standards Task Force.  He has engaged in the research and development of the optical, fluidic, digital and analog electronics, and mechanical components of flow cytometers and cell sorters.  He has developed mathematical models to predict instrument performance, and has used the models to identify the critical features of a flow cytometer that limit the instrumentation’s performance. He has participated in the development of data analysis software for flow cytometry data and has worked on the development of mathematical models for cell life cycle analysis.  Currently, his interests have centered on (i) how to improve the instrument’s sensitivity and resolution of dim populations, (ii) optimizing the instrument’s ability to detect emissions from multiple fluorescent dyes, (iii) standardization of instrumentation setup and (iv) methods of data presentation and analysis. For these and other applications, he has been called upon as an expert resource to speak at users’ meetings and international courses.  He is actively developing standards for characterizing flow cytometer performance and is currently developing an LED based technology to optimize and standardize flow cytometry instrument setup.

    ---------------------------------

    Successful quantitative flow cytometry requires an understanding of the characteristics of a flow cytometer instrument that affect the measurement and analysis of the acquired photometric data.  Flow cytometer instruments must be characterized before being used for critical work.  The immediate goals include achieving better reproducibility of data, reducing variation and to facilitate intra/inter-lab comparisons of cytometry data.  Characterization  includes the determination of the instrument linearity, dynamic range, precision, accuracy, detection efficiency (Q), and electronic and optical noise (B).  In particular, Q and B affect how well cellular receptors with low expression can be measured.  For example, staining panels may be inadvertantly designed to avoid measuring dim markers on PMTs with poor resolution.  Unfortunately, current methods using bead sets to assess PMT resolution only provide rough estimates of Q and B.  Thus, staining panel design and similar protocols remain a largely empirical process, requiring time and intimate experience with an instrument’s performance. Ascertaining these instrument characteristics are the first steps toward assessing how instruments can be standardized and calibrated. Ultimately, this may be part of the validation process to certify that a flow cytometer used for critical clinical and/or GMP work is performing at the required level needed to obtain accurate and precise results.  Additionally, as part of this process we need to move from the common use of “relative intensity units (channel numbers)” to data measured and presented in physically relavent optical units (photons, photoelectrons, dye molecules, antibody binding sites).

    Recently, James Wood (Wake Forest University) developed a new device, known as an LED Pulser, to measure B and Q directly and accurately.  This device delivers consistent, uniform broad-spectrum light pulses, the amplitude of which can be adjusted with an electronic and/or an optical attenuator.  Our presentation will demonstrate how this device is used to calculate Q and B values.  In the past data from multilevel bead sets have been used to facilitate comparisons of instrument sensitivity with Q and B; however, it has proved difficult to manufacture bead sets that have closely matched intrinsic CVs.  In practice, the LED Pulser, along with calibrated fluorochrome-loaded beads, can used to determine the relationship between fluorescence channel numbers and the number of photoelectrons generated at the PMT photocathode i.e. the statistical photoelectron estimate (Spe) using a weighted quadratic fitting of pulsed LED series data.  This information, along with estimates of receptor density, can be used to calculate index values that provide quantitative guidance for panel design.  Our presentation will describe this process step-by-step, with examples.  Additionally, we will show how the metrics can be used for inter-laboratory comparisons.

    The first part of the webinar will present the principles behind the LED Pulser.  The appropriate use of the LED Pulser and how it compares to using multilevel bead sets for the calculation of Q and B. 

    The second part of the webinar will introduce how the LED Pulser, along with calibrated fluorochrome-loaded beads lead to better estimates of the four facets of predictive panel design (i) accurate Q and B values for each detector, (ii) optimal voltage settings, (iii) spillover/spreading error calculations, and (iv) estimates of receptor density and contribute to more quantitative determinations for optimizing panel design.

    -----------------------------

    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: April 28, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Predicting the best resolution and sensitivity in panel development and reducing inter-instrument variability in Flow Cytometry
    Collapse  Business Continuity and Risk Mitigation for Shared Resource Core Laboratories by Sheenah Mische

    Sheenah Mische

    Sheenah Mische, PhD, is the Senior Director in the Office of Collaborative Science. Dr. Mische oversees the Cores, providing a centralized administrative structure, and promoting state-of-the-art, service-oriented resources for the research community. Under the leadership of Associate Dean for Collaborative Science David Levy, she determines priorities for investment in new technologies, Cores, and services at the NYU Langone Medical Center. Dr. Mische has broad experience in the operation and administration of multi-technology, multidisciplinary resource centers in support of institutional research missions, both in academia and in industry.  In her most recent positions, she was Director of Translational Sciences, and Associate Director for Talent Acquisition and Academic Relations at Boehringer Ingelheim Pharmaceuticals. She was also previously Director of the Protein / DNA Technology Center at Rockefeller University.

