| The ECIS Connection - July 2012 |
ECIS Training Schedule
Sept 20 & 21 2012
Applied BioPhysics is offering a two day in depth ECIS training course. The course will include both lecture and wet lab training. The training will take place at the Applied BioPhysics facility in Troy, NY. Training will be conducted by Dr. Charles Keese, Dr. Christian Renken and Catherine Toniatti.
Attendance will be limited to a maximum of 6 people.
Cost is $950 per person. This includes two nights of lodging, transportation to and from the hotel to the Applied BioPhysics facility, lunch on both days and dinner Thursday evening. To register, contact Wendy Ladouceur for a registration form at 518-880-6860, firstname.lastname@example.org or download the form from our website, biophysics.com.
ECIS-related topics to be covered include:
- Experimental design and array selection
- Array preparation and stabilization
- Obtaining good well-to-well and experiment-to-experiment repeatability
- Applying extracellular matrix proteins to the electrodes
- Techniques for array inoculation
- Techniques for addition of compounds to ECIS wells
- Basics of the ECIS software for data acquisition
- Advanced features of the ECIS software for data acquisition
- Basics of the data analysis software
- Advanced features of the data analysis software
- Basics of impedance measurements
- Theory behind ECIS
- Simple and complex impedance and the value of R and C in cell measurements
- Selecting the AC frequency or frequencies for experiments
- Modeling ECIS data
- In situ electroporation
- Cell migration measurements with the ECIS "wound-healing" assay
- Cell migration measurements with the "electric fence"
- A survey of cell biology applications using ECIS
- Fluorescence staining of cells on the ECIS array
The following is the link to the handbook: http://biophysics.com/PDFS.php
ECIS Handbook Includes:
Choosing the right array
Coating the electrodes with proteins and other macromolecules
Program of Choice
The latest Windows version of the ECIS software is v1.2.98: To download click on the following link:
You can find which version you are running by looking at the splash screen on start-up, or by selecting Help | About.
This version has the ability to modify the legend text that is displayed on the graph plot.
By default the legend will be the well ID (e.g. A1, A2, A3 etc). The Legend can be turned on from the toolbar button. The Legend can then be edited by selecting Edit | Legend (for 16 wells or less). If you have trouble getting the legend to display, first turn on the legend, then export the plot to a separate window by selecting Edit | Export Graph.
Due to a limitation in the Windows timing code, the maximum time for an ECIS experiment was 596 hours (24 days). This limitation has now been overcome in version v1.2.98, and there is now no theoretical limit for ECIS experiments (only how long the cells can be kept alive). It is also not recommended to change the system time during an experiment.
Exporting ECIS Graphs to Word or PowerPoint
You can export a displayed graph to another program such as Word or PowerPoint in several ways:
- From the Edit menu select Copy Graph. Switch to the other program and Paste. The pasted graph can then be rescaled to fit the page or slide.
- From the Edit menu select Export Graph, or click the Export button under Display Options in the bottom right-hand of the GUI. This will open a separate window with dimensions specified in Display Options (default 800 x 600 pixels). From the File menu select Save As... and then a Save As Type that is compatible with the other program, e.g. TIF or JPG. The output bitmap will be the same size as the window.
- Use the options under Edit | Plot Style to change the line style and width. The line colors can be changed from Edit | Color Palette. If you want a specific color the Edit | Color Palette | Manual option will allow a color to be defined as Red, Green and Blue values that range from 0to 1.
If your computer is connected to the internet, you can update to the latest ECIS software by selecting Help | Check for Updates. Otherwise, you can manually download the update by pasting the following url into your browser: http://www.biophysics.com/software/ECIS_Software_Update.msi
New System Install
If you wish to install the ECIS software on a separate computer for off-line analysis, you first need to download the following file (180Mb), unzip it and run 'Install.bat': by pasting the following url into your browser: http://www.biophysics.com/software/ECIS_Web_Install.zip
The unzip password is 'NewECISSoftware'. You should then update your ECIS software to the latest version as described above.
USB Driver Install
If you need to install the software on a new system for acquiring data, follow the instructions above and then install the USB driver. The latest Edgeport driver is v5.30 and is included with with the software. It can be installed by selecting Help | Edgeport USB | Install Driver, then extract the files to 'C:\EdgePort Drivers'. After plugging in the USB cable, use the 'New Hardware Wizard' to manually select the driver.
