HigH-Content AnAlysis

Register by November 30 th SAVE up to $200! 

Cambridge Healthtech Institute’s TENTH Annual 

High-Content 

January 8-11, 2013 

The Fairmont Hotel l San Francisco, California 

Analysis 

The most 

COMPREHENSIVE 

COVERAGE of HCA 

applications and 

technologies. 

Anniversary 

TOPICS INCLUDE: 

High-Content Screening in 1536-Well Format 

High-Content Image and Data Analysis 

HCA for Toxicity Assessment 

Novel Biosensor Assays for Screening 

Phenotypic Drug Discovery 

3-D Cell-Based Assays 

HCA of 3-D Tumor Spheroids 

Digital Pathology and Tissue Diagnostics 

HCA of Stem Cells 

RNAi Screens 

Advanced High-Content 

Analysis Course 

January 7, 2013 

User Group Meetings 


Dinner Course: Advanced Data 

Analysis for Phenotypic Screening 


Live-Cell Imaging Workshop 


Executive Sponsor 

Premier Sponsor 

Corporate Sponsors 

Organized by 

Cambridge Healthtech Institute 

HighContentAnalysis.com

Cambridge Healthtech 

Institute’s TENTH Annual 

January 8-11, 2013 

CHI offers comprehensive sponsorship 

packages which include presentation 

opportunities, exhibit space and branding, 

as well as the use of the pre- and post-show 

delegate lists. Customizable sponsorship 

packages allow you to achieve your objectives 

before, during, and long after the event. Signing 

on early will allow you to maximize exposure to 

hard-to-reach decision makers! 

Agenda Presentations 

Showcase your solutions to a guaranteed, 

highly-targeted audience. Package includes a 

15- or 30-minute podium presentation within 

the scientific agenda, exhibit space, on-site 

branding and access to cooperative marketing 

efforts by CHI. 

User Group Meetings - Sold Out! 

Co-locate your user group meeting with High- 

Content Analysis 2013. CHI will help market the 

event, manage logistical operations, develop the 

agenda, and more. CHI can handle the entirety 

of the meeting, or aspects of your choosing. 

Pre-Conference Workshops/Short Courses 

Includes a 15-minute or 30-minute podium 

presentation during pre-conference workshop, 

as well as your company logo displayed on 

pre-conference workshop materials and 

on-site signage. 

Focus Groups 

CHI will gladly provide you the opportunity of 

running a focus group on-site. This exclusive 

gathering can be useful to conduct market 

research, gather feedback on a new product 

idea and gather marketing intelligence from 

industry experts on a specific topic. 

Invitation-Only VIP Dinner/Hospitality Suite 

Sponsor will select invitees from the 

conference pre-registration list for an evening 

of networking at the hotel or a top local venue. 

CHI will extend invitations, conduct follow-up 

and monitor responses. Reminder cards will 

be placed in the badges of delegates who will 

be attending. 

Exhibit Information 

Exhibitors will enjoy facilitated networking 

opportunities with qualified decision makers at 

High-Content Analysis, making it the perfect 

platform to launch a new product, collect 

feedback and generate new leads. Exhibit space 

sells out quickly, so reserve yours today! 


Analysis 

Sponsorship & Exhibit Information 

Anniversary 

Looking for additional ways to drive leads 

to your sales team Cambridge Healthtech 

Institute can help with custom lead 

generation programs! 

We offer clients numerous options for custom 

lead generation programs to address their 

marketing and sales needs. Some of our 

programs include: 

• Live Webinars • White Papers 

• Market Surveys • Podcasts and More! 

Benefits of working with Cambridge 

Healthtech Institute for your lead generation 

needs: 

• Your campaign will receive targeted 

promotion to Cambridge Healthtech 

Institute’s unparalleled database of over 

800,000 individuals, all of which are involved 

in all sectors of the life sciences – lists can 

be segmented based on geography, research 

area, title and industry. 

• All custom lead generation programs are 

promoted through our experienced marketing 

team that will develop and drive targeted 

campaigns to drive awareness and leads to 

your lead generation program. 

• For our webinar programs, we offer 

assistance in procuring speakers for your 

web symposia through our extensive roster 

of industry-recognized speakers across 

multiple disciplines within life sciences, as 

well as provide an experienced moderator 

and dedicated operations team that will 

coordinate all efforts. 

• If choosing a white paper program, we can 

offer editorial experience and provide an 

industry-recognized author to write your 

white paper. 

To customize your participation as a sponsor 

or exhibitor at this event, please contact: 

Katelin Fitzgerald 

Manager, Business Development 

781-972-5458 | kfitzgerald@healthtech.com 

Hotel & Travel Information 

Conference Hotel: 

The Fairmont San Francisco 

950 Mason Street 

San Francisco, CA 94108 

Phone: 415-772-5000 

Discounted Room Rate: $219 s/d 

Discounted Room Rate Cut-off Date: December 

10, 2012 

Please call the hotel directly to reserve your 

sleeping accommodations. You will need to 

identify yourself as a Cambridge Healthtech 

Institute conference attendee to receive the 

discounted room rate with the host hotel. 

Reservations made after the cut-off date or after 

the group room block has been filled (whichever 

comes first) will be accepted on a space and rateavailability 

basis. Rooms are limited, so please 

book early. 

Flight Discounts: 

Special discount rentals have been established 

with American Airlines for this conference. 

• Call American Airlines 1-800-433-1790 and use 

Conference code 9413BL. 

• Go online www.aa.com and enter Conference 

code 9413BL in promotion discount box. 

• Contact our dedicated travel agents at 1-877- 

559-5549 or chi@protravelinc.com. 

Car Rental Discounts: 

Special discount rentals have been established 

with Hertz for this conference. Please use one of 

the following methods: 

• Call Hertz 1-800-654-3131 and use our Hertz 

Convention Number (CV): 04KL0003 

• Go online www.hertz.com and use our Hertz 

Convention Number (CV): 04KL0003 

Lead Sponsoring Publication 

Sponsoring Publication 

Sponsoring 

Organization 

Media 

Partners 

2 | High-Content Analysis HighContentAnalysis.com



January 8-11, 2013 


Analysis 

Anniversary 

Monday, January 7, 2013 Pre-Conference Short course 

Advanced High-Content Analysis Course* 

Beyond the State-of-the-Art in HCA 

8:30-9:00 am Registration for Advanced High-Content Analysis Course 

9:00-10:30 Part 1: The HCS/A Platforms: What’s under the Hood 

Anthony M. Davies, Ph.D., Director, Irish National Center for High-Content Screening and Analysis (INCHA) 

10:30-11:00 Coffee Break 

11:00 am-12:30 pm Part 2: Data Analysis and Management 

Karol Kozak, Ph.D., Head, Data Handling Unit and High-Content Screening, ETH Zurich 

12:30-1:30 Enjoy Lunch on Your Own 

1:30-3:00 Part 3: HCA/HCS Assay Development 

Paul A. Johnston, Ph.D., Research Associate Professor, Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh 

3:00-3:30 Refreshment Break 

3:30-5:00 Part 4: Questions and Panel Discussion 

5:00 Close of Course 

h Please visit www.HighContentAnalysis.com for a detailed course description. 