    In recent years, research institutions and biomedical centers have suffered significant losses to research due to severe weather and other disasters.  Unfortunately, severe weather events are becoming more commonplace and Superstorm Sandy has brought climate impacts and risk into the conversation in ways not even contemplated before this October 2012 event.   The buzz word “business continuity planning” is a strategic plan to prevent, if possible, and to minimize and manage the consequences of an event that interrupts critical business processes.  From a biomedical research enterprise perspective, the business of research increasingly relies on centralized resource centers/cores, making them a critical and highly vulnerable component due to the concentration of instrumentation and resources.  This session will discuss business continuity planning, and both strategic and tactical considerations for emergency planning and risk mitigation.

    -----------------------

    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: May 27, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information  Business Continuity and Risk Mitigation for Shared Resource Core Laboratories by Sheenah Mische
    Collapse Imaging Life at High Spatiotemporal Resolution

    After obtaining a BS in Physics from Caltech, Eric Betzig moved to Cornell, where his thesis involved the development of near-field optics -- the first method to break the diffraction barrier in light microscopy. Betzig became a PI at AT&T Bell Labs in Murray Hill, NJ, where he further refined the technology and explored many applications, including high density data storage, semiconductor spectroscopy, and superresolution fluorescence imaging of cells. In 1993, Betzig was the first to image single fluorescent molecules under ambient conditions, and determine their positions to better than 1/40 of the wavelength of light. Tiring of academia, he served as Vice President of R&D at his father’s machine tool company, developing a high speed motion control technology based on an electrohydraulic hybrid drive with adaptive control algorithms. The commercial failure of the technology left him unemployed and his search for new directions culminated in the invention and demonstration of the superresolution technique PALM by himself and fellow Bell Labs expatriate, Harald Hess. Since 2005, Betzig has been a Group Leader at the Howard Hughes Medical Institute’s Janelia Farm Research Campus, developing new optical imaging technologies for biology. Betzig won the 2014 Nobel Prize in Chemistry for “The development of super-resolved fluorescence microscopy.

    Formats Available: Streaming

    Approved Credit:
  • ASCP: 1 hour CMLE
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information Imaging Life at High Spatiotemporal Resolution
    Collapse Kinetic Image Cytometry of Stem Cell-Derived Cardiomyocytes and Neurons

    Patrick McDonough has been interested in microscopy and cell biology, through out his career.    Pat’s initial interests were focused on marine organisms, and he received his Ph.D., in Marine Biology from Scripps Institution of Oceanography, in 1984.  Realizing that his true passion was experimental biology, Pat switched to research on cell biology/physiology, joining the UCSD School of Medicine, Department of Pharmacology, as a post-doctoral fellow.  Pat has always been fascinated with the physiology of cardiac and skeletal muscle cells, and the role of intracellular calcium in contraction and gene expression.  Since the late 1980’s Pat has used fluorescent calcium indicators (e.g., fluo-4, invented by Roger Tsien), to visualize and quantify calcium transients that underlie cellular phenomenon in cardiac myocytes, skeletal muscle, neurons, and other cell types.  The evolution of digital cameras, computers, automated microscopes and image analysis algorithms, set the stage for development of high throughput methods for visualizing calcium and voltage transients in excitable cells.  Pat joined Vala Sciences Inc in 2004 (founded by Jeffrey Price, a pioneer in the automated microscopy and “high content analysis”), to develop cell-based assays for drug-discovery and toxicology.  With funding from NIH STTR/SBIR grants, Vala Sciences, in collaboration with academic researchers at the Sanford Burnham Prebys Medical Discovery Institute, developed the Kinetic Image Cytometer (KIC), a purpose-built instrument to quantify calcium or voltage transients, for cells plated in 96- or 384-well formats.  Pats current research develops KIC methods to enable high-throughput screening of chemical compounds/genomic constructs for beneficial or harmful effects relevant to heart failure, arrhythmias, and neurobiology.   

