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Tip of the Month - Don't Overlook Micromotion

An often overlooked aspect of ECIS time-course data is micromotion. This term was coined in a 1991 PNAS paper by Giaever and Keese to describe the fluctuations in impedance observed in confluent cell layers. As was pointed out in that early publication, the source of these variations in the impedance is thought to be due to cellular motions. Of interest is the fact that motions that influence impedance are not only in the XY plane of the electrode but also in the Z direction, as current must flow in the narrow spaces between the basal membrane of the cell and the electrode surface. Since these cellular motions are random in nature, they are most easily seen in a small population of cells and show up with more intensity on 1E arrays than on other arrays where the activities of more cells are averaged.

The graph shows normalized resistance data taken at 4000Hz of two identical cell layers but in one case the cells being monitored are on a single 250 micrometer diameter electrode (1E array) and in the other, on 40 of these small electrodes (10E+ array). Since the data is divided by its starting value both of these traces originate at 1 at time zero. About two hours into the experiment, the cells respond to the addition of fresh medium. Notice that although the overall effect is approximately a 25-30% increase in resistance, the 1E well monitoring only about 50-100 cells shows very pronounced fluctuations in resistance - micromotion.

We have found that micromotion is prevalent in all cells examined with ECIS and seems to be especially pronounced in endothelial cell layers. In addition, in toxicology experiments micromotion can be considerably enhanced at low doses of some toxic compounds.

You may wish to consider measurements of this interesting behavior in your research involving ECIS.

ECIS Application Webinar Series

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

Real-time Electroporation and Monitoring - March 3, 2015

Cell Attachment and Spreading Measurements - March 17, 2015

Signal Transduction Assays - April 7, 2015

Toxicology with ECIS - April 21, 2015

ECIS Theory - May 5, 2015

ECIS Software

Latest Software:

The latest version of the software is v1.2.188 available from:

http://www.biophysics.com/software/ECIS_Software_v1_2_188.msi

Or always get the latest version from:

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

You can check the latest version by going to the Help menu, Check for Updates.

Software Tip:

Please note that in the latest versions the Acquire menu has been reorganized to be more categorized and streamlined.

We recommend that all ECIS instruments and computer systems are connected to a Uninterruptible Power Supply (UPS) to minimize issues with power outages and other electrical noise that may affect your experiment.

If you have issues with the USB connection between the PC and ECIS, often known as the Red Screen, please ensure all power save modes are disabled, including USB suspend. Also try connecting to a different USB port, and don't use a USB hub. Finally, disable webcams, WiFi and internet access during an experiment, and turn off Automatic Windows Updates.

Tradeshows & Events

Therapeutic and Diagnostic of Inflammation,

Cancer and Host Defense of the Gastrointestinal Tract

March 18 - 19, 2015
Petit Science Center
Georgia State University

SOT 2015 Annual Meeting
March 22 - 26, 2015
San Diego, CA

Experimental Biology 2015
March 28 - April 1, 2015
Boston, MA

AACR Annual Meeting 2015
April 18 - 22, 2015
Philadelphia, PA

ATS 2015 International Conference
May 15 - 20, 2015
Denver, CO

ASCB 2015 Annual Meeting
Dec 12 - 16, 2015
San Diego, CA

Recent Publications

Protein tyrosine kinase 6 mediates TNFα-induced endothelial barrier dysfunction. Haines RJ, Beard RS, Wu MH. Biochem Biophys Res Commun. 2015;456(1):190-196. doi:10.1016/j.bbrc.2014.11.057.

Fragmental Frequency Analysis Method to Estimate Electrical Cell Parameters From Bioimpedance Study. Das D, Kamil FA, Agrawal S, Biswas K, Das S. IEEE Trans Instrum Meas.2014;63(8):1991-2000. i:10.1109/TIM.2014.2303553.

Dihydroartemisinin inhibits vascular endothelial growth factor-induced endothelial cell migration by a p38 mitogen-activated protein kinase-independent pathway. Guo L, Dong F, Hou Y, et al. Exp Ther Med. 2014;8(6):1707-1712. doi:10.3892/etm.2014.1997.

A Circuit for Cell-Culture Assays. Huertas G, Maldonado A, Yufera A, Rueda A, Huertas JL IEEE Trans Circuits Syst II Express Briefs. 2014;PP(99):1-1. doi:10.1109/TCSII.2014.2386233.

Limited role of nuclear receptor Nur77 in Escherichia coli-induced peritonitis. Hamers AAJ, Uleman S, van Tiel CM, et al. Infect Immun. 2014;82(1):253-264. doi:10.1128/IAI.00721-13.

Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements. Stolwijk JA, Matrougui K, Renken CW, Trebak M. Pflugers Arch. 2014. doi:10.1007/s00424-014-1674-0.