*Separate registration required 

Tuesday, January 8, 2013 User Groups 

9:00 am-12:30 pm GE Healthcare User Group Meeting 

9:00 am-2:00 pm Molecular Devices User Group Meeting 

2:00-6:00 pm PerkinElmer User Group Meeting 

2:30-7:00 pm Thermo Scientific User Group Meeting 

5:00-6:00 pm Conference Pre-Registration 

Wednesday, January 9, 2013 Main Conference 

7:30-8:30 am Conference Registration 

8:30-8:40 Welcome Remarks from Conference Director 

Julia Boguslavsky, Executive Director, Conferences, Cambridge Healthtech Institute 

8:40-8:45 Welcome Remarks from Executive Sponsor 

Scott Keefer, MBA, Manager, Product Management, Thermo Scientific High Content Products 

Sponsored by 

8:45-9:45 Opening Panel Discussion 

Tenth Anniversary of HCA: Progress Report 

Hear an update from participants of the Inaugural HCA meeting 10 years ago, including recent developments in technology 

and applications, as well as remaining unmet needs and future directions. 

Chairperson: Anthony M. Davies, Ph.D., Director, Irish National Center for High-Content Screening and Analysis (INCHA) 

Panelists: 

Paul A. Johnston, Ph.D., Research Associate Professor, Pharmaceutical Sciences, School of Pharmacy, University 

of Pittsburgh 

Jonathan A. Lee, Ph.D., Senior Research Advisor, Quantitative Biology, Eli Lilly & Co. 

Joe Trask, Ph.D., Head, Cellular Imaging Core, The Hamner Institutes for Health Sciences 

9:45-10:45 Coffee Break in the Exhibit Hall with Poster Viewing 

Anniversary 

Sponsored by 

HighContentAnalysis.com High-Content Analysis | 3



January 8-11, 2013 


Analysis 

Anniversary 

High-Content Screening in 1536-Well Format 

10:45-10:50 Chairperson’s Opening Remarks 

Richik N. Ghosh, Ph.D., Director, Research & Applications, Thermo 

Scientific High Content Products 

10:50-11:15 Developing a High-Throughput High-Content 

Infrastructure and the Impact of 1536-Well HCS on BMS 

Drug Discovery 

Debra Nickischer, Research Scientist II, Cellular Systems, Molecular 

Sciences and Candidate Optimization, Bristol-Myers Squibb 

There is a growing interest within the drug discovery industry to utilize 

more physiologically relevant cellular models in early hit-identification 

efforts, with the goal of identifying higher-quality drug candidates and 

enabling better prediction of liabilities. For high-throughput screening with 

large compound libraries, this translates to a need to enable sophisticated 

assay platforms such as high-content screening in a miniaturized, 1536- 

well microplate format. With focus on specific BMS programs, we will 

discuss considerations and challenges for developing high-throughput HCS 

capabilities and will outline innovative approaches to analyze and interpret 

the large volumes of content-rich data this platform provides. 

11:15-11:40 A Miniaturized 1536-Well HCS Pipeline to Identify 

Small Chemical Compounds Improving Muscle Function 

Enrico Schmidt, Ph.D., Lab Head and Investigator III, Center for 

Proteomic Chemistry, Integrated Lead Finding 1, Novartis Pharma AG 

11:40-11:55 New CCD Camera Technology for Sponsored by 

High-Content Screening 

Audra Ziegenfuss, Technical Product Manager, Thermo 

Scientific High Content Products 

As camera technology advances, higher sensitivity and better resolution 

are the results. We will be discussing a new CCD camera used in highcontent 

analysis and comparing it to current technologies being used in 

the high-content space. 

Sponsored by 

11:55-12:10 pm Enabling High-Throughput HCS 

with the Cell Insight 

Debra Nickischer, Research Scientist II, Cellular Systems, 

Molecular Sciences and Candidate Optimization, Bristol-Myers Squibb 

We will discuss how BMS is utilizing the Thermo Scientific CellInsight HCS 

Platform in Lead Discovery. We will describe our automation configurations 

with the Thermo Scientific Orbitor plate handlers; how the CellInsights 

have enabled our high-throughput, high-content screens; and the impact 

on BMS drug discovery. 

12:10- 12:25 New Developments in High 

Sponsored by 

Content Imaging Systems for Faster and 

More Efficient Screening 

HaiGuang Zhang, Ph.D., Senior Application 

Consultant, GE Healthcare 

Rapid acquisition without compromising on image quality is essential 

for successful high content screening, particularly in high well-density 

formats. GE Healthcare is continuing to push the boundaries of 

HCS by incorporating the latest advances in hardware and software 

technologies for optical imaging. We will discuss new components and 

features of the IN Cell Analyzer systems that are enabling more rapid, 

robust and efficient hit identification and assessment. 

12:25-12:40 Sponsored Presentation 

(Opportunity available. Contact Katelin Fitzgerald at kfitzgerald@ 

healthtech.com or 781-972-5458.) 


High-Content Image and Data Analysis 


10:50-11:15 Flat Field Correction and Multi-Parametric 

Regression Models for High-Content Analysis 

Peter Horvath, Ph.D., Data Analysis Specialist, Light Microscopy and 

Screening Center, ETH Zurich 

11:15-11:40 Content-Based Searching of Bioimage Databases 

Robert F. Murphy, Ph.D., Professor, Computational Biology and 

Biological Sciences, Biomedical Engineering and Machine Learning; 

Director, Ray and Stephanie Lane Center for Computational Biology, 

Carnegie Mellon University 

We have developed OMERO.searcher as the first of a series of open source 

tools that can augment the capabilities of a bioimage database system such 

as OMERO. OMERO.searcher Server can be installed on top of an OMERO 

database to allow both internal and external users to perform image content 

searches. These searches can be done to find images similar to a selected 

image (or images) already in the database, or using images on a user’s own 

computer. External users can also use OMERO.searcher Local Client to 

search one or more remote databases using similarity to local images, and 

searching of databases that use systems other than OMERO can also be 

easily enabled. I will discuss our experience with adapting other advanced 

image analysis tools for use with OMERO databases. 

11:40-12:05 pm Making an Individual Cell’s Voice Heard: The 

Beauty of High-Content Screening 

Tiao Xie, Ph.D., Leader, Image and Data Analysis Core (IDAC), Harvard 

Medical School 

A wide spectrum of image analysis tools/approaches have been utilized 

at IDAC to quantify the image datasets from the diverse image-based 

screening assays. The recent development of commercial image analysis 

packages has enabled us to robustly analyze data generated from the more 

traditional image-based assays such as cell viability, mitotic index and protein 

expression/translocation assays. However, some assays that target very 

specific morphological changes of whole cells or sub-cellular structures 

require more customized analysis solutions to extract specific information 

from the images. Moreover, the massive scale of image datasets generated 

from high-throughput screens also call for integrated data management 

approaches, including image storage, sharing, visualization, analysis and 

secondary data handling. Several successful stories will be presented to 

demonstrate our high-content screening capacity at Harvard, from image 

acquisition/storage, to high-content analysis and data visualization. 