    Webinar Summary:

    Kinetic image cytometry (KIC) creates single cell measurements of dynamics of events such as action potentials and calcium transients in all cells, and is especially compelling for fast-acting cells such as cardiomyocytes and neurons. Kinetic plate readers (e.g., FLIPR and FDSS) provide well-averaged readouts that require synchronized cellular behaviors, whereas our novel KIC parameterizes Ca2+ transients and action potentials on each cell; e.g., typically on 300-500 cardiomyocytes in a field of view simultaneously. Developed collaboratively by Vala Sciences and Jeff Price’s and Mark Mercola’s academic labs at Sanford-Burnham-Prebys Medical Discovery Institute (SBP), KIC has been used to detect arrhythmogenic and toxic effects in early drug discovery, and for drug discovery for heart failure. The presentation will primarily report on the accuracy of KIC for predicting arrhythmia from kinetics of the calcium transients in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), and will also include some examples of application to hiPSC-derived neurons.

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: July 28, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ASCP: 1 hour CMLE
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information Kinetic Image Cytometry of Stem Cell-Derived Cardiomyocytes and Neurons
    Collapse RNA Flow Cytometry by Steven McClellan and Paul Wallace

    Steve McClellanSteven McClellan, Flow Cytometry Core Laboratory, Mitchell Cancer Institute, University of South Alabama, Mobile, AL.

    Steve McClellan, BS, MT(ASCP) is a life science professional with over 25 years of expertise in advanced flow cytometry & cell sorting, as well as basic and clinical research in the areas of cancer biology, stem cell therapy, transplant immunology and xenotransplantation. He is currently Manager of Basic & Translational Research Operations at the University of South Alabama Mitchell Cancer Institute, where he also serves as Chief, Flow Cytometry & Imaging Core Laboratories. For the past seven years, his lab has been conducting research on cancer stem cells; working to develop better methods of purification, culture and analysis at both genetic and functional levels, as well as using cancer stem cells in HTS drug discovery.

     

    Paul WallacePaul K. Wallace, Department of Flow & Image Cytometry, Roswell Park Cancer Institute, Buffalo, NY

    Paul K. Wallace, PhD has served since 2003 as Director of the Flow and Image Cytometry Department and is Professor of Oncology at Roswell Park Cancer Institute (RPCI) in Buffalo, NY. He is also Associate Professor of Microbiology & Immunology, Dartmouth College, Hanover, NH and Associate Professor of Biotechnical and Clinical Laboratory Sciences, University at Buffalo, Buffalo, NY. He is President elect of the International Society for Advancement of Cytometry and a Councilor of the International Clinical Cytometry Society.

    Under his direction, the Flow and Image Cytometry Department at RPCI offers a strong combination of clinical and research missions.  The department’s clinical emphasis is on the diagnosis and monitoring of patients with leukemia and lymphoma.  In addition, it serves as a core reference facility performing immunophenotyping and immune monitoring studies on samples from patients enrolled in clinical trials for several biotech and grant-funded organizations.  The department’s research focus is on myeloid cell biology and translational research utilizing flow cytometry.

    Before joining RPCI, Dr. Wallace was an Assistant Professor of Immunology at Dartmouth Medical School, Hanover NH (1993-2003), a cofounder of Zynaxis Cell Science, Inc., Malvern PA (1988-1991), and the Supervisor of Flow Cytometry at SmithKline (now Quest) Clinical Laboratories King of Prussia, PA (SKCL; 1979-1988).  He is internationally recognized for his commitment to flow cytometric education and has been a member of ISAC’s Educational Task Force/Committee since its inception in 2006 and of the ICCS Education committee since 2003. He is a consultant with ASCP and CDC’s PETFAR (U.S. President's Emergency Plan for AIDS Relief), for which he has developed and presented CD4 training programs in Nigeria, India, Mozambique and Vietnam. Since 1994 he has also been on the faculty of the Bowdoin/New Mexico Annual Course in Methods and Applications of Cytometry.

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    Webinar Summary: 

    Flow cytometry permits the simultaneous measurements of many biomarkers in individual cells from bulk populations. Until now analysis has been limited, however to primarily analysis of proteins and total DNA or highly abundant DNA sequences. Since most RNA gene transcripts are present at very low quantities our ability to detect these mRNA species by flow cytometry has been limited. In 1993, Patterson et. al. (Science, 1993. 260:976) used a PCR driven in situ hybridization technique to detect HIV mRNA in infected cells, however the stringent conditions required by this technique prevented its wide spread application to and development for flow cytometry. Two new techniques, PrimeFlowTM (Affymetrix/eBioscience) and SmartFlareTM (EMD Millipore) have recently been commercially introduced and are seeing increasing use among research scientists.

