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   The ECIS Connection - October 2013              

 

 

ECIS School Schedule

 

January 30 - 31, 2014

 

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. Ivar Giaever, Dr. Christian Renken, Dr. Judith Stolwijk and Catherine Toniatti-Yanulavich.

 

Attendance will be limited to a maximum of 6 people.

 

Cost is $1,250 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, ladouceur@biophysics.com 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 

 

 

 

Dr. Ivar Giaever Travels to India  

 

 

 

  

 

 

 

 

 

 

 

 

 

 

 

 

40 Years Later

 

Niskayuna's Ivar Giaever recalls the morning he got word of winning the Nobel Prize

 

October 23, 1973 was a big day at the General Electric Research Laboratory in Niskayuna. Everyone and everything was ready to help celebrate the 100th birthday of physicist William D. Coolidge, the man who vastly improved the efficiency of X-ray machines and light bulbs. Coolidge was still very much alive and would be there at the festivities that day with his colleagues, sipping champagne and eating a large cake adorned with 100 candles made to look like tungsten lamps. What could possibly top that?

 

Well, at 6 a.m. that morning, GE supervisor Milan Fiske got a phone call from a friend in Stockholm, Sweden, with some great news: The Nobel Prizes had just been announced and Ivar Giaever, a native of Norway and a GE employee for the previous two decades, had just been named the Nobel winner in physics.

 

The Coolidge celebration had just been trumped.

 

"They had brought in all this champagne and were going to celebrate Coolidge's birthday, but they ended up using the champagne on me," remembered Giaever, who became just the second GE scientist to win such a prize, joining the 1932 Nobel recipient for chemistry, Irving Langmuir. "It was a surprise to me. I had no idea."   

 

 

An unwanted limousine

 

Fiske had called Giaever after getting off the phone with Stockholm, and gave him the news. Around 8 that morning, a limousine appeared in Giaever's driveway on Van Antwerp Road, less than half a mile away from the research lab. Giaever sent it away.

 

"I bought the house because it was close to work, and typically I walked to work," said Giaever. "When the limousine came I said, 'No, I don't want to go.' About a half hour later the guy drove back and told me I had to take the limousine. So I went and they had a big press conference that morning. It was all amazing. I thought at first it was a joke, but I knew Milan wasn't the kind of person to joke that way. After he called me, the phone didn't stop ringing."

 

The phone continued to ring all day at work, and fortunately for Giaever, co-worker Howard Hart helped him deal with the influx of calls.

 

"It was great excitement and great fun," said Hart, who retired in 1995. "I was picking up his phone and answering all kinds of questions. 'Yes,' I would tell them. 'He is from Norway but he is a citizen of the United States.' So much was going on I can't even remember how Ivar took it. He doesn't have a very expressive face, but I knew he had done some nice work and that he was very proud. We all appreciated his success."

 

George Wise had started at the research lab just a few months earlier.

 

"We all got to go down to the lobby and have a glass of champagne," said Wise, "I was a new employee and I thought, 'Hey, this is a great place to work.' I didn't meet Ivar until after he won the prize, but it was one day I'll always remember, and the universal comment made that day was, 'It couldn't have happened to a nicer guy.' "

 

 

 

A 44-year-old laureate

 

Giaever was 44 when he won the Nobel, a prize he shared with Dr. Leo Esaki of IBM and Dr. Brian D. Josephson of the University of Cambridge in England. According to a press release by The Nobel Foundation, Giaever and Esaki were awarded the prize "for their experimental discoveries regarding tunneling phenomena in semiconductors and super conductors, respectively." Josephson earned his recognition "for his theoretical predictions of the properties of a supercurrent through a tunnel barrier, in particular those phenomena which are generally known as the Josephson effects."

 

Giaever, his wife Inger and all four of their young children flew to Stockholm for the awards ceremony on Dec. 10, 1973.

 

"The Nobel Foundation paid for me, and GE was nice enough to pay for my wife and the kids," said Giaever. "We stayed at a very nice hotel in Stockholm. It was a very nice time."