12:05-12:20 Complex Challenges in the Field Sponsored by 

of Cellular Level Research Require NEW Paths 

in Your Road to Discovery 

Robert Graves, Ph.D., Senior Application Specialist, GE 

Healthcare 

For today’s highly automated systems for image acquisition requires insight 

into cells and intracellular components. The tools you need for everyday 

assays are complex, unique applications requires a comprehensive 

package of image analysis tools for a broad range of imaging experiments. 

Come and learn about our highly adaptive software configured for a range 

of skills and experience to meet your challenges in the modern lab. 

12:20-12:35 Sponsored Presentation 

(Opportunity available. Contact Katelin Fitzgerald at kfitzgerald@ 






January 8-11, 2013 


Analysis 

Anniversary 

HCA for Toxicity Assessment 

1:45-2:10 High-Content Screening of Zebrafish Embryos for 

Drug Safety Assessment 

Jyotshna Kanungo, Ph.D., Principal Investigator, Lead Scientist, Zebrafish 

HTS Laboratory, National Center for Toxicological Research, FDA 

Zebrafish embryos are being routinely used in chemical toxicity 

assessments. Since these tests require minimal amounts of test 

compounds, test durations are less time-consuming than typical toxicity 

tests, and multi-parametric high-content assays based on a variety of 

sub-lethal endpoints can easily be performed including those useful 

for the assessment of a chemical’s teratogenicity. We have developed 

several endpoints for high-content analysis of drug effects on the zebrafish 

embryos. In one approach, using multi-well plates, a high-content 

automated imaging procedure is employed to measure axon length in 

zebrafish embryos in vivo to screen for drug effects and determine dose 

levels that are developmentally toxic. Automated fluorescent image 

acquisition of the transgenic embryos is performed in vivo with embryos 

arrayed in 384-well plates, and post-acquisition image analysis provides 

average axon lengths in the control and experimental groups. 

2:10-2:35 Removing the Edge Effect Limitation of 

Cytotoxicity Testing 

Peter J. O’Brien, D.V.M., Ph.D., Veterinary Clinical Pathologist, Pathology, 

University College Dublin 

The edge effect is one of the most limiting factors in performing 

cytotoxicity tests conducted over multiple days. Microenvironments of 

wells at the edges of microtiter plates can restrict cell growth and function 

and response to treatment. This effect is also found, although to a lesser 

extent, at the wells near to the edge and found to a greater extent with 

wells at plate corners. A recently-developed microtitre plate has largely 

eliminated this adverse effect, with a proprietary method for maintaining 

a constant microenvironment across all wells. The positive impact of 

this development on sample throughput, measurement precision, and 

accuracy of cytotoxicity assessment will be demonstrated. 

2:35-3:00 Contribution of Physicochemical Properties to Toxicity 

Shuyan Lu, Principal Scientist, Drug Safety Research & Development, Pfizer 

We examined the relationship between physicochemical properties, 

such as partition coefficient (clogP), topological polar surface area (TPSA), 

acid dissociation constant (pK(a)), and in vitro mechanistic endpoints 

generated using a high-content imaging approach. We demonstrate 

in our initial analysis that compounds with clogP>2 and pK(a)>5.5 

flagged more endpoints than compounds with clogP ≤ 2 and pK(a) ≤ 

5.5. When this knowledge was applied to eight different mechanistic 

cytotoxicity endpoints (cell loss, apoptosis, ER stress, DNA fragmentation, 

mitochondrial potential, nuclear size, neutral lipids/steatosis and lysosomal 

mass), we found that compounds with such properties preferentially 

flagged in the lysosomal endpoint. We also saw a slight enrichment 

of such compounds in the endpoints cell loss, DNA fragmentation 

and nuclear size. In addition we demonstrated the contribution of 

physicochemical property to cardiotoxicity induced by imatinib. 

3:00-3:15 Cardiotoxicity of Oncology Drugs: Sponsored by 

High Content Analysis of Kinase Inhibitors in 

a Human Stem Cell Derived 

Cardiomyocyte Model 

Nick Thomas, Ph.D., Principal Scientist, Cell 

Technologies, GE Healthcare 

Industrial scale production of cardiomyocytes from human stem cells 

provides the potential to develop novel and improved physiologically 

relevant and human-predictive assays for cardiac drug liabilities. We 

have used a four color multi-parameter assay on IN Cell Analyzer to 

profile the phenotypic effects of a large panel of anticancer drugs 

currently in clinical use and in development. Multi-parameter profiling 

and clustering provides an efficient means to characterize cardiotoxic 

drug actions and to gain insights into mechanisms of cell injury. 

Image Analysis Technology Showcase 

1:45-2:15 New Developments in Accelerating Sponsored by 

the High-Content Screening Work Flow 

Grischa Chandy, Product Manager, Cellular Imaging, 

Molecular Devices, LLC 

Evan F. Cromwell, Ph.D., Director, Research, Molecular Devices, LLC 

Next-generation high-content tools for imaging offer automated 

techniques for modeling diseases and predictive toxicology. We 

will present examples of multi-parametric assays for testing the 

effects of compounds in a variety of assays and highlight the new 

innovative software that provides step-by-step assistance for designing 

customized, reusable, and distributable analysis algorithms. 

2:15-2:45 Get the Big Picture in Phenotypic Sponsored by 

Screening for Drug Discovery 

Oliver Leven, Head, Professional Services, Genedata 

High Content Screening is now routine in drug 

discovery, but integration of the HCS dataflow is still problematic. 

HCS images from phenotypic screening are reduced to numerical 

results and then are stored in isolated, instrument-specific image data 

management systems. Often there is no integration with central result 

data warehouses, and assay and compound results may be stored 

without any reference back to original image data. We will present 

a data management workflow that starts after initial image capture, 

supports data analysis and interpretation, and allows result mining using 

past experiments and maintains references to well images. 

2:45-3:15 Analysis and Reporting for High- Sponsored by 

Content Screening with FCS Express 4 Image 

Cytometry by De Novo Software 

David Novo, Ph.D., President & CEO, De Novo Software 

De Novo Software has developed premier analysis and reporting tools 

for research and clinical flow cytometry for a decade. Leveraging this 

experience, we developed an image cytometry package for HCA. A 

flow cytometry analysis environment for image cytometry data allows 

fully interactive data and image review at a single-cell level. Create 

sophisticated reports with 1-click. 

Sponsored by 

3:15-4:15 Refreshment Break in the 

Exhibit Hall with Poster Viewing 

PRESENT A POSTER AND SAVE $50! 

Cambridge Healthtech Institute encourages attendees to gain further 

exposure by presenting their work in the poster sessions. To secure a poster 

board and inclusion in the conference materials, your abstract must be 

submitted, approved and your registration paid in full by November 16, 2012. 