    The PrimeFlow RNA assay system allows for the determination of differential RNA expression within a mixed population of cells. The assay improves sensitivity and lowers background of fluorescent in situ hybridization (FISH) through the use of a branched DNA signal amplification. In contrast, the SmartFlareTM system employs a gold nanoparticle that is actively endocytosed into most cell types. In the cytoplasm, interaction with the specific mRNA target causes the generation of a fluorescent signal, which can be detected by flow cytometry or microscopy. Protocols and data will be presented showing the usefulness of both systems. By the end of the webinar, participants should have an understanding of which technique is most appropriate for their applications; how to apply them to their research and most importantly gain an understanding of the pitfalls and solutions from individuals who have learned these the hard way.

    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit. ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: August 26, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ASCP: 1 hour CMLE
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information RNA Flow Cytometry by Steven McClellan and Paul Wallace
    Collapse Slide-Free Histology via MUSE: UV Surface Excitation Microscopy for Imaging Unsectioned Tissue by Richard Levenson

    Richard Levenson, MD, FCAP, is Professor and Vice Chair for Strategic Technologies in the Department of Pathology and Laboratory Medicine, UC Davis. He trained in medicine at University of Michigan and pathology at Washington University, and is Board-certified in Anatomic Pathology. A faculty position at Duke was followed by an appointment at Carnegie Mellon University where he helped develop multispectral imaging approaches for pathology and biology. In 1999, he joined CRI to become VP of Research, and served as Principal Investigator on federally funded research to develop multispectral microscopy systems and software for molecular pathology and diagnostics, three-dimensional small-animal imaging, optical dynamic contrast techniques, and birefringence microscopy. He serves on NIH, NCI and NSF review panels, is associate editor of Analytical Cellular Pathology, section editor for Archives of Pathology, and is on the editorial boards of Laboratory Investigation, Cytometry Part A, and Oncopathology. 

    Webinar Summary:

    Slide-free methods for rapid tissue histological analysis can cut hours off usual pathology slide preparation procedures. One approach to accomplish this is MUSE (Microscopy with UV Surface Excitation), which exploits shallow penetration of UV light to excite fluorescent signals from only the most superficial tissue elements. The method is non-destructive, and eliminates the need for conventional histology processing, formalin fixation, paraffin embedding, or thin sectioning. It requires no lasers, confocal, multiphoton or optical coherence tomography optics. MUSE generates diagnostic-quality histological images from fresh or fixed, but unsectioned tissue that can resemble conventional hematoxylin- and eosin-staining, with enhanced topographical information, and can do this rapidly, simply and inexpensively.

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit. ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming
    Original Seminar Date: September 22, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Slide-Free Histology via MUSE: UV Surface Excitation Microscopy for Imaging Unsectioned Tissue by Richard Levenson
    Collapse Optimizing SRL Performance – 1: Boost your Cell Sorting Capacity by Rui Gardner

    Rui Gardner, Gulbenkian Institute, Portugal

    Rui Gardner’s scientific career began just before graduating in Biochemistry in 1997, as a trainee in mathematical biology. This set the ground for his doctoral work, a mathematical approach to understand the prooxidative mechanisms of superoxide dismutase, paradoxically known as one of the most powerful physiological antioxidant enzymes. Most of the work was carried out in the department of Microbiology and Immunology at the University of Michigan, but also at the University of Southern California and the Gulbenkian Institute of Science in Portugal. In 2004, Rui earned his PhD in Biomedical Sciences, followed by postdoctoral work on evaluating immune diversity estimation techniques, still at the Gulbenkian Institute, where in the end of 2006 he became SRL Manager of the Flow Cytometry facility, job he still holds. Since 2007, Rui has been actively involved with ISAC’s SRL Task Force, and in 2012 was elected for ISAC’s Council, currently chairing the ISAC SRL Oversight Committee responsible for implementing and improving ISAC’s SRL-related activities and programs. In the last years, Rui has focused his efforts in addressing cell sorting performance based on the almost 10-year long experience running a cell sorting facility, as well as promoting meetings and discussions within the SRL community to help improve facility operations and management.  