 

Giaever had been experimenting with tunneling throughout the 1960s, contributing greatly to the study of phenomena that occur at temperatures near absolute zero. He had left Norway in 1952 after graduating from the Norwegian Institute of Technology and landed in Canada where he began working as a mechanical engineer at the General Electric plant in Petersburgh, Ontario. Never a good student, Giaever finally found out what he wanted to do with his life when he transferred to the GE plant in Schenectady in 1954.

 

"I was very lazy in college and really didn't learn that much," said Giaever. "When I came to Schenectady and the research lab, I saw people writing on blackboards, talking to each other, walking back and forth. They were doing research. I was shocked. I said to myself, 'This is what I want to do,' but I was 30 years old by the time I found that out. I never knew you could get paid for doing research."

 

Doctorate at RPI

 

While he was working at GE, Giaever began taking classes at Rensselaer Polytechnic Institute in Troy and eventually approached his boss, Roland Schmitt, about becoming a physicist.

 

"I figured even if he flunked out of physics, he's still a good applied mathematician," Schmitt told the Gazette in 1998, speaking of Giaever. He didn't flunk. In 1964, the year he became a U.S. citizen, he also earned a doctorate from RPI in theoretical physics.

 

"My father and mother were very happy to see me get my doctorate," remembered Giaever. "My mother had always said that I should be a locksmith because I could always open all the doors. And I had thought I wanted to be an engineer. I had no idea that I could be a scientist and get paid for it."

 

Charlie Bean, John Fisher, Bill Johnson and Walter Harris are among the many people Giaever credits with helping him work his way up the ladder at GE, and it was RPI professor Hillard B. Huntington who sparked Giaever's interest in superconductivity.

 

In 1988, Giaever retired from GE and started up his own company, Applied BioPhysics, with GE colleague Charles Keese. Keese is the president and Giaever, now semi-retired, is the chief technical officer. According to the company's website, its mission is to "apply the results of biophysical research to provide practical tools for cell research and drug discovery."

 

Still in Niskayuna

 

Giaever and his wife still live in their house on Antwerp Road, where they raised a son and three daughters. The couple have eight grandchildren. Out of the spotlight for quite some time by 2008, Giaever and 70 other Nobel Science Laureates created some news by endorsing Barack Obama in his run for the presidency.

 

While he still supports Obama, Giaever ruffled some liberal feathers in a March 2009 letter to the president when he, along with more than 100 international scientists, was critical of Obama's stance on global warming. Giaever thinks the case for global warming is overstated, and in 2011 he failed to renew his membership in the American Physical Society because of it, creating quite a stir in the scientific community.

 

"I like Obama, and I support his health care system even though it's quite complicated, but I guess if you're against global warming you have to be a Republican," said Giaever. "Well, basically I'm a Democrat, but I'm not convinced that global warming is a major issue. I resigned from the APS because they said 'the evidence for global warming was incontrovertible.' That means you can't even discuss it, and I think that's disgusting. That makes it like religion."

 

Religion also happens to be a subject where Giaever disagrees even more vehemently with the president.

 

"I'm not a spiritual person," he said. "I look at the evidence and say, 'is it likely that there's a God up there who I can pray to and who will listen to me?' I don't think so, and that is something I feel like I absolutely know."  

 

 


 

 

Reach Gazette reporter Bill Buell at 395-3190 or bbuell@dailygazette.com

 

 

 

 

 

ECIS Software   

 

Latest Software version:
 

The latest ECIS Software version for Windows is v1.2.135:  

  

            http://www.biophysics.com/software/ECIS_Software_v1_2_135.msi

  

 

If you have a 64 bit Mac running OSX 10.6 or later, and wish to do offline analysis, there is an update to v1.2.135.

The install instructions can be found here:

 

            http://www.biophysics.com/software/ECIS_Mac_v1_2_135_Install_Instructions.pdf

 

 

Software Tip

 

Changing the Font on an Exported Graph

 

If you wish to export a graph for use in a paper or presentation, you can change the displayed font and size.