Register online, or by phone, fax or mail. Please indicate that you would 

like to present a poster. Once your registration has been fully processed, 

we will send an email with a unique link and instructions for submitting 

your abstract using our online abstract submission tool. Please see below 

for more information. 

Reasons you should present your research poster at this conference: 

• Your poster will be exposed to our international delegation 

• Receive $50 off your registration 

• Your poster abstract will be published in our conference materials 

• Your research will be seen by leaders from top pharmaceutical, biotech, 

academic and government institutes 

Note: Posters should be portrait orientation, with maximum 

dimensions of 36 inches wide (3 feet) x 48 inches high (4 feet). 




January 8-11, 2013 


Analysis 

Anniversary 

3:15-4:15 Refreshment Break in the Sponsored by 


Novel Biosensor Assays for Screening 

4:15-4:40 Development, Optimization and Validation of an 

HCS Biosensor Assay to Identify Compounds that Disrupt 

AR-TIF2 Protein-Protein Interactions 

Paul A. Johnston, Ph.D., Research Associate Professor, Pharmaceutical 

Sciences, School of Pharmacy, University of Pittsburgh 

High Transcriptional Initiation Factor 2 (TIF2) coactivator expression levels are 

associated with prostate cancer (CaP) recurrence after androgen ablation 

therapy (AAT). We will describe the development and optimization of a novel 

high-content image-based AR-TIF2 protein-protein interaction biosensor 

(PPIB) assay that exploits features of protein targeting to organelles, AR and 

TIF2 functional domains, and fluorescent reporters to generate positional 

biosensors to measure and quantify the interactions between AR and TIF2 

in cells. We will validate the performance of the AR-TIF2 PPI HCS assay by 

screening the LOPAC and NIH Clinical Collection compound libraries in two 

distinct formats; to identify compounds that can either block the formation 

or that can disrupt established AR-TIF2 PPI complexes. 

4:40-5:05 Novel Approaches to High-Content Imaging of 

Insulin Receptor Trafficking, Insulin Signaling and Endosomal 

Calcium Signaling 

James Johnson, Ph.D., Associate Professor, Cellular and Physiological 

Sciences, University of British Columbia 

We have generated insulin receptor reporters that do not impair 

signaling function in cells and will present data on insulin receptor 

trafficking and signaling in pancreatic beta-cells. We have developed the 

first calcium biosensor capable of measuring calcium in the lumen on 

the endosome and will present our analysis of the role for endosomes 

as dynamic calcium buffers. We present multiplexing approaches to 

generating rich data sets reporting on insulin signaling. 

5:05-5:30 High-Content Screening for Small Molecule 

Inhibitors of HIV Nef 

Andreas Vogt, Ph.D., Associate Professor, Drug Discovery Institute, 

University of Pittsburgh 

The HIV-1 accessory protein Nef is essential for high-titer viral replication 

and AIDS progression. The cellular activities of Nef are critically dependent 

on a variety of protein-protein interactions, including formation of Nef 

oligomers. Nef mutations that interfere with oligomerization prevent HIV 

replication in cell culture, suggesting that the Nef oligomerization interface 

is a rational target for Nef-directed anti-HIV therapy. In this talk, I will 

present the development and validation of a high-content, bimolecular 

fluorescence complementation assay for Nef dimerization inhibitors. 

HCS at NCATS 

5:30-5:55 Supporting Drug Discovery and Reposition in 

NCATS Using HCS Technology 

Zhuyin (Julie) Li, Ph.D., Biology Team Leader, Division of Pre-Clinical 

Innovation, National Center for Advancing Translational Sciences, NIH 

The mission of the newly created National Center for Advancing 

Translational Sciences (NCATS) in NIH is to catalyze the generation of 

innovation methods and technologies that will enhance the development, 

testing, and implementation of diagnostics and therapeutics across a wide 

range of human diseases and conditions. This presentation will highlight 

successful applications of HCS in target validation, compound screening, 

MOA study, toxicity investigation and drug repositions in NCATS. 

High-Content Image and Data Analysis (continued) 

4:15-4:40 Using New Cell Dyes and Automated Image 

Processing to Evaluate Cellular Responses to Small Molecules 

David W. Andrews, Ph.D., Director and Senior Scientist, Biological 

Sciences, Sunnybrook Research Institute, Toronto; Professor, 

Biochemistry, University of Toronto 

Here we describe a simple approach to quantify the responses in 

adherent cells to small molecules based on multivariate analysis of 

cells stained with new mix and read dyes. These dyes are non-toxic, 

non-fluorescent in water and available in several emission/excitation 

wavelengths compatible with existing HCA instruments. We compare 

multivariate analysis and clustering with more traditional measures 

of analysis and find that it provides high Z’ sensitivity and specificity 

resulting in improved classification in screening. 

4:40-5:05 A Label-Free Random Cell Motility Assay Based 

on Image Correlation Spectroscopy 

Michael Prummer, Ph.D., Scientist, High-Content Screening, 

F. Hoffmann-La Roche AG 

Cell migration is central to embryonic development, wound healing, 

inflammation, and cancer. Sparse metastatic cells or T-cells show 

isotropic and random motion, which is difficult to characterize 

with classical tools like scratch assays. We use image correlation 

spectroscopy (ICS) to quantify the speed and mode of random cell 

motility without labeling, identification or trajectory reconstruction. ICS 

offers a toolbox to analyze free, directed, hindered, or confined random 

walks. The random motility (RAMOT) assay is validated using THP1 

immune cells, cytoskeleton modulators and Monte-Carlo simulations. 

Combining ICS and HCS, the RAMOT assay opens up new routes in 

label-free image-based drug discovery. 

5:05-5:30 Image Analysis and Modeling of 

Cardiomyocyte Hypertrophy 

Jeffrey Saucerman, Ph.D., Assistant Professor, Biomedical 

Engineering, University of Virginia 

Cardiomyocyte hypertrophy plays a key role in the transition to 

heart failure. We are developing automated microscopy and image 

analyses to quantify the hypertrophy dynamics of individual live 

primary cardiomyocytes. I will present two applications. In the first, 

we characterized the kinetics of myocyte hypertrophy in response to 

transient receptor agonists. In the second example we used automated 

imaging to validate model predictions about the quantitative role of 11 

parallel hypertrophy pathways. 

5:30-5:55 An Evolving View of Cancer: High-Content 

Analysis and Mathematical Modeling to Study Cancer Cell 

Heterogeneity and Resistance 

Arijit Chakravarty, Ph.D., Senior Scientist, Modeling and Simulation, 

DMPK, Millenium Pharmaceuticals 

The changing picture of the landscape of carcinogenesis and tumor 

response to therapy frames cancer as a disease of genomic instability 

and somatic Darwinian evolution. Developing realistic model systems 

and methodologies to study heterogeneity and evolution in populations 

of cancer cells would be the first step in leveraging the emerging 

picture of cancer in oncology drug development. In this presentation I 

will discuss the challenges posed by tumor heterogeneity and evolution, 

and the methods by which high-content analysis techniques, coupled 

with mathematical modeling, allow us to study this process. 