     

    Webinar Summary:

    In the first of a series of webinars addressing strategies to optimize operations and management in a Shared Resource Lab (SRL) we’ll focus on enhancing cell sorting capacity. Many sorting facilities struggle with the increased demand in usage. The typically limited installed capacity of droplet cell sorters, which is directly associated to the requirement of a dedicated operator, usually leads to a narrow range of solutions to increase this capacity. Strategies are most often reduced to buying a new instrument, implying an increase in the number of FTEs or most likely increasing dramatically the burden of the current FTE, inevitably decreasing quality or SRL performance. With the advent of the new and more automated cell sorters, different and more creative solutions to increase cell sorting capacity are being addressed, including self-service sorting with all the implications associated to it. We’ll discuss several of these strategies, and present additional approaches to optimize usage of the current instrumentation, independent of their level of automation. These may include small technical implementations to reduce instrument setup, instrument troubleshooting guides, booking strategies to optimize resource usage, staggering shifts or “on call” staff to assist sorts that cannot be scheduled within normal sorting hours. There is no general strategy that will fit all facilities as some of these approaches may even collide with institutional or country-specific policies. Nevertheless, our aim is to lay down some of the possible strategies, open a discussion towards addressing this issue in a systematic way, and hopefully inspire new creative solutions from peers.

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit. ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: October 29, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Optimizing SRL Performance – 1: Boost your Cell Sorting Capacity by Rui Gardner
    Collapse Flow Cytometric Analysis of Endothelial Colony Forming Cells and Hematopoietic Progenitor Cells in Lung Vascular Disease by Kewal Asosingh and Imaging Flow Cytometry in the Study of Immune Cell Functions by Andrew Filby

    Kewal Asosingh

    Flow Cytometric Analysis of Endothelial Colony Forming Cells and Hematopoietic Progenitor Cells in Lung Vascular Disease presented by Kewal Asosingh (ISAC Scholar)

    Dr. Asosingh is a Staff Scientist, Assistant Professor of Molecular Medicine and Scientific Director for Flow Cytometry at Cleveland Clinic’s Lerner Research Institute. In 2011, he was chosen by the International Society for Advancement of Cytometry (ISAC) as an ISAC Scholar, recognition of emerging scientific research leaders in the profession. He has provided Stem Cell Cytometry training seminars at ISAC meetings, chaired oral presentation sessions on personalized medicine, and has been serving as abstract reviewer, member of the program committee for the annual ISAC meetings (“CYTO”) and reviewer for Cytometry Part A. Dr. Asosingh is also co-leader of the CYTO University (CYTO U) eLearning Delivery Task Force. Dr. Asosingh teaches flow cytometry to undergraduate and graduate students at the Cleveland Clinic and at Lakeland Community College. As Scientific Director of the Flow Cytometry Core, he serves as primary contact with investigators to provide advice on experimental design and data analysis/interpretation and assures that data generated by the Flow Core is of the highest quality.

    Webinar Summary:

    Dr. Asosingh’s lab studies bone marrow stem cells in pulmonary arterial hypertension (PAH) and pathological angiogenesis in asthma. The group has established mouse models for both diseases, and close collaboration with clinicians facilitates rapid transition of the basic research findings to clinical studies. In the past decade it has become evident that bone marrow hematopoietic stem cells exert functions beyond hematopoiesis. During development, endothelial cells and hematopoietic stem cells originate from a common bipotent mesodermal stem cell called the hemangioblast. In post-natal life, this connection is maintained via paracrine interactions between the endothelium and circulating hematopoietic progenitor cells. Their critical role in vascular homeostasis and repair of endothelial injury has opened new therapeutic perspectives for vascular degenerative diseases. The flipside of the coin is that abnormalities in hematopoiesis may underlie pathological angiogenesis, and this is exactly what his group is focused on.  Recent findings from the group show that hematopoietic stem cells isolated from the bone marrow of PAH patients are able to transfer the disease in humanized NOD SCID mice. PAH is a devastating disease characterized by endothelial cell injury, in situ thrombi and right ventricular hypertrophy. Animals engrafted with hematopoietic stem cells from PAH patients, but not from healthy control participants, had increased mobilization of progenitor cells in the peripheral blood, just like in patients, and strikingly developed many features of PAH.  Current focus of the team is to further reveal the mechanisms by which hematopoietic stem cells cause pulmonary vascular disease. This webinar will provide an overview of the different types of “endothelial progenitor cells” with special emphasis on endothelial colony forming cells and pro-angiogenic hematopoietic progenitor cells and their roles in PAH.

