A morphological biosensor for mammalian cells

Giæver, Ivar; Keese, Charles R.

doi:10.1038/366591a0

Cited by 707 publications

(475 citation statements)

References 6 publications

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“…Changes in the shape of PBCECs during apoptosis were monitored by electric cell-substrate impedance sensing (Giaever and Keese, 1993;Keese and Giaever, 1994). The method is based on measuring non-invasively the frequency-dependent electrical impedance of cell-covered gold-film electrodes along the time course of the experiment.…”

Section: Electric Cell-substrate Impedance Sensing (Ecis)mentioning

confidence: 99%

“…In the light of this background it was our objective to develop a new bio-analytical assay to monitor apoptosis in cultured cells non-invasively and in real time. As the three-dimensional shape of adherent cells is a very sensitive indicator for their viability or any deteriorating changes in their environment we based the new assay on a shape-sensitive electrochemical technique that is referred to as electric cell-substrate impedance sensing (ECIS) (Giaever and Keese, 1993;Keese and Giaever, 1994). In ECIS the cells are grown on gold-film electrodes that are deposited on the surface of ordinary cell culture dishes.…”

Section: Introductionmentioning

confidence: 99%

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Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Arndt

Seebach

Psathaki

et al. 2004

Biosensors and Bioelectronics

290

191

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Apoptosis is a strictly regulated and genetically encoded cell 'suicide' that may be triggered by cytokines, depletion of growth factors or certain chemicals. It is morphologically characterized by severe alterations in cell shape like cell shrinkage and disintegration of cell-cell contacts. We applied a non-invasive electrochemical technique referred to as electric cell-substrate impedance sensing (ECIS) in order to monitor the apoptosis-induced changes in cell shape in an integral and quantitative fashion with a time resolution in the order of minutes. In ECIS the cells are grown directly on the surface of small gold-film electrodes (d = 2 mm). From readings of the electrical impedance of the cell-covered electrode, performed with non-invasive, low amplitude sensing voltages, it is possible to deduce alterations in cell-cell and cell-substrate contacts. To improve the sensitivity of this impedance assay we used endothelial cells derived from cerebral micro-vessels as cellular model systems since these are well known to express electrically tight intercellular junctions. Apoptosis was induced by cycloheximide (CHX) and verified by biochemical and cytological assays. The time course of cell shape changes was followed with unprecedented time resolution by impedance readings at 1 kHz and correlated with biochemical parameters. From impedance readings along a broad frequency range of 1-10 6 Hz we could assign the observed impedance changes to alterations on the subcellular level. We observed that disassembly of barrier-forming tight junctions precedes changes in cell-substrate contacts and correlates strongly with the time course of protease activation.

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Section: Electric Cell-substrate Impedance Sensing (Ecis)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Arndt

Seebach

Psathaki

et al. 2004

Biosensors and Bioelectronics

290

191

View full text Add to dashboard Cite

show abstract

“…Real‐time cell monitoring using electrical properties (electric cell‐substrate impedance sensing, or ECIS) was introduced by Giaever and Keese over 20 years ago 85. As a label‐free, noninvasive biophysical assay detecting dynamic cell responses, it provides a valuable tool for the investigation of NM toxicity and early stage efficacy testing of NP‐linked drugs.…”

Section: Impedance‐based Monitoringmentioning

confidence: 99%

High throughput toxicity screening and intracellular detection of nanomaterials

Collins

Annangi

Rubio

et al. 2016

WIREs Nanomed Nanobiotechnol

108

104

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With the growing numbers of nanomaterials (NMs), there is a great demand for rapid and reliable ways of testing NM safety—preferably using in vitro approaches, to avoid the ethical dilemmas associated with animal research. Data are needed for developing intelligent testing strategies for risk assessment of NMs, based on grouping and read‐across approaches. The adoption of high throughput screening (HTS) and high content analysis (HCA) for NM toxicity testing allows the testing of numerous materials at different concentrations and on different types of cells, reduces the effect of inter‐experimental variation, and makes substantial savings in time and cost. HTS/HCA approaches facilitate the classification of key biological indicators of NM‐cell interactions. Validation of in vitro HTS tests is required, taking account of relevance to in vivo results. HTS/HCA approaches are needed to assess dose‐ and time‐dependent toxicity, allowing prediction of in vivo adverse effects. Several HTS/HCA methods are being validated and applied for NM testing in the FP7 project NANoREG, including Label‐free cellular screening of NM uptake, HCA, High throughput flow cytometry, Impedance‐based monitoring, Multiplex analysis of secreted products, and genotoxicity methods—namely High throughput comet assay, High throughput in vitro micronucleus assay, and γH2AX assay. There are several technical challenges with HTS/HCA for NM testing, as toxicity screening needs to be coupled with characterization of NMs in exposure medium prior to the test; possible interference of NMs with HTS/HCA techniques is another concern. Advantages and challenges of HTS/HCA approaches in NM safety are discussed. WIREs Nanomed Nanobiotechnol 2017, 9:e1413. doi: 10.1002/wnan.1413For further resources related to this article, please visit the WIREs website.