First click the Export button (or Edit | Export Graph) and a separate plot window will appear. From the new window go to File | Export Setup, select 'Fonts' under 'Properties', and you can change the font type and size and then click 'Apply to Figure'. Then you can save the image to a file by selecting File | Save As and selecting a file format (TIFF recommended).  

 Bar Code

   

  

 

 

  

All arrays now ship with a bar code (code 128).  The bar code contains the model number of the array and a unique serial number.  The bar code can be entered manually or automatically with a bar code scanner (available from ABP).   When entering with the bar code scanner, the array type is automatically set in the ECIS software and the serial number and array type are recorded with the ECIS data. The use of bar code requires V 1.2.135 or later of the ECIS software."

 

 

 

 Early Career Mini-Grants

  

Mini Grant Array Image  

 

 

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.   

  

Applied BioPhysics Inc. Announces the ECIS Trans Filter Adapter

  

The new ECISŪtrans Filter adapter enables the continuous measurement of cell monolayer barrier function (TEER) using membrane inserts in multiple wells.   

 

 

The new trans Filter adapter accommodates standard twenty four well membrane inserts from a broad range of manufacturers. The adapter holds up to eight inserts, and with two adapter devices connected to the ECISŪ data acquisition system, up to sixteen filters can be followed independently. Dedicated software presents real-time, continuous measurement of TEER in ohm-cm2.

 

In the standard ECISŪ culture plates, cells are exposed to media only from their apical sides. With the ECISŪ trans Filter adapter, cells on a membrane can access media on both the basolateral and apical sides and thereby more closely experience their in vivo environment.

 

Other ECISŪ applications include measurements of cell migration, endothelial barrier function, extravasation of normal cell layers by metastatic cells, signal transduction, cell-ECM interactions, cytotoxicity, cytopathic effects of viral infections and cell proliferation.

 

The trans Filter array and the ECISŪ measurement system will be on display at the NAVBO Oct. 20 - 24, 2013, Cape Cod, MA and theAmerican Society for Cell Biology, Dec. 14 - 18, 2013, New Orleans, LA. 

 

For technical questions:

 

Dr. Christian Renken

Applied BioPhysics

185 Jordan Road

Troy, NY 12180

Ph: 1-866-301-ECIS (3247)

Fax 518-880-6860

Renken@biophysics.com

   

ECIS Webinar Schedule 2013

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.

All webinars are held at 11:00am EST. To register for a webinar, please go to:
https://appliedbiophysics.webex.com and scroll to the webinar date of interest.    

  
Cell Invasion / Extravasation Assays - 11:00 AM EST
October 22, 2013

 

Automated Cell Migration - 11:00 AM EST
November 12, 2013

 

Barrier Function Assays - 11:00 AM EST
November 26, 2013

 

Real-time Electroporation and Monitoring - 11:00 AM EST
December 10, 2013


For a more detailed description of each webinar, please visit: http://www.biophysics.com/webinar.php

New Publications

  

Mitochondrial DAMPs Increase Endothelial Permeability through Neutrophil Dependent and Independent Pathways. by Shiqin Sun, Tolga Sursal, Yasaman Adibnia, Cong Zhao, Yi Zheng, ...Haipeng Li, Leo E Otterbein, Carl J Hauser, Kiyoshi Itagaki. PloS one (2013) Volume: 8, Issue: 3 
  
Vascular endothelial hyperpermeability induces the clinical symptoms of Clarkson disease (the systemic capillary leak syndrome). by Zhihui Xie, Chandra C Ghosh, Roshni Patel, Shoko Iwaki, Donna Gaskins, ...Celeste Nelson, Nina Jones, Philip R Greipp, Samir M Parikh, Kirk M Druey. Blood (2012) Volume: 119, Issue: 18
  
Angiopoietin-1 requires IQ domain GTPase-activating protein 1 to activate Rac1 and promote endothelial barrier defense. by Sascha David, Chandra C Ghosh, Aditi Mukherjee, Samir M Parikh. Arteriosclerosis thrombosis and vascular biology (2011) Volume: 31, Issue: 11
  