    Andrew Filby

    Imaging Flow Cytometry in the Study of Immune Cell Functions presented by Andrew Filby (SRL Emerging Leader)

    Dr Filby is currently head of the Flow Cytometry Core Facility at Newcastle University.  He leads a dedicated team of flow cytometry specialists with the sole aim of providing a comprehensive, cutting edge cytometry resource to the wider research community at Newcastle University and beyond.  A significant part of his focus is the development of novel cytometry-based techniques that have underpinned several high profile publications including in Science, Cell and the Cytometry Part A paper of the year accolade (2011).  He specialises in Imaging Flow Cytometry and the use of fluorescence dyes to track cell proliferation.  Prior to and post selection as an ISAC Shared Resource Laboratory Emerging Leader (SRLEL), Dr Filby has made several contributions to the Cyto conference programmes including interactive workshops and scientific talks.  He continues to contribute to the efforts of ISAC including serving on the Image Cytometry and E-learning task forces.  In addition to his work within ISAC, he is also heavily involved in supporting cytometry research and education in South America including initiatives for remote support and training.

    Webinar Summary:

    In this short webinar Dr Filby will give a brief overview of his work within the field of Imaging Flow Cytometry and what impact this has had on a number of key publications in the field of immunology and cell cycle biology.  These include the study of asymmetric cell division in the immune system, the measurement of organelle-specific calcium mobilisation in activated T cells and the analysis of the cell cycle in fission yeast.  In each case, the ability to capture and analyse literally thousands of multispectral, spatially registered images was essential to questions each method was designed to address.  Finally, Dr Filby will also discuss what the SRL EL programme means to him and how he hopes it will benefit the wider cytometry community through various initiatives he is involved in.

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit. ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: November 16, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ASCP: 1 hour CMLE
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information Flow Cytometric Analysis of Endothelial Colony Forming Cells and Hematopoietic Progenitor Cells in Lung Vascular Disease by Kewal Asosingh and Imaging Flow Cytometry in the Study of Immune Cell Functions by Andrew Filby
    Collapse Bridging Flow Cytometry with New Technologies by Monica DeLay and Sherry Thornton. Moderated by Bridget Mclaughlin

    Monica DeLay

    Monica DeLay, MS

    After receiving her master’s degree in biology, she performed research for over a decade in the field of autoimmune diseases focusing on the structure and function of human MHC and its role in modulating disease.  Throughout that time she used flow cytometry for her research and in 2008 began managing the Research Flow Cytometry Core at Cincinnati Children’s Hospital in Cincinnati, Ohio.  Over the last seven years she has helped facilitate the growth of the core in instrumentation, infrastructure, staff and clientele.  She is co-founder and president of the Ohio River Valley Cytometry Association, a local organization connecting advancements in cytometry technology to local scientists.  She is co-chair of the ABRF Flow Cytometry Research Group whose mission is to understand alterations in cell function and develop best practices associated with cell sorting.  She is a member of the organizing committee for the Core Managers workshop for the Great Lakes International Imaging and Flow Cytometry Association (GLIIFCA) and serves as a council member for the International Society for Advancement of Cytometry (ISAC).

    Sherry Thornton

    Sherry Thornton, Ph.D.

    Dr. Thornton received her Ph.D. in Developmental Biology from the University of Cincinnati College of Medicine. Her postdoctoral fellowship involved the use of flow cytometry in antigen-specific responses in the laboratory of Dr. Raphael Hirsch. She then joined the faculty in the Division of Rheumatology at Cincinnati Children’s Hospital Medical Center (CCHMC) and became flow core director for the Rheumatology Center Grant (NIH P30 AR47363). She has been extensively involved with the correlation of gene expression profiles with cell types (determined by flow cytometry analysis) of PBMC from large cohorts of Juvenile Idiopathic Arthritis patients (>500) and control specimens. Her research interests also include examination of cell types involved in the pathogenesis of arthritis. In June of 2008, Dr. Thornton led the consolidation of flow cytometry at CCHMC, and she became Director of the Research Flow Cytometry Core that houses all flow cytometry instrumentation involved in basic research at CCHMC. Dr. Thornton is active locally, nationally and internationally in the flow cytometry community. She is an active member of the International Society for Advancement of Cytometry, a founding member of the Ohio River Valley Cytometry Association, steering committee member of the Great Lakes International Imaging and Flow Cytometry Association and a member of the Career Development Committee of the Association of Biomolecular Resource Facilities.