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“…BLMVEC were grown to confluence in polycarbonate wells containing evaporated gold microelectrodes (surface area 10 Ϫ3 cm 2 ) in series with a gold counter electrode (surface area 1.0 cm 2 ) connected to a phase-sensitive, locked-in amplifier (16). Measurements of transmonolayer electrical resistance (TER) were performed by using an electrical cell-substrate impedance-sensing system (Applied BioPhysics, Troy, NY).…”

Section: Preparation Of Hbmentioning

confidence: 99%

Quantitative assessment of hemoglobin-induced endothelial barrier dysfunction

Dull

DeWitt

Dinavahi

et al. 2004

Journal of Applied Physiology

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ticelli. Quantitative assessment of hemoglobin-induced endothelial barrier dysfunction. J Appl Physiol 97: 1930 -1937, 2004. First published July 23, 2004 doi:10.1152/japplphysiol.00102.2004.-Hemoglobin (Hb)-based O 2 carriers (HBOC) are undergoing extensive development as potential "blood substitutes." A major problem associated with these molecules is an increase in microvascular permeability and peripheral vascular resistance. In this paper, we utilized bovine lung microvascular endothelial cell monolayers and simultaneously measured Hb-induced changes in transendothelial electrical resistance, diffusive albumin permeability, and diffusive Hb permeability (P DH) for three forms of Hb: natural tetrameric human Hb-A and two polymerized recombinant HBOCs containing ␣-human and ␤-bovine chains designated Hb-Polytaur (molecular mass: 500 kDa) and Hb-(Polytaur) n (molecular mass: ϳ1,000,000 Da), respectively. Hb-Polytaur and Hb-(Polytaur)n are being evaluated for clinical use as HBOCs. All three Hb molecules induce a rapid decline of transendothelial electrical resistance to 30% of baseline. Diffusive albumin permeabiltiy increases, on average, approximately ninefold (2.78 ϫ 10 Ϫ7 vs. 2.47 ϫ 10 Ϫ6 cm/s) in response to Hb exposure. All three Hb molecules induce an increase in their own permeability, a process that we have called Hb-induced Hb permeability. The magnitude of change of P DH is also related to Hb size. When PDH is corrected for the diffusive coefficient for each Hb species, no evidence of restricted diffusion is found. Immunofluorescent images demonstrate Hb-induced actin stress fiber formation and large intercellular gaps. These data provide the first quantitative assessment of the effect of polymerized HBOC on their own diffusion rates over time. We discuss the importance of these findings in terms of Hb extravasation rates, molecular sieving, and clinical consequences of HBOC use.albumin; blood substitutes; restricted diffusion HEMOGLOBIN (HB)-BASED O 2 carriers (HBOC) have received considerable attention as blood substitutes, and interest in developing these therapeutic macromolecules remains high, given the shortage and limited shelf-life of banked red blood cells, infection concerns, and, at times, unacceptable delays in crossmatching banked blood (1,24,26,33). Unfortunately, a number of unexpected problems have been associated with the use of HBOC, including extravasation, vasospasm, renal toxicity, and patient death (1,24,33). The ultimate utility of HBOC is to provide adequate oxygen delivery to patients requiring immediate resuscitation, as may occur following major trauma or surgical procedures associated with large-volume blood losses. Two untoward consequences of HBOC are extravasation of the HBOC itself, which is no longer able to participate in oxygen delivery, and the associated vasoconstriction due, at least in part, to local nitric oxide (NO) scavenging. The decrease in plasma Hb and localized vasospasms in the setting of anemia may create regional hypoxia. Overcoming these issues will...

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A morphological biosensor for mammalian cells

Cited by 707 publications

References 6 publications

Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

Bioelectrical impedance assay to monitor changes in cell shape during apoptosis

High throughput toxicity screening and intracellular detection of nanomaterials

Quantitative assessment of hemoglobin-induced endothelial barrier dysfunction

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