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UWCCC Flow Cytometry Core Facility Issue 2, Fall 2012

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Call us at

(608) 263-0313

v MACSQuant o multi-color flow cytometry

o reduced cost

o automated staining

o automated acquisition

Instrumentation 18 parameter Cell Sorter

BD FACSAria

15 parameter benchtop flow cytometer

BD LSR II

6 parameter benchtop flow cytometer

BD FACS Calibur

High-content kinetic Imaging System

BD Pathway

Cell counter Nexcelomm

10 parameter benchtop flow cytometer with inline cell separation

MACS Quant

We are located at 7003 WIMR (office) 7016 WIMR (lab) 1111 Highland Ave Madison, WI 53706

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Fluorescent microscope

Olympus

FlowJo Site License The University of Wisconsin has acquired an institutional, shared-cost software license. Register any computer on- or off-campus for a $250 annual fee. Turn in a dongle for a credit for 4 years of use. The license ensures immediate access to any new versions of the software. see Page 2 for more

Call for group demonstration or group training!

We are expecting the arrival of a brand new ARIA III cell sorter. This sorter will be equipped with a Level II biosafety cabinet providing sterile conditions for sorts and BSL2 containment for human samples. We expect to open the Aria III for business in November!

Visit us at

www.uwhealth.org/flowlab

Join this online flow cytometry network and check out the panel design tool. Find reagents and access our core instrument configurations.

SPICE Simplistic Presentation of Incredibly Complex Evaluations

http://exon.niaid.nih.gov/spice SPICE is a data mining software to analyze and interpret highly complex multi-color flow cytometry data. It can process large polychromatic FlowJo files and organizes the normalized data graphically. It simplifies analysis of highly sophisticated assays and may aid discovery of potential new correlations.

Flow Newsletter Issue 2 / Fall 2012

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FlowJo® Site License Want to reduce the cost of FlowJo software?

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We are excited to introduce a new opportunity to greatly reduce your cost on FlowJo analysis software. The University of Wisconsin-Madison has acquired a site license, which enables the whole community to access the most recent versions of the analysis software without a dongle. Any internet connection will work, and the computer does not need to be located on campus. Any personal computer may be registered under this program. Individual FlowJo dongles may be returned for site license or flow core credit through our office. There will be no need for investing any more in dongle-upgrades, and the newest versions of FlowJo will be readily available with the license. To obtain a serial number, go to our website and follow instructions. The annual cost for the site license is expected to be around $250 this fiscal year, but we hope we may further reduce this price with recruitment of more users to this shared program. The site license ensures ready access to any new releases of the software. The site license works on Macintosh, PC or Linux operation systems. It supports any software versions from 5.6 up. It gives access to the universal Version 10 and will not require any further investment to access the upcoming Version 11, which

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will otherwise require a new-generation dongle upgrade. Version 9.x (Mac) and 7.x (PC) have many great features on its Advanced Platforms (see on Page 3) to support complex multi-color flow cytometry assays. The

Kinetics platform will outline kinetic changes by comparing a series of samples automatically and performing statistic analysis on them. The

Proliferation platforms may determine various subpopulations of interest (like CFSE fingers, etc) automatically. Complex characterization assays are supported by Cluster analysis and Boolean gating when various subpopulations are determined and compared automatically based on any potential combinations of multi-color staining (ie: to compare any single, double or triple cytokine co-production). Population Comparison and Multi-Sample Comparison are tools to expedite data analysis and statistical tests on complex assays. Multi-dimensional cell-cycle analysis and rare-event analysis enable state-of-the-art flow cytometry assay development. The Verify Sample Validity is a

quality control tool, which identifies samples with potential acquisition errors compromising the read-out (clogs, laser drifts or event rate fluctuations).

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We offer a FlowJo license credit of $1000 for every dongle returned via

our site license program.