    Webinar Summary:

    Novel applications, such as single-cell RNASeq, induced pluripotent stem cell (iPSC) generation and CRISPR-mediated genome editing, have revolutionized the way that cells can be analyzed and manipulated to investigate developmental pathways and disease processes.  These technologies can also benefit from using flow cytometry as an upstream or downstream process and, when combined, may require the involvement of several Shared Resource Labs (SRLs) from sample collection to data analysis. This webinar will highlight research projects that have utilized the above applications in combination with flow cytometry and/or cell sorting, discuss critical parameters that should be considered for successful experimental outcomes and provide strategies for coordination among core facility staff and investigators.  This discussion is aimed to provide ideas on the integration of new technologies from other cores into investigator-driven projects that utilize their SRL.

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: December 16, 2015
    On-Demand Release Date: Available Now

    Approved Credit:
  • ASCP: 1 hour CMLE
  • ISAC: 1 hour ICCE

  • Topics & Pricing InformationTopics & Pricing Information Bridging Flow Cytometry with New Technologies by Monica DeLay and Sherry Thornton. Moderated by Bridget Mclaughlin
    Collapse Beyond the Bivariate: Towards a Flow Cytometry Computational Pipeline by Wade Rogers

    Wade Rogers

    Wade Rogers has been interested in pattern discovery for about 20 years.  It began with 'TupleWare', an algorithm for discovering patterns in gene and protein sequence data, first at DuPont and later at Bristol-Myers Squibb.  Later, he and colleagues expanded to include patterns in arbitrary categorical data.  For the past 10 years he has focused almost exclusively on the challenges of analyzing flow cytometry data at the University of Pennsylvania.

    Webinar Summary:

    Single cell analyses are becoming major players in life science markets.  However, the increasing dimensionality, complexity and volume of data produced by flow cytometry pose a serious challenge that must be overcome in order to realize the full clinical potential of the method. High dimensional cell-based measurements, coupled with sophisticated computational analysis (collectively termed “cytomics”) may yield phenotypic or functional patterns that can provide informative biomarkers for discovery and clinical use.  We envision an industrial-scale computational framework that will support the development and deployment of a data analysis pipeline that (a) is robust, hardened, and fully automated, eliminating analysis subjectivity and facilitating regulatory filing, (b), can be deployed in a centralized application-as-service business model for prospective, on-demand analysis of laboratory data and (c) enables retrospective datamining of cytomic data for analysis and discovery of new biomarkers.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: February 10, 2016
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Beyond the Bivariate: Towards a Flow Cytometry Computational Pipeline by Wade Rogers
    Collapse Visualization and Analysis of Massive Scale Neuroscience Images using Vaa3D by Hanchuan Peng, Xiaoxia Liu & Zhi Zhou. Moderated by Stephen Lockett

     Hanchuan PengHanchuan Peng is a group leader at the Allen Institute for Brain Science, Seattle, USA. Inventor of a number of algorithms and software/hardware systems, including Vaa3D, BrainAligner, NeuronTracers, SmartScope, mRMR, 3D Virtual Finger, SmartACT, TeraFly, etc. A proponent for BioImage Informatics meetings, Brain Informatics conferences, journals (Bioinformatics, BMC Bioinformatics, and Brain Informatics), and BigNeuron and Open Interface of Bioimaging Software. 

     

     


    Xiaoxiao Liu is a research scientist and a Vaa3D developer at  the Allen Institute for Brain Science.  She is currently focusing on analyzing large-scale neuron morphology data and evaluating automatic neuron tracing algorithms for the BigNeuron project.  Her research interest include brain informatics,  biomedical image analysis, and high dimensional data visualization. 

     

     

     

     

    Zhi Zhou is a scientist at the Allen Institute for Brain Science, Seattle, USA. His research interest is in automatic 3D reconstruction of large-scale neurons to quantify neuronal morphology and identify cell types. He also focuses on developing algorithms and tools in order to improve the performance of large-scale image visualization and analysis.

    http://home.penglab.com/index.html

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: March 31, 2016
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Visualization and Analysis of Massive Scale Neuroscience Images using Vaa3D by Hanchuan Peng, Xiaoxia Liu & Zhi Zhou. Moderated by Stephen Lockett
    Collapse Small Instruments for Large Tasks:  High Dimensional Flow Cytometry on Compact Platforms by Bill Telford. Moderated by Paul Wallace.