GPR124, an orphan G protein-coupled receptor, is required for CNS-specific vascularization and establishment of the blood-brain barrier. by Mike Cullen, Mohammed K Elzarrad, Steven Seaman, Enrique Zudaire, Janine Stevens, ...Mi Young Yang, Xiujie Li, Amit Chaudhary, Lihong Xu, Mary Beth Hilton, Daniel Logsdon, Emily Hsiao, Erica V Stein, Frank Cuttitta, Diana C Haines, Kunio Nagashima, Lino Tessarollo, Brad St Croix. Proceedings of the National Academy of Sciences of the United States of America (2011) Volume: 108, Issue: 14
  
Neuronal markers are expressed in human gliomas and NSE knockdown sensitizes glioblastoma cells to radiotherapy and temozolomide. by Tao Yan, Kai Ove Skaftnesmo, Lina Leiss, Linda Sleire, Jian Wang, Xingang Li, Per Øyvind Enger. BMC Cancer (2011) Volume: 11, Issue: 1
  
The effect of temperature on the impedimetric response of bioreceptor hosting hydrogels. by Anthony Guiseppi-Elie, Lauren Koch, Stephen H Finley, Gary E Wnek. Biosensors and Bioelectronics (2011) Volume: 26, Issue: 5
  
Benzyl isothiocyanate inhibits oncogenic actions of leptin in human breast cancer cells by suppressing activation of signal transducer and activator of transcription 3. by Su-Hyeong Kim, Arumugam Nagalingam, Neeraj K Saxena, Shivendra V Singh, Dipali Sharma. Carcinogenesis (2011) Volume: 32, Issue: 3  
  
Tryptase activation of immortalized human urothelial cell mitogen-activated protein kinase. Marentette JO, Hauser PJ, Hurst RE, Klumpp DJ, Rickard A, McHowat J. PLoS One. 2013 Jul 29;8(7):e69948. doi: 0.1371/journal.pone.0069948. Print 2013.
 

Endothelial Cell Permeability during Hantavirus Infection Involves Factor XII-Dependent Increased Activation of the Kallikrein-Kinin System Shannon L. Taylor 1, Victoria Wahl-Jensen 2, Anna Maria Copeland 1, Peter B. Jahrling 2, Connie S. Schmaljohn 1* PLoS Pathogens Received February 1, 2013; Accepted May 16, 2013; Published July 18, 2013
 

Effects of a synthetic PEG-ylated Tie-2 agonist peptide on endotoxemic lung injury and mortality

Sascha David , Chandra C. Ghosh , Philipp Kümpers , Nelli Shushakova , Paul Van Slyke , Eliyahu V. Khankin , S. Ananth Karumanchi , Dan Dumont , Samir M. Parikh American Journal of Physiology - Lung Cellular and Molecular PhysiologyPublished 1 June 2011Vol. 300no. L851-L862DOI: 10.1152/ajplung.00459.2010


Effective treatment of edema and endothelial barrier dysfunction with imatinib. Aman J, van Bezu J, Damanafshan A, Huveneers S, Eringa EC, Vogel SM, Groeneveld AB, Vonk Noordegraaf A, van Hinsbergh VW, van Nieuw Amerongen GP. Circulation. 2012 Dec 4;126(23):2728-38. doi: 10.1161/CIRCULATIONAHA.112.134304. Epub 2012 Oct 25.

 

The antiviral cytokines IFN-α and IFN-β modulate parietal epithelial cells and promote podocyte loss: implications for IFN toxicity, viral glomerulonephritis, and glomerular regeneration. Migliorini A, Angelotti ML, Mulay SR, Kulkarni OO, Demleitner J, Dietrich A, Sagrinati C, Ballerini L, Peired A, Shankland SJ, Liapis H, Romagnani P, Anders HJ. Am J Pathol. 2013 Aug;183(2):431-40. doi: 10.1016/j.ajpath.2013.04.017. Epub 2013 Jun 5.

