CYTO 2013: Scientific Tutorials
All CYTO U courses qualify for ICCE continuing education credit. If you would like to see a list of topics included in this recording, please click on the "Topics" tab.
Seminar Information
Seminar Date:
May 18, 2013
Click on a topic name to see details and purchase options.
Saturday May 18, 2013
In the first part of the Tutorial (Telford), flow cytometric methods to assess apoptosis will be thoroughly reviewed.  Students will learn how to select the appropriate assay, the technical details necessary for assay success, and combining multiple assays for multiparametric analysis of cell.  The tutorial will take a very practical approach, and actual cytometry data will be analyzed as part of the program.  Autophagy, a phenomenon both related to and distinct from cell death will also be covered.

In the second part of the Tutorial (Darzynkiewicz), detection of DNA damage by flow and image cytometry will be covered, both as a component of apoptosis and as a means of analyzing the effects of pharmacological, toxicological and environmental insults to live cells.  Detection of both DNA damage itself and of phosphoproteins critical for DNA organization and structure that serve as markers for DNA damage will be particularly emphasized.   

After completing the Tutorial, the student should be able to:Have a working knowledge of flow cytometric assays for apoptosis, autophagy and DNA damage.

Be able to select an appropriate apoptosis and DNA damage assay for their experimental system, procure the necessary reagents and carry out the assay.
Combine multiple apoptosis assays to develop powerful systems for studying the complex signaling and progression of apoptosis

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
Bill Telford   [ view bio ]
Cytometers make optical measurements of particles. Using flow cytometers as the primary example, this tutorial will provide an understanding of the measurement process and how to critically and objectively characterize the instrument's ability to make fluorescence and light scattering measurements. The use of standard instrument performance criteria and testing materials can allow minimal performance requirements to be established for an assay and assure that results on different instruments in different laboratories get comparable results when running the same sample. Authoritative standards (traceable to a standards setting organization) are generally not available for cytometry. Students will learn about the use and limitations of commercially available standard materials and some options for laboratory prepared standards.

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
Robert Hoffman   [ view bio ]
This tutorial will provide the attendee with an overview of biosafety principles, as they apply to flow cytometry with emphasis on cell sorting. The process of risk assessment for determination of biosafety containment levels and the development of a Standard Operating Procedure (SOP) for flow cytometers will be discussed.

All CYTO U courses qualify for ICCE continuing education credit.
Speaker Information
Kevin Holmes   [ view bio ]
High throughput screening is now a real opportunity with flow cytometry. It is now possible to collect thousands of samples per day in a highly ordered way. However, designing and preparing these assays does require some careful attention to details and automated prep becomes an important component. One advantage is that you only sample very small volumes (like 1 uL) instead of 50 to 250 uL. This means instead of needing far more cells to start, you may actually be able to run far more samples than you might think. We will discuss several different implementations of HT flow cytometry and the types of assays that are usable in this format. This tutorial will also discuss aspects very high content flow cytometry where you might have a very large number of samples as well as a large number of parameters such as might be found in CyTOF data where parameters may exceed 20-30 per assay.

After participating in this tutorial, the participant will be more aware of the advantages and possible disadvantages of having to work in an automated, or semi-automated régime. They will have a good knowledge of how to approach the preparation, and the analysis of data. They will also learn some analytical tools that have been designed particularly for high throughput and high content analysis.

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
J.Paul Robinson   [ view bio ]
Image-based experiments using cultured cells have proven to be a powerful means of generating information-rich data for biological applications. This tutorial will instruct biologists in the concepts and application of CellProfiler, an open-source, freely-downloadable software package designed for large-scale, automated phenotypic image analysis. We will work through hands-on examples, including construction of an analysis pipeline, optimization of module settings, automatic cellular feature detection and measurement, and exporting results. We will also briefly discuss the basic principles of supervised machine learning in order to score phenotypes where phenotypic differences between samples are subtle and/or complex. The tutorial participant should gain a working knowledge of CellProfiler and how to process and analyze their high- or low-throughput, high-content experiment.

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
David Logan   [ view bio ]
This tutorial will cover the latest tips and tricks for successful polychromatic flow cytometry experiments. Participants will leave the session with a practical, working knowledge of how to develop or optimize them.