Loss of cystic fibrosis transmembrane conductance regulator impairs lung endothelial cell barrier function and increases susceptibility to microvascular damage from cigarette smoke. Brown MB, Hunt WR, Noe JE, et al. Pulm Circ. 2014;4(2):260-268. doi:10.1086/675989.

African Trypanosome-Induced Blood-Brain Barrier Dysfunction under Shear Stress May Not Require ERK Activation. Sumpio B, Chitragari G, Moriguchi T, et al. Int J Angiol. 2014. doi:10.1055/s-0034-1370890.

TWEAK/Fn14 pathway is a novel mediator of retinal neovascularization. Ameri H, Liu H, Liu R, et al. Invest Ophthalmol Vis Sci. 2014;55(2):801-813. doi:10.1167/iovs.13-12812.

Mizoribine inhibits the proliferation of renal stem/progenitor cells by G1/S arrest during renal regeneration. Horimoto N, Kitamura S, Tsuji K, Makino H.

Acta Med Okayama. 2014;68(1):7-15. ttp://www.ncbi.nlm.nih.gov/pubmed/24553483. Accessed January 5, 2015.

Effect of the knockdown of death-associated protein 1 expression on cell adhesion, growth and migration in breast cancer cells. Wazir U, Sanders AJ, Wazir A, et al. Oncol Rep. 2014. doi:10.3892/or.2014.3686.

Evidence of a common mechanism of disassembly of adherens junctions through Gα13 targeting of VE-cadherin. Gong H, Gao X, Feng S, et al.

J Exp Med. 2014;211(3):579-591. doi:10.1084/jem.20131190.

Bone morphogenetic protein 2: a potential new player in the pathogenesis of diabetic retinopathy. Hussein KA, Choksi K, Akeel S, et al. Exp Eye Res. 2014;125:79-88. oi:10.1016/j.exer.2014.05.012.

Reduced GM1 ganglioside in CFTR-deficient human airway cells results in decreased β1-integrin signaling and delayed wound repair. Itokazu Y, Pagano RE, Schroeder AS, O'Grady SM, Limper AH, Marks DL. Am J Physiol Cell Physiol. 2014;306(9):C819-C830. doi:10.1152/ajpcell.00168.2013.

Experimental tools to monitor the dynamics of endothelial barrier function: a survey of in vitro approaches. Wegener J, Seebach J.

Cell Tissue Res. 2014;355(3):485-514. doi:10.1007/s00441-014-1810-3.

Folliculin controls lung alveolar enlargement and epithelial cell survival through E-cadherin, LKB1, and AMPK. Goncharova EA, Goncharov DA, James ML, et al. Cell Rep. 2014;7(2):412-423. doi:10.1016/j.celrep.2014.03.025.

PDLIM2 regulates transcription factor activity in epithelial-to-mesenchymal transition via the COP9 signalosome. Bowe RA, Cox OT, Ayllón V, et al. Mol Biol Cell. 2014;25(1):184-195. doi:10.1091/mbc.E13-06-0306.

Differential apicobasal VEGF signaling at vascular blood-neural barriers. Hudson N, Powner MB, Sarker MH, et al.Dev Cell. 2014;30(5):541-552. doi:10.1016/j.devcel.2014.06.027.

Comparison of Markers and Functional Attributes of Human Adipose-Derived Stem Cells and Dedifferentiated Adipocyte Cells from Subcutaneous Fat of an Obese Diabetic Donor. Watson JE, Patel NA, Carter G, et al. . Adv wound care. 2014;3(3):219-228. doi:10.1089/wound.2013.0452.

In vitro and in vivo effects of suppressor of cytokine signalling 7 knockdown in breast cancer: the influence on cellular response to hepatocyte growth factor. Sasi W, Ye L, Jiang WG, Sharma AK, Mokbel K.

Biomed Res Int. 2014;2014:648040. doi:10.1155/2014/648040.

Assessing in vitro cytotoxicity of cell micromotion by Hilbert-Huang Transform. Lai Y-T, Lo C-M. In: 2014 36th Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE; 2014:3200-3203. doi:10.1109/EMBC.2014.6944303.

Interferon consensus sequence-binding protein (ICSBP) promotes epithelial-to-mesenchymal transition (EMT)-like phenomena, cell-motility, and invasion via TGF-β signaling in U2OS cells. Sung JY, Park S-Y, Kim JH, et al. Cell Death Dis. 2014;5:e1224. doi:10.1038/cddis.2014.189.

ICAM-2 regulates vascular permeability and N-cadherin localization through ezrin-radixin-moesin (ERM) proteins and Rac-1 signalling. Amsellem V, Dryden NH, Martinelli R, et al. . Cell Commun Signal. 2014;12(1):12. doi:10.1186/1478-811X-12-12.