 

Activation of TRPC6 channels is essential for lung ischaemia-reperfusion induced oedema in mice.
Weissmann N, Sydykov A, Kalwa H, Storch U, Fuchs B, Mederos y Schnitzler M, Brandes RP, Grimminger F, Meissner M, Freichel M, Offermanns S, Veit F, Pak O, Krause KH, Schermuly RT, Brewer AC, Schmidt HH, Seeger W, Shah AM, Gudermann T, Ghofrani HA, Dietrich A. Nat Commun. 2012 Jan 31;3:649. doi: 10.1038/ncomms1660

 

Localized RhoA GTPase activity regulates dynamics of endothelial monolayer integrity.Szulcek R, Beckers CM, Hodzic J, de Wit J, Chen Z, Grob T, Musters RJ, Minshall RD, van Hinsbergh VW, van Nieuw Amerongen GP. Cardiovasc Res. 2013 Aug 1;99(3):471-82. doi: 10.1093/cvr/cvt075. Epub 2013 Mar 27.  

  

 ___________________________________________________________________________________

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 renken@biophysics.com. We will announce your article in our newsletter, post it on our website and send you two FREE 8 well arrays! 
 

 

Upcoming Events

Representatives from Applied BioPhysics will be at the following tradeshows and events:    

 


NAVBO
Oct. 20 - 24, 2013

Cape Cod, MA

 

American Society for Cell Biology
Dec. 14 - 18, 2013

New Orleans, LA

 

Society of Toxicology
March 23 - 27, 2014
Phoenix, AZ

 

American Association for Cancer Research
April 5 - 9, 2014
San Diego, CA

 

Experimental Biology
April 26 - 30, 2014
San Diego, CA

 

ATS 2014 International Conference
May 16 - 21, 2014
San Diego, CA 

 
 
Tip of the Month: Choice of program for ECISŪ time course data acquisition

 

The ECISŪ software offers three different modes for time course data collection:

 

*rapid time collection (RTC)

 

*single frequency / time (SFT)

 

*multiple frequency / time (MFT)

 

The first consideration in choosing a particular mode is whether one is monitoring cell responses that are expected to take place very rapidly. Although commonly cell culture experiments involve changes that take place over several minutes to many hours, rapid events may also be monitored with ECISŪ such as signal transduction or the beating of cardiomyocytes. For these, the rapid data acquisition is the best choice as the RTC mode can be set to measure 25 samples per second. This mode is restricted to monitoring a single well at a time and at one frequency.

 

The single frequency mode (SFT) enables measurements of multiple wells at one AC frequency; each well requires approximately 0.5 seconds for the measurement. Normally before using this mode, the researcher will have carried out additional measurements to determine the appropriate AC frequency where the cellular phenomenon to be monitored is best detected.

 

For most ECIS experiments we strongly recommend using the MFT program, as the data collected in this mode allow one to carry out a more thorough analysis of cellular changes. The MFT mode follows multiple wells over a range of AC frequencies. The frequency set called up by the software depends upon the instrument (Z or Z Theta) and the type of array (8 well or 96 well). For example, the default AC frequencies used with the Z Theta instrument monitoring eight well arrays are 67.5, 125, 250, 500, 1K, 2K, 4K, 8K, 16K, 32K and 64K Hz*. Each well selected is automatically monitored at these 11 frequencies, the required time being approximately 1 second for each frequency for each well. So with 8 wells being followed at 11 different frequencies, the time between data points for any given well is 11x8 sec or about 1.5 minutes.   Distinct aspects of cell behavior can be probed using different AC frequencies, and if you are using the Z Theta instrument, MTF data can be modeled to provide quantitative measurement of defined cell parameters.

 

*Note: It is possible to select different AC frequencies for the MFT mode. Simply click on the Acquire tab and scroll down to "Set Scan frequencies". Here one can manually introduce a list of frequencies to be used in the measurement.

 

 
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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