    Bill Telford  Bill Telford

    • Ph.D. Michigan State University (1994)

    • Postdoc, University of Michigan (1994-1997)

    • Flow Core Director, Hospital for Special Surgery/Cornell University School of Medicine (1997-1999)

    • Flow Facility Manager and Associate Scientist, National Cancer Institute, NIH (1999-present)

    Webinar Summary:

    Modern optical and electronic technology are allowing the development of small, compact flow cytometers with the capabilities of much larger instrumentation.  We will discuss several of these cytometer platforms, and the technological advances that have made them possible.

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    This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 1 CMLE credit.  ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.

     

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: April 13, 2016
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Small Instruments for Large Tasks:  High Dimensional Flow Cytometry on Compact Platforms by Bill Telford. Moderated by Paul Wallace.
    Collapse Introduction to imaging flow cytometry and its preclinical and clinical applications

     As the Assistant Director of the Flow and Image Cytometry Facility at Roswell Park Cancer Institute, Hans Minderman oversees the research applications of the various imaging (confocal, live cell imaging, imagestream cytometry) and flow cytometry platforms available in the facility, and collaborates and consults with the investigator user base. With over 30 years of experience in clinical applications of flow and image cytometry he served as the PI on the NIH funded study on clinical application of multispectral imaging flow cytometry. Analytical approaches that were developed during this period are now actively applied in the analysis at his facility of patient samples from national and international clinical trials.  In addition, the ImageStream-specific experience gained and methods developed during this research are currently applied to support ongoing NIH- and non-NIH funded research efforts by the facility’s user base. 

     
     
    Imaging flow cytometry combines the speed, sensitivity and quantitative capabilities of conventional flow cytometry with the spatial resolution and quantitative capabilities of microscopy. In this webinar, the differences and commonalities between conventional flow cytometry and imaging flow cytometry are explained. With studies in cell signaling being one of the foremost applications of imaging flow cytometry, the audience will be guided through the analytical process of studying the NFkB signaling pathway by imaging flow cytometry. How this approach compares to conventional techniques of studying this pathway such as western blot analysis, microscopy and flow cytometry will be presented. During this exercise the basic principles of image analysis are explained  from the creation of the digitized images to how regions of interest are defined and how quantitative features can be derived. The webinar will end with a short overview of preclinical and clinical applications of imaging flow cytometry. 
     
     
    The course is intended for researchers and students who may have previous experience with conventional flow cytometry but have yet to be introduced to imaging flow cytometry. 
     
    Formats Available: On-Demand Recording, Webinar + Archive
    Original Seminar Date: May 24, 2016
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Introduction to imaging flow cytometry and its preclinical and clinical applications
    Collapse Flow Cytometry Analysis of Human Hematopoietic Progenitors in Cardiovascular Disease

    Thomas Cimato MD PhD will be presenting the webinar entitled “Flow Cytometry Analysis of Human Hematopoietic Progenitors in Cardiovascular Disease”. Dr. Cimato is an Associate Professor in the Division of Cardiology at the University at Buffalo Jacobs School of Medicine and Biomedical Sciences and Adjunct Investigator at the Roswell Park Cancer Institute.

    His research focuses on the role of inflammation on hematopoietic differentiation in atherosclerosis. He is also involved in collaborative studies that focus on the role of hematopoietic progenitor cells as immune suppressor cells in cancer.

    In this webinar he will discuss recent advances in our understanding of how early hematopoietic progenitors respond to hypercholesterolemia and myocardial infarction in animal models to accelerate atherosclerosis. He will discuss the development of an assay to measure human hematopoietic progenitors in blood, and early translational studies in human subjects with heart disease. 

    Formats Available: Streaming, Webinar + Archive
    Original Seminar Date: September 27, 2016
    On-Demand Release Date: Available Now

    Approved Credit:
  • ISAC: 1 hour ICCE
  • ASCP: 1 hour CMLE

  • Topics & Pricing InformationTopics & Pricing Information Flow Cytometry Analysis of Human Hematopoietic Progenitors in Cardiovascular Disease