6:00-7:00 Networking Reception in the Exhibit Hall with Poster Viewing Sponsored by 

Phenotypic Drug Discovery 




January 8-11, 2013 


Analysis 

Anniversary 

Thursday, January 10, 2013 

7:30-8:15 am Breakfast Presentation Sponsored by 

Spots to Spheres: Parasites to 3-D Cancer Models for Drug Discovery 

Vicky M. Avery, Ph.D., Professor, Chief Investigator and Head, Discovery Biology (Avery Lab), Eskitis Institute for Cell & 

Molecular Therapies, Griffith University 

The focus of our laboratory is drug discovery and how we can improve the process, or obtain more information from the data we generate. This 

presentation will provide exemplars of the image-based assays utilized to identify new small molecule inhibitors and modulators of parasitic 

diseases and cancer, as well as those used to investigate the mechanisms involved. 

8:15-8:25 Award Presentations 

Cellume Award 

Poster Competition 

GE Image Competition 

Sponsored by 

Sponsored by 


8:30-8:55 The Value of Phenotypic Assays to Drug Discovery 

David C. Swinney, Ph.D., CEO, Institute for Rare and Neglected 

Diseases Drug Discovery (iRND3) 

Drug discovery strategies include target-based molecular approaches and 

phenotypic-based empirical approaches. Our recent analysis revealed the 

phenotypic approach as the more successful strategy for first-in-class 

medicines. We rationalized that this success was influenced by the unbiased 

identification of a molecular mechanism of action (MMOA) that contributed to 

a useful therapeutic index. The value and success of phenotypic approaches 

will be further increased through efforts to bridge the gap between molecular 

approaches and phenotypic approaches such that once an effective MMOA 

has been confirmed with phenotypic assays, molecular approaches can be 

efficiently utilized to optimize leads to medicines. 

8:55-9:20 Modern Phenotypic Drug Discovery Is a Viable 

Pharma Strategy 

Jonathan A. Lee, Ph.D., Senior Research Advisor, Quantitative Biology, 

Eli Lilly & Co. 

The majority of first-in-class NMEs originate from phenotypic screening 

(PS); however, this approach is not commonly used by the pharmaceutical 

industry. We addressed perceived issues with PS by conducting an antiangiogenesis 

MTS. Identification of novel chemical scaffolds that are 

differentiated, structurally and mechanistically, from anti-angiogenic standards 

of care and which modulate molecular targets not previously associated with 

angiogenesis demonstrates that PS interrogates relevant biology regardless 

of target validation status. Modern PS combines the biological complexity 

of physiology-based systems with the high-throughput compound testing 

capacity and operational robustness of biochemical methodologies. 

9:20-9:45 High-Content Phenotypic Screening for Target 

Identification at Pfizer: High-Throughput Flow Cytometry 

and Live-Cell Imaging of Human Primary Cells 

Regis Doyonnas, Ph.D., Senior Principal Scientist, Stem Cell 

Engineering and High-Content Screening Lab, Hit Discovery and Lead 

Profiling, PDM-NCE, Worldwide Research & Development, Pfizer 

9:45-10:45 Coffee Break in the Exhibit Hall Sponsored by 

with Poster Viewing 

3-D Cell-Based Assays 


8:30-8:55 Overcoming the Challenges of Implementing 3-D Cell- 

Based Assays in Automated High-Content Screening Workflows 

Anthony M. Davies, Ph.D., Director, Irish National Center for High- 

Content Screening and Analysis (INCHA) 

To meet the needs of our in-house drug discovery and target identification 

programs, we have developed a novel 3-D cell culture system that has 

been specifically designed for use with HCS/A imaging and automated 

liquid handling. Unlike the 3-D assay systems currently used, our 

technology does not rely upon solid gel matrices, scaffolds, micropatterned 

surfaces or hanging drop assay systems to achieve reproducible 

cancer spheroid growth, hence offering advantages in flexibility and ease 

of use in an HCS/HTS environment. In this presentation we examine 

(i) Some of the most commonly used 3-D assay systems and their 

advantages and disadvantages when used in conjunction with HCA; (ii) 

In the same context we will also present our own 3-D cell suspension 

technology and the latest data derived from its use. 

8:55-9:20 3-D Hepatocyte Culture to Profile Compounds 

Marc Bickle, Ph.D., Head, HT Technology Development Studio, Max 

Planck Institute of Molecular Cell Biology and Genetics 

3-D cell culture increases the physiological relevance of cellular systems. 

In this context we have established a 3-D hepatocyte cell culture system, 

where polarized hepatocytes form bile canaliculi. In this system we 

have established assays to probe secretion, endocytosis, mitochondria, 

peroxisomes, autophagy and lipid droplets in order to profile compound 

action on these various cellular pathways. These assays will help predict 

the toxicity of compounds and establish their mode of action. 

9:20-9:50 Sponsored Presentations 

(Opportunities available. Contact Katelin Fitzgerald at kfitzgerald@ 


9:50-10:45 Coffee Break in the Exhibit Hall Sponsored by 

with Poster Viewing 




January 8-11, 2013 


Analysis 

Anniversary 

Studying the Tumor Microenvironment: 

HCA of 3-D Tumor Spheroids 

10:45-11:10 3-D High-Content Screening for the Identification 

of Compounds that Target Chemo-Resistant Tumor Cells 

Carsten Wenzel, High-Content Analysis, Bayer Healthcare 

Limited predictability of cellular assays based on 2-D cell culture for the 

efficacy of drug candidates in vivo has fostered the search for novel culture 

techniques for drug discovery in oncology. Based on significant differences 

in gene expression between cells cultured in 2-D and 3-D, as well as 

more physiological growth conditions that resemble the situation in tumor 

tissue, spheroid-based 3-D cell culture is being recognized as a potential 

test platform to better predict drug efficacies in vivo. Here, we present our 

current work on evaluating and implementing different methods for the 

automated generation of 3-D spheroids in microtiterplates, visualization 

of selected markers, automated microscopy and image analysis. By this 

approach we identified several compounds that specifically target cells in 

quiescent tumor spheroid core regions. Additionally we could show that 

these hits, combined with cytostatics, show a significant increase in cell 

death in our spheroid model compared to single treatment conditions. 

11:10-11:35 Elucidation of Cell Signaling Using High-Content 

Imaging of Patient-Derived Breast Tumor Spheroids with 

Cancer Stem Cell Characteristics 

Fredika M. Robertson, Ph.D., Professor, Experimental Therapeutics, 

Center for Targeted Therapy, Translational Therapeutics Laboratory, 

University of Texas MD Anderson Cancer Center 

Using tumor spheroids, we have identified specific gene signatures 

and genetic abnormalities that are involved in cancer metastasis which 

we find to be associated with changes in cell signaling pathways that 

can be targeted using small molecule inhibitors. This presentation will 

discuss the use of the combination of high-content live-cell imaging, 

confocal microscopy, flow cytometry, and histological approaches that 

allow imaging of these surrogates of metastasis and the evaluation of 

a patient’s tumor cells for drug responses in real time. 