Windows 7 users in particular will need to use the Edgeport software to verify that the COM ports are numbered sequentially. Select Help | Edgeport USB | Configuration Utility.
If you are using a Mac system please contact us for upgrade instructions.
Please note that the manual has recently been updated and is available as a PDF from Help | Manual.
Default Experiment Type
If you typically collect data with multiple frequencies, you can set this as the default experiment type by selecting Default Expt. Type from the Acquire menu, and then select MFT. The software will then start with that mode selected. You can also choose SFT or RTC if desired.
Set Scan Frequencies
You can manually specify the frequencies to be applied during a MFT using the Acquire | Set Scan Frequencies | Manual option. Frequencies are normally separated by a factor of 2 or 4 for analysis on a logarithmic scale. There is also a predefined option for '4/16/64 kHz' which gives a quick 3 frequency scan for cell monitoring, but is not suitable for modeling.
|ECIS Webinar Schedule 2012
ECIS application webinars review the topics listed below in 20 to 30 minute, web-based, interactive seminars presented by Applied BioPhysics president and co-founder, Dr. Charles Keese. https://appliedbiophysics.webex.com and scroll to the webinar date of interest.
All webinars are held at 11:00am EST. To register for a webinar, please go to:
Signal Transduction Assays - 11:00 AM EST
September 4, 2012
Toxicology with ECIS - 11:00 AM EST
September 18, 2012
ECIS Theory - 11:00 AM EST
October 9, 2012
Cell Invasion / Extravasation Assays - 11:00 AM EST
October 23, 2012
Automated Cell Migration - 11:00 AM EST
November 13, 2012
Barrier Function Assays - 11:00 AM EST
November 27, 2012
Real-time Electroporation and Monitoring - 11:00 AM EST
December 11, 2012
For a more detailed description of each webinar, please visit: http://www.biophysics.com/webinar.php
Serine/Threonine Phosphatase Stp1 Mediates Post-transcriptional Regulation of Hemolysin, Autolysis, and Virulence of Group B Streptococcus Kellie Burnside, Annalisa Lembo, Maria Isabel Harrell, Michael Gurney, Liang Xue, Nguyen-Thao BinhTran, James E. Connelly, Kelsea A. Jewell, Byron Z. Schmidt, Melissa de los Reyes, Weiguo Andy Tao, Kelly S. Doran, and Lakshmi Rajagopal (2011) J. Biol. Chem. 286:44197-44210
The Hedgehog Pathway Promotes Blood-Brain Barrier Integrity and CNS Immune Quiescence
Alvarez, J. I., Dodelet-Devillers, A., Kebir, H., Ifergan, I., Fabre, P. J., Terouz, S., Sabbagh, M., et al. (2011).
Science (New York, N.Y.), 1727. doi:10.1126/science.1206936
Disruption of Nrf2 Signaling Impairs Angiogenic Capacity of Endothelial Cells: Implications for Microvascular Aging M. Noa Valcarcel-Ares, Tripti Gautam, Junie P. Warrington, Lora Bailey-Downs, Danuta Sosnowska, Rafael de Cabo, Gyorgy Losonczy, William E. Sonntag, Zoltan Ungvari, and Anna Csiszar (2012). J Gerontol A Biol Sci Med Sci., 10.1093/gerona/glr229
ROCK and cAMP Promote Lymphatic Endothelial Cell Barrier Integrity and Modulate Histamine and Thrombin-Induced Barrier Dysfunction Jerome W. Breslin. Lymphatic Research and Biology. March 2011,
9 (1):3-11. doi:10.1089/lrb.2010.0016
Endothelial E-Type Prostanoid 4 Receptors Promote Barrier Function and Inhibit Neutrophil Trafficking Konya, V., Kampitsch, N. & Theiler, A. Journal of Allergy and Clinical Immunology (2012).doi:10.1016/j.jaci.2012.05.008
Measurement of Cellular Chemotaxis with ECIS/Taxis KM Pietrosimone, X Yin, DA Knecht, and MA Lynes
J Vis Exp. 2012
Long-term storage and impedance-based water toxicity testing capabilities of fluidic biochips seeded with RTgill-W1 cells LM Brennan, MW Widder, LE Lee, and WH van der Schalie Toxicol In
Vitro. 26: 736-45 (2012)
Ethanol Disrupts Vascular Endothelial Barrier: Implication in Cancer Metastasis Xu M, Chen G, Fu W, Liao M, Frank JA, Bower KA, Fang S, Zhang Z, Shi X, Luo J. Toxicol Sci. 2012 May;127(1):42-53. Epub 2012 Feb 13
IQGAP1 is necessary for pulmonary vascular barrier protection in murine acute lung injury and pneumonia M. Bhattacharya, G. Su, X. Su, J. A. Oses-Prieto, J. T. Li, X. Huang, H. Hernandez, A. Atakilit, A. L.
