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   The ECIS Connection - September 2010           
Links to Articles In This Issue:
Company News - 2010 ECIS Users Meeting
Software Update
ECIS Webinar Schedule
New Publications
Tradeshows 2010
Tip of the Month
ECIS Cartoons
Company News

2010 ECIS Users Meeting

ECIS 2010 Group Photo

The ECIS Users meeting, held August 24-27 at the historic Rensselaerville Institute in Upstate NY, was a great success both scientifically and socially. Over 40 attendees from the United States and Europe met for three days of talks, poster sessions and workshops. Research topics varied, but all had the common theme of employing ECIS measurements to discern changes in cell behavior. It is difficult to single out any particular talk, but one that stimulated much interest was the first description of the use of ECIS to follow adipose-derived stem cell differentiation.
 
On Wednesday evening, Ivar Giaever, CTO of Applied BioPhysics and recipient of the 1973 Nobel Prize in physics, gave a stimulating and humorous talk entitled "How to Win a Nobel Prize." The Rensselaerville Institute staff provided delicious meals throughout the meeting including a wonderful Thursday night barbecue held outdoors under starry skies. In addition, each night there were fireside discussion sessions where ECIS users exchanged ideas, shared common experiences and formed new friendships.
 
We look forward to next year where tentative plans are in place for another ECIS Users Meeting in Regensburg, Germany in late July 2011. Stay tuned for details.
 
To view pictures of the meeting please visit our Facebook Page.

New ECIS Arrays

8WCP (Cell Proliferation)  -- Each of the 8 wells has two sets of 7.5mm x 0.5mm rectangular active electrodes located on inter-digitated fingers to provide measurements of cells.  Each well has a substrate area of 0.8 cm^2 and a maximum volume of 600 μL. On average, with a confluent layer, approximately 4000 to 8000 cells will be measured by the electrodes.

The 8WCP arrays are designed to monitor larger numbers of cells, sampling over the entire bottom of the well. Because of the relatively high number of cells, impedance fluctuations due to micromotion are smoothed out and do not obscure subtle changes in impedance due to the experimental conditions.

Recommended for use with Cell Proliferation Assays.

Line (Wounding) -- Each of the 8 wells contains a single linear electrode with dimensions of 667µm x 150µm and an area equal to that of our standard 250µm circular electrodes. Each well has a substrate area of 0.8 cm2 and a maximum volume of 600μL. On average, with a confluent cell layer, approximately 50 to 100 cells will be measured by the electrode, but even a single cell can be observed.

Recommended for use with cell migration measurements via automated wound healing.

2W4x10E -- This array configuration was developed in collaboration with Yang Wang of the University of Chicago. Dr. Wang's requirements were for arrays with a large surface area to volume ratio to allow for rapid gas exchange within the media. These arrays have greatly aided Dr. Wang in his ECIS anoxia experiments. Each of the 2 circular 25 mm diameter wells contain four independent sets of ten 250 μm diameter active electrodes.  In addition, the center of these arrays are left clear, without gold or photo-resist allowing for live cell Fluorescent microscopy with a standard inverted microscope.

8F10E -- This is a specialized array with 8 sets of 10 active 250 μm diameter electrodes located in the central region at the base of a flow channel measuring 50 mm long, 5 mm in width and 0.4 mm in height.

The flow array is useful for ECIS measurements of cells under perfused conditions or to mimic the shear stress endothelial cells experience in vivo. The channel has an area of 2.5 cm2 and a channel volume of 100 μL. On average, a confluent layer of approximately 500 to 1000 cells will be monitored by each set of electrodes.

HNA Arrays --  We can customize any of our 8 well arrays with a thin substrate, the thickness of a coverslip. Recommended for use with High Numerical Aperture Microscopy.


ABP Beta Tests a New Wounding Method: "The Electric Fence"

The Electric Fence is an experimental alternative method to the conventional ECIS wound healing protocol. Instead of growing cells to confluence and then wounding the cells with high current, high current is used to prevent cells from growing on the electrode surface. Cells grow on all areas of the well bottom except the "fenced" electrode.
 
Once a confluent layer has been established around the electrode, the "electric fence" is turned off, and cells migrate inward to fill the open area. This population of the electrode is monitored with normal ECIS measurements, and, as in the wounding assay, cells must move the radius of the electrode to bring the final impedance values up to those of the control cell-covered electrodes.

This alternative method is particularly useful in cases where standard ECIS wounding does not work well because cells do not easily release after wounding. ABP is currently beta testing the Electric Fence with select customers. It uses the current ECIS wounding hardware and requires only a change to the software.  If you are also interested in acting as a beta tester, please contact Christian Renken at renken@biophysics.com.  