All CYTO U courses qualify for ICCE continuing education credit.
Speaker Information
Pratip Chattopadhyay NIAID, NIH   [ view bio ]
The use of fluorescence (Förster) resonance energy transfer (FRET) for assessing molecular interactions in cellular systems is exponentially expanding. Several methods, mostly for microscopy, have been proposed, but most of them that made it to broad use owed to their simplicity suffer from being only qualitative or even from being prone to misinterpretation of results. The educational outcome of the tutorial is to provide the audience with stable foundations for applying a simple, yet qualitative FRET procedure that can be performed in any commonly available laser scanning fluorescence microscope. Addition outcomes include skills in interpretation of FRET data, and a broader knowledge on the pros and cons of various FRET methods that allow an educated choice of the approach most appropriate for the biological question.

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
Gyorgy Vereb   [ view bio ]
Core facilities are now common in all work settings. Flow cytometry is a well-established technique but the core faces particular challenges in the face of expanding technology. In particular, cores need to bring added value to their users and institutional setting. But how can core facility staff keep up with the latest developments, how can they receive appropriate continuing education and how can this be passed on to users of a facility? We will discuss evaluation of technology and strategies for importing this into a core and also how education on site, at relevant meetings and by remote learning can benefit the facility. At the end of the tutorial the delegate will be aware of the approaches that can be taken to bring added value to the core, its staff and its users.

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
Alfonso Blanco   [ view bio ]
Derek Davies   [ view bio ]
CellOrganizer is an open source software system that can learn models of the size, shape and spatial distribution of cellular components directly from images.  These models are generative, which means that they can be used to synthesize new images of cells that are statistically similar to the ones they were trained on.  Such images are useful for testing image analysis algorithms, and can be used as the basis for spatially-realistic cell simulations using systems such as Virtual Cell and MCell.  Perhaps most importantly, CellOrganizer models represent a transportable means of representing the results of High Content Screening (HCS) assays that is not dependent on a specific instrument, assay or cell type.  

This tutorial will focus on how to use CellOrganizer and how to interface it with other software. The tutorial will begin with a brief overview of the conditional structure of the models within CellOrganizer and the system organization.  The first part of the tutorial will focus on training generative models. Students are strongly encouraged (but not required) to bring a laptop.  Attendees are also encouraged to bring a fluorescent cellular image dataset of their own to use for building a model, but datasets will be available at the tutorial for attendees who do not have one. Ideally, images should be two or three dimensional single cell images (i.e., already segmented) with different fluorescence channels for a fluorescently labeled target protein (ideally a protein showing a punctate or vescular pattern), a cell membrane or cytosolic-labeled marker, and a DNA marker (but these are not strict requirements).  

The second part will focus on synthesizing cell images from the models and importing the images or model parameters into other software systems.  The last part will focus on adding new capabilities to the open source system, such as modules for building new types of components.

Students should leave this session with mastery of the principles behind building probabilistic models from images and practical experience with training and using them with CellOrganizer. They will be able to use them to compare results from different HCS assays using the generative model parameters, and import synthetic images into cell simulation systems.

All CYTO U courses qualify for ICCE continuing education credit.

Speaker Information
Gregory Johnson   [ view bio ]
Devin Sullivan   [ view bio ]
This tutorial will address how to find the proverbial 'needle in the haystack' when needing to identify or sort rare populations of cells. In the contemporary flow cytometry laboratory, there is often the need to accurate identify rare cells, such as circulating endothelial cell, endothelial progenitor cells, tumor cells, or immune subpopulations such as plasmacytoid or monocytoid dendritic cells.The student will gain an appreciation of obstacles in the accurate identification of rare cells and of strategies to overcome these obstacles and to assure better experimental data.

All CYTO U courses qualify for ICCE continuing education credit.
Speaker Information
Phil McCoy   [ view bio ]
Individual topic purchase: Selected
International Society for Advancement of Cytometry
ICCE: 15.00
American Society for Clinical Pathology
CMLE: 15.00
This continuing medical laboratory education activity is recognized by the American Society for Clinical Pathology for 15 CMLE credit. ASCP CMLE credits are acceptable for the ASCP Board of Registry Certification Maintenance Program.
ISAC Member Price: $0.00
Non-Member Price:$375.00