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Application of xCELLigence RTCA Biosensor Technology for Revealing the Profile and Window of Drug Responsiveness in Real Time.

Kho D, MacDonald C, Johnson R, Unsworth CP, O'Carroll SJ, du Mez E, Angel CE, Graham ES - Biosensors (Basel) (2015)

Bottom Line: In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness.The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay.In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, University of Auckland, Auckland 1023, New Zealand. s.ocarroll@auckland.ac.nz.

ABSTRACT
The xCELLigence technology is a real-time cellular biosensor, which measures the net adhesion of cells to high-density gold electrode arrays printed on custom-designed E-plates. The strength of cellular adhesion is influenced by a myriad of factors that include cell type, cell viability, growth, migration, spreading and proliferation. We therefore hypothesised that xCELLigence biosensor technology would provide a valuable platform for the measurement of drug responses in a multitude of different experimental, clinical or pharmacological contexts. In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness. The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay. In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

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Related in: MedlinePlus

Cell Index vs. Normalised Cell Index. (a,b) show exactly the same dataset, where different densities of endothelial cell were grown for 144 h. The top panel shows the raw Cell Index data, whereas the bottom panel is normalised at 24 h (used here as a hypothetical drug treatment time point). Note the very different shapes of the Cell Index curves following normalisation. As the normalisation function converts all values to a proportion of 1.0, important information is lost in the transformation.
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biosensors-05-00199-f012: Cell Index vs. Normalised Cell Index. (a,b) show exactly the same dataset, where different densities of endothelial cell were grown for 144 h. The top panel shows the raw Cell Index data, whereas the bottom panel is normalised at 24 h (used here as a hypothetical drug treatment time point). Note the very different shapes of the Cell Index curves following normalisation. As the normalisation function converts all values to a proportion of 1.0, important information is lost in the transformation.

Mentions: By far the easiest way to observe the data is using the Plot page. This not only displays the data in real time, but allows grouping of treatments as the mean ± standard deviation. The data can be displayed as several options: (I) as the Cell Index; or (II) the Normalised Cell Index; or (III) the Delta Cell Index (see Figure A1b). The Cell Index is the measure of adhesion across the individual well. The greatest Cell Index recording we have observed has been with astrocytes (Cell Index of 10–15, i.e., very adherent cells). In the absence of cells (media only) or with suspension cells, the Cell Index values will be close to zero. It is useful to know the raw Cell Index of the cells, as this provides a measure of how adherent the cells are. The Normalised Cell Index (NCI) is a manipulation of the data, where a specific time is chosen (i.e., when cells were stimulated), which is then set as 1.0 (100% values) by the software. Then, all values are represented as a proportion of this. NCI is useful to estimate the percentage change in adhesion. Caution must be applied when using the NCI function, as information present in the Cell Index data is removed during manipulation to NCI. This is highlighted in Figure A3.


Application of xCELLigence RTCA Biosensor Technology for Revealing the Profile and Window of Drug Responsiveness in Real Time.

Kho D, MacDonald C, Johnson R, Unsworth CP, O'Carroll SJ, du Mez E, Angel CE, Graham ES - Biosensors (Basel) (2015)

Cell Index vs. Normalised Cell Index. (a,b) show exactly the same dataset, where different densities of endothelial cell were grown for 144 h. The top panel shows the raw Cell Index data, whereas the bottom panel is normalised at 24 h (used here as a hypothetical drug treatment time point). Note the very different shapes of the Cell Index curves following normalisation. As the normalisation function converts all values to a proportion of 1.0, important information is lost in the transformation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493546&req=5

biosensors-05-00199-f012: Cell Index vs. Normalised Cell Index. (a,b) show exactly the same dataset, where different densities of endothelial cell were grown for 144 h. The top panel shows the raw Cell Index data, whereas the bottom panel is normalised at 24 h (used here as a hypothetical drug treatment time point). Note the very different shapes of the Cell Index curves following normalisation. As the normalisation function converts all values to a proportion of 1.0, important information is lost in the transformation.
Mentions: By far the easiest way to observe the data is using the Plot page. This not only displays the data in real time, but allows grouping of treatments as the mean ± standard deviation. The data can be displayed as several options: (I) as the Cell Index; or (II) the Normalised Cell Index; or (III) the Delta Cell Index (see Figure A1b). The Cell Index is the measure of adhesion across the individual well. The greatest Cell Index recording we have observed has been with astrocytes (Cell Index of 10–15, i.e., very adherent cells). In the absence of cells (media only) or with suspension cells, the Cell Index values will be close to zero. It is useful to know the raw Cell Index of the cells, as this provides a measure of how adherent the cells are. The Normalised Cell Index (NCI) is a manipulation of the data, where a specific time is chosen (i.e., when cells were stimulated), which is then set as 1.0 (100% values) by the software. Then, all values are represented as a proportion of this. NCI is useful to estimate the percentage change in adhesion. Caution must be applied when using the NCI function, as information present in the Cell Index data is removed during manipulation to NCI. This is highlighted in Figure A3.

Bottom Line: In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness.The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay.In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, University of Auckland, Auckland 1023, New Zealand. s.ocarroll@auckland.ac.nz.

ABSTRACT
The xCELLigence technology is a real-time cellular biosensor, which measures the net adhesion of cells to high-density gold electrode arrays printed on custom-designed E-plates. The strength of cellular adhesion is influenced by a myriad of factors that include cell type, cell viability, growth, migration, spreading and proliferation. We therefore hypothesised that xCELLigence biosensor technology would provide a valuable platform for the measurement of drug responses in a multitude of different experimental, clinical or pharmacological contexts. In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness. The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay. In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

Show MeSH
Related in: MedlinePlus