11:35-12:00 pm 4-D MAME Models for Live-Cell Imaging 

of Interactions between Breast Tumor Cells and Their 

Microenvironment: Adapting for Compound Screening 

Bonnie Sloane, Ph.D., Distinguished Professor and Chair, 

Pharmacology and Karmanos Cancer Institute, Wayne State University 

To define druggable pathways involved in breast cancer progression, our 

laboratory has pioneered the development of techniques for functional 

imaging of protease activity associated with live human breast cells and 

of 3- and 4-D co-culture models that recapitulate breast tumor architecture 

as well as the cellular and non-cellular tumor microenvironment, i.e., 

MAME (mammary architecture and microenvironment engineering) 

models. Use of these techniques and models in concert with various 

types of imaging probes has allowed us to image, quantify and follow 

the dynamics of proteolysis in the tumor microenvironment and to test 

interventions that impact directly or indirectly on proteolytic pathways. 

12:00-12:25 The Multicellular Tumor Spheroid: A Systems 

Biology Approach for High-Content Imaging and Cancer 


Daniel V. LaBarbera, Ph.D., Assistant Professor, Pharmaceutical 

Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, 

University of Colorado 

In vitro 3-D models, particularly multicellular tumor spheroids (MCTS), offer 

a systems biology approach to bridge information from in silico, molecular 

target, and 2- D cell-based screening, with in vivo models to increase the 

predictability of cancer drug discovery. Here we will discuss our current 

research targeting the metastatic phenotype in cancer using MCTS models 

suitable for high-content imaging. 





January 8-11, 2013 


Analysis 

Anniversary 

Phenotypic Drug Discovery (continued) 

10:45-11:10 A Chemical Approach to Controlling Cell Fate 

Sheng Ding, Ph.D., William K. Bowes, Jr. Distinguished Investigator 

and Professor, Pharmaceutical Chemistry, Gladstone Institute of 

Cardiovascular Disease, University of California San Francisco 

Recent advances in stem cell biology may make possible new 

approaches for the treatment of a number of diseases. A better 

understanding of molecular mechanisms that control stem cell fate as 

well as an improved ability to manipulate them are required. Toward 

these goals, we have developed and implemented high-throughput 

cell-based phenotypic screenings of arrayed chemical libraries to 

identify and further characterize small molecules that can control stem 

cell fate in various systems. This talk will provide the latest examples 

of discovery efforts in my lab that have advanced our ability and 

understanding toward controlling stem cell fate, including self-renewal, 

survival, differentiation and reprogramming of pluripotent stem cells. 

11:10-11:35 Screening of Protein Kinase Inhibitors to 

Identify Target and Liability Kinases in Axon Growth 

John L. Bixby, Ph.D., Professor, Pharmacology and Neurological 

Surgery; Vice Provost for Research, University of Miami 

We screened 995 protein kinase inhibitors (PKIs) in primary neurons to 

identify critical kinase targets and “hit” PKIs that promote growth of 

neuronal processes. We analyzed the activity profiles of hit and non-hit 

PKIs against a kinase panel, narrowing the set of informative kinases. 

A machine learning model was trained using these kinases and will be 

used to perform in silico searches for novel hits using experimentally 

acquired or predicted activity profiles. Our novel approach allows us 

to search for hits based on biochemical profiles rather than SAR, and 

addresses the problem of PKI polypharmacology. 

11:35-12:00 pm Zebrafish as a Tool for Rapid, in vivo 

Identification of Complex Behavioral Chemotypes 

Andrew J. Rennekamp, Ph.D., Postdoctoral Researcher, 

Cardiovascular Research Center and Division of Cardiology, 

Massachusetts General Hospital, Harvard Medical School 

Zebrafish larvae are well suited for in vivo, high-content screens and 

yet they are neurocompetent organisms exhibiting complex behavioral 

phenotypes with neurophysiology that is remarkably similar to humans. 

Using zebrafish, we have identified several reproducible behaviors 

amenable to high-content screening. By probing these behavioral 

phenotypes with compounds having known pharmacological targets 

in humans, we are able to identify neurological pathways relevant to 

human diseases. Subsequently, we are able use the zebrafish as a 

tool for the identification of novel drugs which perturb those important 

pathways via previously unknown mechanisms. 

12:00-12:25 Image-Based High-Dimensional Profiling of 

Cell Response to Perturbation 

Auguste Genovesio, Ph.D., Group Leader, Image Analysis, Broad 

Institute of MIT and Harvard 

From each image in a high-content experiment, hundreds of cells 

can be detected and analyzed leading to millions of high-dimensional 

cell profiles for an experiment. These profiles may be a rich source of 

information about the corresponding RNAi or small molecule treatments, 

revealing complex and subtle phenotypes and identifying families of 

treatment conditions. How to maximize the insights gained from these 

huge datasets is currently a challenging question for the community. In 

this talk, we discuss the data analysis of high dimensional datasets of 

this kind and show applications in drug discovery and basic research. 

HCA of Stem Cells 


2:00-2:25 Development of Screening Platforms for 

Modifiers of Cell Fate in Human Pluripotent Stem Cells 

April Pyle, Ph.D., Assistant Professor, Eli & Edythe Broad Center 

of Regenerative Medicine and Stem Cell Research, University of 

California, Los Angeles 

Differentiated cells from human pluripotent stem cells (hPSCs) provide an 

unlimited source of cells for use in regenerative medicine. However, prior 

to establishment of patient specific cells for cell therapy it is important 

to understand the basic regulation of cell fate decisions in hPSCs. One 

caveat is that hPSCs survive poorly upon dissociation, which limits 

genetic manipulation because of poor cloning efficiency of individual 

hPSC, and hampers production of large-scale culture of hPSCs. Improving 

our understanding of pathways that are important for hPSC growth 

could improve our ability to culture, differentiate and derive new hPSCs. 

Our large scale screening work provides a comprehensive evaluation of 

chemical space for modifiers of cell fate in hPSCs. 

2:25-2:50 Patient-Specific Cell-Based Disease Models for 


Anne Bang, Ph.D., Director, Cell-Based Disease Modeling and 

Screening, Sanford-Burnham Medical Research Institute 

Patient-specific primary cells and induced pluripotent stem cells (iPSCs) 

complement traditional cell-based drug discovery assays, allowing testing 

on differentiated features not reflected by immortalized lines. We used 

patient cells to develop a phenotypic assay for muscular dystrophy 

that distinguishes between affected and unaffected siblings, faithfully 

recapitulating key molecular features of the disease. A high-content screen 

of patient cells was conducted with the goals of identifying early treatment 

candidates, and probes to gain a greater understanding of underlying 

cellular defects. We will discuss screening results and development of 

iPSC-based models for testing of drugs on disease relevant cell types. 