New Products

CO2 Microscope Stage Incubator System

stage incubatorThe Model S1S ECIS stage incubator system is a turn-key system to provide proper temperature and atmospheric conditions to allow simultaneous ECIS and optical measurements. The device fits on the stage of an inverted tissue culture microscope and accommodates up to two 8 well ECIS arrays. Temperature, monitored by a thermocouple located in the chamber, is controlled by a circulating water bath connected to the device. Temperature output can be logged with the ECIS data. Two gas flow rotameters and a bubble humidifier are included to control the atmosphere within the incubator. The system includes all necessary tubing and fittings.


For more information visit: Stage Incubator


Hypoxia Chamber

hypoxia chamber
The ECIS Environmental Control Cabinet allows researchers to monitor cell behavior and conduct ECIS experiments in a controlled gas environment.The cabinet will accept either the 16 or 96 well ECIS Station and fits within a standard tissue culture incubator that is used to regulate the cabinet's temperature. Optional gas sensors within the cabinet are used to measure and regulate the concentration of oxygen and/or carbon dioxide within the cabinet. Pull-out shelves provide easy access to the array holder.
 

Cabinet size: 8.25" H x 16" W x 14" D
Overall footprint: 13" H x 16" W x 15" D
 
For more information visit: Hypoxia Chamber


ECIS Early Career Grant

Applied BioPhysics would like to help young scientists obtain funding. The ECIS mini-grant is aimed at early career scientists who are applying for their first RO1 grant. For a researcher wanting to use ECIS technology to achieve their research goals, Applied BioPhysics will provide an ECIS instrument, ECIS arrays, and consultation in order to generate preliminary data to support the applicants RO1 proposal. Interested scientists should submit their research plan with a cover letter explaining how ECIS technology can be used to achieve their specific objectives. Applied BioPhysics will evaluate proposals based on scientific merit, suitability with ECIS technology and novelty. 


To apply please send a resume, RO1 research plan and cover letter to Christian Renken at renken@biophysics.com.



Software
 
The latest version of ECIS software is v1.2.55 (for the PC). 

Major Changes:

1. When using the software you may notice a black console window in the background. This is normal and the window must not be closed, but can be minimized. Any messages from the software will appear in the console window and can be reported to ABP in case of problems.
 
2. When acquiring data the GUI (graphical user interface) will be partially locked. This can be overridden by un-checking the 'lock' icon on the toolbar. The GUI lock will disable all menu items, the ability to change datasets or switch to Analysis. Only certain toolbar buttons will be available; zoom, pan, data cursor, legend, full screen and thumbnails are all disabled.
 
To minimize potential Windows issues during acquisition the following steps are recommended:
  • Turn off Wifi and disconnect from the internet
  • Disable any Bluetooth devices and stop any Bluetooth services or taskbar applications
  • Disable the webcam and any Bluetooth devices from the BIOS
  • Close all other applications
  • Make sure any system updates have been installed so Windows doesn't force an update
  • Disable Automatic Updates
  • Uninstall any unnecessary software for applications that appear on the taskbar
  • Ensure at least 1GB of disk space is available
  • Start a fresh copy of the ECIS Software prior to starting acquisition
  • Ensure all USB cables are firmly attached
  • Check the Power Management profile is 'Always On'
A number of improvements are the result of suggestions received from the recent ECIS users meeting, including:
  • Improved modeling interface
  • Export of Error Bar data when exporting Graph Data (File | Export | Graph Data)
  • Different color palettes for 8 well arrays (Edit | Color Palette)

Download and install the update from:

 
If your system is on the internet, go to 'Help | Check for Updates' in the software.
 
You can also install ECIS software on different computers for offline analysis using the CD that came with your system.
 
You can also download the base files from:
 
Then install the latest update. Contact info@biophysics.com for the unzip password.

ECIS Webinar Schedule 2010

ECIS application webinars review the topics listed below in 20 minute, web-based, interactive seminars presented by Applied BioPhysics president and co-founder, Dr. Charles Keese.

All webinars are held at 11:00am EST. To register for a webinar, please go to:
https://appliedbiophysics.webex.com
  • Automated Cell Migration - September 21, 2010
  • Barrier Function Assays - October 5, 2010
  • Real-time Electroporation and Monitoring - October 19, 2010
  • Cell Attachment and Spreading Measurements - November 2, 2010
  • Signal Transduction Assays - November 16, 2010
  • Toxicology with ECIS - December 7, 2010
For a more detailed description of each webinar, please visit: http://www.appliedbiophysics.com/contactUs/webinar.html.
 
New Publications

Analysis of Matrix-Dependent Cell Migration with a Barrier Migration Assay
Sven Kroening and Margarete Goppelt-Struebe. Sci. Signal. 2010; 3:pl1. 
 
Rac1 Recruits the Adapter Protein CMS/CD2AP to Cell-Cell Contacts. Trynette J. van Duijn, Eloise C. Anthony, Paul J. Hensbergen, André M. Deelder, and Peter L. Hordijk. J. Biol. Chem. 2010; 285:20137-20146. 
 