Targeting intestinal inflammation with CD98 siRNA/PEI-loaded nanoparticles. Laroui H, Geem D, Xiao B, et al. Mol Ther. 2014;22(1):69-80. doi:10.1038/mt.2013.214.

Central role for hydrogen peroxide in P2Y1 ADP receptor-mediated cellular responses in vascular endothelium. Kalwa H, Sartoretto JL, Martinelli R, et al.

Proc Natl Acad Sci U S A. 2014;111(9):3383-3388. doi:10.1073/pnas.1320854111.

Stimulated bronchial epithelial cells release bioactive lysophosphatidylcholine 16:0, 18:0, and 18:1. Zhuge Y, Yuan Y, van Breemen R, et al. Allergy Asthma Immunol Res. 2014;6(1):66-74. doi:10.4168/aair.2014.6.1.66.

Endotoxin-induced monocytic microparticles have contrasting effects on endothelial inflammatory responses. Wen B, Combes V, Bonhoure A, Weksler BB, Couraud P-O, Grau GER. PLoS One. 2014;9(3):e91597. doi:10.1371/journal.pone.0091597.

Sphingosine-1-phosphate receptor 3 mediates sphingosine-1-phosphate induced release of weibel-palade bodies from endothelial cells. van Hooren KWEM, Spijkers LJA, van Breevoort D, et al. PLoS One. 2014;9(3):e91346. doi:10.1371/journal.pone.0091346.

Caspase-14 expression impairs retinal pigment epithelium barrier function: potential role in diabetic macular edema. Beasley S, El-Sherbiny M, Megyerdi S, et al. Biomed Res Int. 2014;2014:417986. doi:10.1155/2014/417986.

Superparamagnetic iron oxide nanoparticles impair endothelial integrity and inhibit nitric oxide production. Astanina K, Simon Y, Cavelius C, Petry S, Kraegeloh A, Kiemer AK. Acta Biomater. 2014;10(11):4896-4911. doi:10.1016/j.actbio.2014.07.027.

Flavonoids, flavonoid metabolites, and phenolic acids inhibit oxidative stress in the neuronal cell line HT-22 monitored by ECIS and MTT assay: a comparative study. Kling B, Bücherl D, Palatzky P, et al. . J Nat Prod. 2014;77(3):446-454. doi:10.1021/np400518k.

Examining the Cellular Uptake and Toxicity of Engineered Nanomaterials. Kosaraju K, Tarannum M, Crawford S, Garde K, Aravamudhan S.

ECS Trans. 2014;61(36):15-21. doi:10.1149/06136.0015ecst.

Methamphetamine induces autophagy as a pro-survival response against apoptotic endothelial cell death through the Kappa opioid receptor. Ma J, Wan J, Meng J, Banerjee S, Ramakrishnan S, Roy S. Cell Death Dis. 2014;5:e1099. doi:10.1038/cddis.2014.64.

Impact of fibroblast activation protein on osteosarcoma cell lines in vitro. Ding L, Ye L, Xu J, Jiang WG. Oncol Lett. 2014;7(3):699-704. doi:10.3892/ol.2014.1788.

Endothelial barrier protection by local anesthetics: ropivacaine and lidocaine block tumor necrosis factor-α-induced endothelial cell Src activation. Piegeler T, Votta-Velis EG, Bakhshi FR, et al. Anesthesiology. 2014;120(6):1414-1428. doi:10.1097/ALN.0000000000000174.

Evaluation and refinement of a field-portable drinking water toxicity sensor utilizing electric cell-substrate impedance sensing and a fluidic biochip. Widder MW, Brennan LM, Hanft EA, Schrock ME, James RR, van der Schalie WH. J Appl Toxicol. 2014. doi:10.1002/jat.3017.

LRP1 is a receptor for Clostridium perfringens TpeL toxin indicating a two-receptor model of clostridial glycosylating toxins. Schorch B, Song S, van Diemen FR, et al. Proc Natl Acad Sci U S A. 2014;111(17):6431-6436. doi:10.1073/pnas.1323790111.

Heat shock protein 90 inhibitors prevent LPS-induced endothelial barrier dysfunction by disrupting RhoA signaling. Joshi AD, Dimitropoulou C, Thangjam G, et al. Am J Respir Cell Mol Biol. 2014;50(1):170-179. doi:10.1165/rcmb.2012-0496OC.

Electrical cell-substrate impedance sensing with field-effect transistors is able to unravel cellular adhesion and detachment processes on a single cell level. Susloparova A, Koppenhöfer D, Law JKY, Vu XT, Ingebrandt S. Lab Chip. 2014. doi:10.1039/c4lc00593g.

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!