2:50-3:15 HCS to Discover Drugs for Heart Failure 

Mark Mercola, Ph.D., Professor and Director, Muscle Development and 

Regeneration Program, Sanford-Burnham Medical Research Institute 

There is an urgent need for therapies that reverse the course of 

ventricular dysfunction in heart failure, which remains a leading cause 

of morbidity and mortality in Western countries. Current therapies 

do not treat the underlying cause—death or damage of heart muscle 

cells combined with increased scarring. Our research is focused on 

developing high-content screening assays and instrumentation to 

discover targets and screen for molecules active in cardiac regeneration 

and cardiomyocyte contractility. The presentation will discuss 

modeling regeneration using stem cells in high-throughput analysis of 

cardiomyocyte physiology, and demonstration of small molecules that 

promote differentiation and RNA molecules that sustain contractility in 

the failing heart. 

3:15-4:15 Refreshment Break in the Sponsored by 






January 8-11, 2013 


Analysis 

Anniversary 

Digital Pathology and Tissue Diagnostics 


2:00-2:25 Digital Pathology: Development, Challenges and 

Strategies Using HCA to Measure in vivo Responses 

Joe Trask, Ph.D., Head, Cellular Imaging Core, The Hamner Institutes 

for Health Sciences 

In this talk I will discuss the similarities between digital pathology and 

high-content analysis and how these disciplines when taken together 

are used to extract in-depth multiparameteric data to better understand 

the biological outcome from in vivo response from tissue sections. 

In a test case scenario of a small animal study, rats were treated 

with a synthetic steroidal antiandrogen drug cyproterone acetate 

(CPA) or 2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD); whole 

liver sections were isolated, processed, and labeled with multiple 

probes to identify cellular damage including apoptosis using activated 

caspase-3, TUNEL, and nuclei with either fluorescent or chromagen 

indicators. I will discuss the limitation of imaging technology to capture 

tissue sections, the challenges and the methods employed to address 

autofluorescence in liver tissue and development of a multiplexed 

bioprobe assay using automated image analysis techniques. 

2:25-2:50 High-Content Analysis of Peripheral Patient 

Tissues for Analyzing Biological Processes and Drug 

Response in Neurodegeneration 

Justin D. Boyd, Ph.D, Scientist, Laboratory of Drug Discovery in 

Neurodegeneration, Harvard NeuroDiscovery Center, Harvard 

Medical School, Brigham and Women’s Hospital 

Neurodegeneration is ranked among the top causes of death in the 

United States. The majority of neurodegenerative diseases have no 

effective therapy. A challenge to discover disease-modifying drugs for 

neurodegeneration is that the diseases manifest in the brain. But what 

if we could use blood to monitor a disease process Processes that 

contribute to the degeneration in the brain have been reported in blood. 

We are developing a platform to monitor a disease process in blood cells 

from neurodegenerative disease patients to 1) characterize different 

forms of the disease as they relate to these processes, 2) to identify 

drugs that elicit a modification of a disease-causing dysfunction and 3) to 

identify patients most likely to respond to specific treatments. 




Sponsored by 

3:20-4:15 Refreshment Break in the 


RNAi Screens 

4:15-4:40 SeedSeq—Off-Target Transcriptome Database 

Karol Kozak, Ph.D., Head, Data Handling Unit and High-Content 

Screening, ETH Zurich 

Owing to a tolerance for mismatches and gaps in base-pairing with 

target transcripts, small RNAs could have up to hundreds of potential 

target sequences in a genome, and some small RNAs in mammalian 

systems have been shown to affect the levels of many messenger 

RNAs (off-targets) besides their intended target transcripts (ontargets). 

The Reference Sequence (RefSeq) collection aims to provide 

a comprehensive, integrated, non-redundant, well-annotated set of 

sequences, including mRNA transcripts. We performed a detailed 

analysis of off-targeted transcripts based on latest RefSeq version. 

We developed SeedSeq, an extended version of the RefSeq database 

including information about off-target transcripts. 

4:40-5:05 A Genome-Wide RNAi Screen Using C. elegans 

Identifies a Drug that Protects against Toxicity Associated 

with Protein Aggregation 

Stephen C. Pak, Ph.D., Assistant Professor, Pediatrics, University of 

Pittsburgh School of Medicine 

The accumulation of misfolded protein aggregates within cells is 

a common cause of tissue injury and degenerative disease (e.g., 

Alzheimer’s, Huntington’s, ALS and alpha-1-antitrypsin (AT) deficiency). 

To better understand the genetic factors that influence pathogenesis 

of protein aggregation disorders, we modeled AT-deficiency in a 

microscopic nematode, C. elegans. Transgenic animals expressing the 

mutant AT protein accumulated large intracellular aggregates, similar to 

those detected in human hepatocytes. Using the ArrayScanVTi imaging 

platform, we developed a high-content assay for the quantification 

of misfolded protein accumulation. Using whole, live animals, we 

performed a genome-wide RNAi screen to identify genetic modifiers 

of AT-deficiency. Moreover, using computational methods, we found 

compounds with known inhibitory activity against the genetic modifiers. 

When tested in human cell models of AT-deficiency, one compound 

significantly reduced accumulation of mutant AT protein aggregates. 

5:05-5:30 High-Content Assays to Assess Cellular 

Stress Responses 

Hakim Djaballah, Ph.D., Director, HTS Core Facility, Molecular Pharmacology 

and Chemistry Program, Memorial Sloan-Kettering Cancer Center 

High-content assays provide the ultimate measure of cellular stress 

responses. We explored this feasibility by investigating responses upon 

exposure to chemicals, or during DNA/RNA transfection using an in-house 

developed panel of indicators/reporters. This has allowed for the first time the 

ability to classify both performance and cytotoxicity of these various agents. 

I will describe our established platform and discuss our results, especially 

regarding the use of transient transfection in chemical and RNAi screening. 

6:00-9:00 Dinner Course* 

Biting into Big Data: Advanced Data Analysis for Phenotypic Screening 

Instructor: Mark A. Collins, Ph.D., CEO, PurpleBio Consulting 

With the continued resurgence of phenotypic drug discovery, driven in part 

by high-content technologies (imaging, flow, etc.) there is now a demand 

for appropriate methods of analysis and mining of this rich multi-parameter 

“big data.” This dinner course aims to lay out in practical terms, the 

design of experiments for PDD campaigns and the biological relevance of 

phenotypic “fingerprints”; analysis and quality control of multi-parameter 

phenotypic data; data-mining of phenotypic fingerprints and large-scale 

meta-analyses of phenotypic data coupled with other information, e.g. 

genome and structure activity relationships (SAR). In addition tools, 

resources and strategies for implementing phenotypic screening data 

analysis will be discussed. 

Drawing on data sets from both published papers and case-studies 

from high-content analysis and flow, the course attendees will gain a 

clear understanding of the practical approaches to maximizing the value 

from multi-parameter data and how to implement them in their own 

organization. This course will be a lively mix of interactive discussion, 

presentations and networking. Attendees will receive a comprehensive 

course book and web resources. 