ADAM15 regulates endothelial permeability and neutrophil migration via Src/ERK1/2 signaling. Chongxiu Sun, Mack H. Wu, Mingzhang Guo, Mark L. Day, Eugene S. Lee, and Sarah Y. Yuan. Cardiovasc Res. 2010; 87:348-355. 
 
Netrin-4 induces lymphangiogenesis in vivo. Frederic Larrieu-Lahargue, Alana L. Welm, Kirk R. Thomas, and Dean Y. Li. Blood. 2010; 115:5418-5426. 
 
Calcium/Calmodulin-dependent Protein Kinase II Delta 6 (CaMKII6) and RhoA Involvement in Thrombin-induced Endothelial Barrier Dysfunction. Zhen Wang, Roman Ginnan, Iskandar F. Abdullaev, Mohamed Trebak, Peter A. Vincent, and Harold A. Singer. J. Biol. Chem. 2010; 285:21303-21312. 
 
FoxM1 regulates re-annealing of endothelial adherens junctions through transcriptional control of β-catenin expression. Muhammad K. Mirza, Ying Sun, Yidan D. Zhao, Hari-Hara S.K. Potula, Randall S. Frey, Steven M. Vogel, Asrar B. Malik, and You-Yang Zhao. J. Exp. Med. published 26 July 2010, 10.1084/jem.20091857. 
 
Effects of negative pressures on epithelial tight junctions and migration in wound healing. Chih-Chin Hsu, Wen-Chung Tsai, Carl Pai-Chu Chen, Yun-Mei Lu, and Jong-Shyan Wang. Am J Physiol Cell Physiol. 2010; 299:C528-C534. 
 
Adipose Stem Cell Treatment in Mice Attenuates Lung and Systemic Injury Induced by Cigarette Smoking. Kelly Schweitzer, Brian H Johnstone, Jana Garrison, Natalia Rush, Scott Cooper, Dmitry O Traktuev, Dongni Feng, Jeremy J Adamowicz, Mary Van Demark, Amanda J Fisher, Krzysztof Kamocki, Mary Beth Brown, Robert G Presson, Jr, Hal E Broxmeyer, Keith L March, and Irina Petrache. Am. J. Respir. Crit. Care Med. published 13 August 2010, 10.1164/rccm.201001-0126OC 
 
An efficient analysis of nanomaterial cytotoxicity based on bioimpedance. K Kandasamy, CS Choi, and S Kim. Nanotechnology. 2010; 21: 375501. 
 
miR-802 regulates human angiotensin II type 1 receptor expression in intestinal epithelial C2BBe1 cells. Sarah E. Sansom, Gerard J. Nuovo, Mickey M. Martin, Sainath R. Kotha, Narasimham L. Parinandi, and Terry S. Elton. Am J Physiol Gastrointest Liver Physiol. 2010; 299:G632-G642. 
 
Tumour suppressor function of MDA-7 / IL-24 in human breast cancer. N Patani, A Douglas-Jones, R Mansel, W Jiang, and K Mokbel. Cancer Cell Int. 2010; 10:29. 

 
_______________________________________________________________________

Have you recently published an article that includes the use of ECIS?
If so, submit your publications to Applied BioPhysics via email to Nancy Vlahos at vlahos@biophysics.com. We will announce your article in our newsletter, post it on our website and send you 2 FREE 8 well arrays!

 
Visit Us at Upcoming Events

Applied BioPhysics will have ECIS demonstrations and informational literature at the following tradeshows and events: 

American Society for Cell Biology
ASCB 50th Annual Meeting
December 11-15, 2010
Philadelphia, PA

Tip of the Month:

Improving Experiment Repeatability with Cysteine

Either electrical stabilization (see June 09 Newsletter) or cysteine treatment is strongly recommended to enhance experimental repeatability and to minimize variation between ECIS wells. The electrical stabilization method is very convenient as it is built into the ECIS software and stabilization is actuated with a simple click of the mouse.  That being said, it is necessary to allow 30 minutes or longer after electrical stabilization before the electrodes will settle into their final equilibration values. 
 
The use of cysteine can accomplish the stabilization of the electrodes quickly, with relatively little effort and does not require the equilibration period following treatment. To accomplish this, the well should be flooded with a 10 mM sterile solution of cysteine in water.* The cysteine will form a covalent sulfur-gold linkage with the electrode surface displacing small molecules that have adsorbed to gold surface over time and stabilizing the impedance. After exposure (5 minutes or more), the cysteine solution can be rinsed out of the well, proteins adsorbed if desired, and the wells inoculated with cells. This cysteine layer provides a hydrophilic substrate that is excellent for protein adsorption and ultimately cell attachment and spreading. 
 
For more information on improving experiment repeatability, please visit the ECIS FAQ page of our website.

*Available from Applied BioPhysics
 


ECIS Humor

Need a good laugh? Visit the ECIS Cartoons page of our website to view cartoons by Catherine, our in-house cartoonist, to start your day with a smile.

Are you the creative type? Submit one of your own cartoons; if we post it on our website we will send you a free array!

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