Version 10 is out! v10 is the universal (Mac, PC and Linux compatible) preview version of the upcoming v11 edition. The re-designed, ribbon-style, custom user interface makes it very easy to navigate for even the beginner level user. The layout editor is upgraded with many extra graphic design features to enable the direct preparation of high-resolution, publication ready figures. The 3D and web-animation features add a state-of-the-art quality to flow cytometry data presentation. Please note, that this version is a ‘preview’ of future v11 and Advanced Platforms are not added back just yet.

Flow Newsletter Issue 2 / Fall 2012

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FlowJo® – Advanced Platforms State-of-the-art tools for complex, multi-color flow analysis

Concatenate. Create new FCS files after modifications in FlowJo. Merged files can be used for kinetic assays, time-points, concentration gradients, and also to increase event numbers.

Cluster Analysis. Defines subpopulations automatically. Kinetic Analysis. Mathematical modeling to calculate time-dependent data (like Ca2+-flux).

Proliferation. Mathematical determination of cell division peaks (ie: CFSE fingers).

Multigraph Overlay. Visualizes all combinations of data on selected, well-defined subpopulations.

Population Comparison. Compares any single read-outs on a defined subpopulation to characterize differences.

Flow Newsletter Issue 2 / Fall 2012

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Ask the Flow Core Staff

Core Staff Profile Have a question about flow cytometry, imaging or about our other core services? Contact us!

Manish Patankar, PhD Faculty Supervisor patankar@wisc.edu

Dagna Sheerar, BS Education, Consultation, Administration, Billing, Communication, Outreach, BD Pathway dsheerar@wisc.edu

Karen Ersland, PhD Cell Sorting, Daily Quality Control, Assay Services, Hands-on Training, Data Analysis, ModFit, Figure preparations ke@wisc.edu

Faye Bruggink Online schedule, MACSQuant Protocol Development fbruggink@wisc.edu

Erika Heninger Cell Sorting, Biosafety Protocol Amendments, Consultation, FlowJo Site License, Data Analysis, Figure preparations, Multi-color assay development eheninger@wisc.edu

“Introduction to Flow Cytometry” Lecture Series Fall / Winter schedule October 22, 24, 26 November 26, 28, 30 December - no

Flow Core NewsLetter, Issue 2/ Fall 2012 Editor: Erika Hėninger, eheninger@wisc.edu

For instrument configurations please visit our website at www.uwhealth.org/flowlab. If you have a special request or if you are working with an unusual or novel fluorochrome please contact us to see if we have the right filters for you.

Data Analysis & Consultation Need help with data analysis or guidance to navigate the myriad of new options in flow cytometry & imaging? Schedule a consultation and we will be happy to assist you with both experimental design and interpretation of data. We also offer assistance with preparation of publication-ready figures and other flow cytometry data presentations.

Core Services

Assay Services

Our facility offers standardized assay services including everything from sample processing to data interpretation. Besides the assured Quality Control and high-throughput options, this service has been providing great value to laboratories without equipment or trained staff to perform flow cytometry & imaging. Contact us to see what we can offer to develop customized protocols for your experimental models.

Training & Education

For new users we offer a monthly lecture series titled “Introduction into Flow Cytometry”. We provide one-on-one, hands-on training sessions on all of the instruments. Assisted services are also available for inexperienced users and experienced users with unfamiliar assays, on schedule or upon request. Training sessions are customized to our users’ individual experimental needs.

Instrument specifications

Manish Patankar, PhD Facility Director Dr. Patankar joined the laboratory in May of 2011. He is an Associate Professor in the Research Division of the University of Wisconsin – Madison Department of Obstetrics and Gynecology and a member of the UWCCC. Dr. Patankar’s research focuses on the development of diagnostic tests for ovarian cancer and pre-eclampsia and strategies for the treatment of ovarian cancer. He and his lab have been active users of the facility since his arrival at the University in 2004.

BioSafety Updates

Renewing your

biosafety protocol? Adding new flow experiments or sorting projects? Please contact us for updated flow cytometry SOP’s and safety guidelines to expedite your amendment process. Please note, that we are in the process of adapting state-of-the-art safety guidelines for live cell sorting including pre-screening of human samples.