*Separate registration required 




January 8-11, 2013 


Analysis 

Anniversary 

Friday, January 11, 2013 

Live-Cell Imaging Workshop* 

8:30-8:35 am Chairperson’s Opening Remarks 

8:35-9:00 FLIM HCA from Automated Multiwell Plate Assays 

to Live Disease Models 

Paul French, Ph.D., Professor, Physics; Head, Photonics Group, 

Imperial College London 

This talk will outline advances in our multidimensional fluorescence imaging 

platform to assay biomolecular interactions across the scales from cuvette 

and cell-based assays to live organisms. In particular, I will report on a rapid 

automated FLIM-FRET multiwell plate reader based on time-gated imaging, 

including an assay of HIV Gag protein oligomerisation where we achieve Z’ 

factors > 0.6, and on 3-D FLIM and FRET of live zebrafish and mice using 

time-gated imaging combined with optical tomography. 

9:00-9:25 Localization of Tryptophan and NADH Interactions 

Using Three-Photon FLIM-FRET Microscopy 

Ammasi Periasamy, Ph.D., Professor, Biology; Director, W.M. Keck 

Center for Cellular Imaging (KCCI), University of Virginia 

Recent reports link tryptophan (TRP) metabolic activity to cancer 

development and progression. Increased TRP degradation may also occur in 

early-stage breast and lung cancer. To quantitate the TRP and nicotinamide 

adenine dinucleotide (NADH) interaction, we developed a novel three-photon 

excitation (3PE) fluorescence lifetime imaging and Förster resonance energy 

transfer (FLIM-FRET) microscopy method, able to differentiate tumorigenic 

from non-tumorigenic human live cells. Based on perturbation studies, where 

the addition of glycolytic substrates significantly quenches TRP lifetimes in 

tumorigenic HeLa cells, our results demonstrate the potential use of 3PE- 

FLIM-FRET as a tool for screening in the early stages of cancer. 

9:25-9:50 Monitoring Dynamic Protein Interactions in the 

Living Cell Nucleus 

Richard Day, Ph.D., Professor, Cellular and Integrative Physiology, 

Indiana University School of Medicine 

A critical challenge for public health research is to identify the molecular 

mechanisms that function in the dynamic control of the epigenome. The 

heterochromatin protein 1 alpha (HP1α) coordinates a network of protein 

interactions critical for epigenetic regulation during cellular differentiation, but 

the underlying mechanisms are not well understood. We are using frequency 

domain fluorescence lifetime imaging microscopy (FLIM) and Förster 

resonance energy transfer (FRET) to detect steady-state protein interactions 

involving HP1α. Combined with fluorescence correlation spectroscopy (FCS) to 

characterize the sub-nuclear protein diffusion, these techniques provide novel 

insights into protein network interactions inside the living cell nucleus. 




10:20-11:10 Coffee Break 

11:10-11:35 High-Content Screening with Secreted Peptides 

Produced by an in vitro Co-Culture Strategy 

Peter Antinozzi, Ph.D., Assistant Professor, Biochemistry, Wake Forest 

School of Medicine 

The proposed talk features a unique screening strategy which utilizes inwell 

synthesis of secreted proteins. This method circumvents the often 

difficult process of producing bioactive proteins by utilizing co-cultured 

cells which enable efficient post-translational modifications of secreted 

proteins. Two examples are demonstrated with high-content live-cell 

imaging of proliferation and cell survival events. 

11:35-12:00 pm FUEL for Thought: A Novel Approach to 

Detect Spatial Proximity on Mesoscopic Scales in vitro and 

in vivo Using Luminescence Excitation 

Spencer L. Shorte, Ph.D., Director, Plateforme d’Imagerie Dynamique 

(PFID), Imagopole, Institut Pasteur 

Bioluminescence Resonance Energy Transfer (BRET) improves the 

sensitivity of bioluminescence by red-shifting blue photons, and 

provides a measure of molecular co-localization at distances of up to 

10nm. However, BRET detection methods may overlook long-distance, 

radiating energy excitation-emission effects that are significant in the 

bioluminescent detection regime. Fluorescence by Unbound Excitation 

from Luminescence (FUEL) describes this radiating luminescence effect 

that excites fluorophores by epifluorescence at distances far beyond 

10nm, many microns, or even millimeters away in a manner completely 

distinct from BRET. Further, we show that detection of FUEL per se is 

sufficient to provide a detection of long-distance proximity in, and beyond 

the microscopic range both in vitro and in vivo. By enabling detection of 

mesoscopic proximity between luminescent and fluorescent probes in the 

context of living biological tissues FUEL promises utility as a novel tool for 

high-content analysis in cell and animal models. 

12:00-12:25 Tracking “DNA Repair Centers” in Living 

Mammalian Cells 

Sylvain Costes, Ph.D., Principal Investigator, Cancer and DNA Damage 

Response, Lawrence Berkeley National Laboratory 

Upon DNA damage, nuclear sub-domains are formed in the nucleus. These 

radiation-induced foci (RIF) are characterized by the local recruitment of 

DNA damage sensing proteins such as p53 binding protein (53BP1). We 

recently hypothesized that protein recruitment occurs in specific nuclear 

regions called “repair centers.” By integrating a small X-ray device with a 

microfluidics system on a fluorescent light microscope, we can monitor 

various mammalian cells expressing 53BP1-GFP while being exposed to 

ionizing radiation. Using novel RIF counting algorithms and cell tracking 

algorithms, the formation of DNA repair factories will be discussed as a 

function of cell cycle, cell lineage and cell type. 

12:25-12:50 Dynamic Imaging for High-Content Analysis of 

Three-Dimensional Tissue 

David Nolte, Ph.D., Professor, Physics, Purdue University 

An innovative label-free non-invasive imaging technology called Tissue 

Dynamic Imaging (TDI) extracts high content from live three-dimensional 

tissue culture responding to pharmaceuticals. Coherence-gated dynamic light 

scattering captures cellular dynamics through ultra-low-frequency speckle 

fluctuations that encode a broad range of cellular and subcellular motions as a 

new form of functional imaging contrast. The motions are altered by different 

mechanisms of action and generate drug-response spectrograms that act 

as fingerprints for phenotypic profiling. Applications span from early drug 

candidate screening to point-of-care and personalized medicine. 

12:50-1:15 Time-Lapse Imaging of Promoter Reporters in 

Stem Cell Colonies 

Anne L. Plant, Ph.D., Leader, Cell Systems Science Group, Biochemical 

Science Division, National Institute of Standards and Technology 

Much of our knowledge of stem cell lineage progression comes from studies 

involving fixed time points. We are developing image collection and analysis 

protocols for tracking pluripotent and differentiated cells by time lapse 

microscopy. Because the same individual cells and colonies can be observed 

over long times (>4 days), we can examine gene expression dynamics 

and heterogeneity and relate these features with changes in morphology, 

proliferation, location, and differentiation status in individual cells and colonies. 

Temporal features of promoter reporters can provide information that can 

lead to mathematical prediction of the kinetics of cell fate. 

1:15 Close of Conference *Separate registration required

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