Burlingame, M. A. Matthay, and D. Sheppard Am J Physiol Lung Cell Mol Physiol. 2012; 303:L12-L19
Agonist of growth hormone-releasing hormone reduces pneumolysin-induced pulmonary permeability edema (Feb 24) Rudolf Lucas, Supriya Sridhar, Ferenc G. Rick, Boris Gorshkov, Nagavedi S. Umapathy, Guang Yang, Aluya Oseghale, Alexander D. Verin, Trinad Chakraborty, Michael A. Matthay, Evgeny A. Zemskov, Richard White, Norman L. Block, and Andrew V. Schally PNAS. 2012; 109:2084-2089
ISG15 disrupts cytoskeletal architecture and promotes motility in human breast cancer cells
Shyamal D Desai, Ryan E Reed, Julian Burks, Laurence M Wood, Ashok K Pullikuth, Arthur L Haas, Leroy F Liu, Jerome W Breslin, Sally Meiners, and Surendran Sankar Exp Biol Med. 2012; 237:38-49
ERG controls endothelial cell permeability via transcriptional regulation of claudin-5 (CLDN5)
Lei Yuan, Alexandra Le Bras, Anastasia Sacharidou, Kiyoshi Itagaki, Yumei Zhan, Maiko Kondo, Christopher V.
Carman, George E. Davis, William C. Aird, and Peter Oettgen J. Biol. Chem. published 10 January 2012, 10.1074/jbc.M111.300236
ADAM10 Mediates Vascular Injury Induced by Staphylococcus aureus α-Hemolysin Michael E. Powers, Hwan Keun Kim, Yang Wang, and Juliane Bubeck Wardenburg The Journal of Infectious Disease. published 2 April 2012, 10.1093/infdis/jis192
Atrial natriuretic peptide enhances microvascular albumin permeability by the caveolae-mediated transcellular pathway (Feb 24) Wen Chen, Birgit Gaßner, Sebastian Börner, Viacheslav O. Nikolaev, Nicolas Schlegel, Jens Waschke, Nadine Steinbronn, Ruth Strasser, and Michaela Kuhn Cardiovasc Res. published 29 November 2011, 10.1093/cvr/cvr279
Angiotensin II increases the permeability and PV-1 expression of
endothelial cells Csaba Bodor, János Péter Nagy, Borbála Végh, Adrienn Németh, Attila Jenei,
Shahrokh MirzaHosseini, Attila Sebe, and László Rosivall Am J Physiol Cell Physiol. 2012; 302:C267-C276
Have you recently published an article that includes the use of ECIS? If so, submit your publications to Applied BioPhysics via email to Dr. Christian Renken at email@example.com. We will announce your article in our newsletter, post it on our website and send you 2 FREE 8 well arrays!
Representatives from Applied BioPhysics will be at the following tradeshows and events:
Stony Brook University
August 8, 2012
Stony Brook, NY
American Society for Cell Biology
December 16 - 18, 2012
San Francisco, CA
|Tip of the Month:
Cell Behavior Upon Complete Removal Of Medium In A Well
As many ECIS users have discovered, impedance measurements record cell behaviors that are subtle and often cannot be discerned with microscopic observations. One of these behaviors is the response of cells upon exposure to an air-water interface. Commonly in the process of changing medium or adding experimental compounds, one completely aspirates the medium over the cell monolayer exposing the cells to a thin layer of liquid at the air interface. If one carries out this manipulation and then follows the response of the cells using ECIS, it is clear this procedure can elicit substantial impedance changes requiring several minutes before equilibrium is achieved. This response of the cells is normal but it can sometimes obscure impedance changes that one wishes to detect - e.g. in the addition of a compound to activate a receptor. For this reason, we suggest that drug addition be carried out without complete removal of medium from ECIS wells. See the ECIS Newsletter Tip for Sept 2009
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!