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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

Screenshots of key software functionality. (a) The default software page showing the various page tabs; (b) highlighting how to switch between the Cell Index, Normalised Cell Index and the Delta Cell Index view in the Plot tab.
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biosensors-05-00199-f010: Screenshots of key software functionality. (a) The default software page showing the various page tabs; (b) highlighting how to switch between the Cell Index, Normalised Cell Index and the Delta Cell Index view in the Plot tab.

Mentions: The software version used in these experiments is 1.2.1. The software is relatively easy to use and reasonable intuitive for the beginner. The software provides an electronic record of the experimental details. All experimental data in the file are permanent and cannot be altered or changed by the user, which is an excellent aspect to the design. The default open page of the current software has 8 tabs (Exp Notes, Layout, Schedule, Cell Index, Plot, Well Graph, Data Analysis and Message; see Figure A1a). The Exp Notes page is an electronic lab book. We recommend to all new users to complete this section as fully as possible. Layout is the design of the plate (e.g., cells/treatments). The Layout page is important, as the information entered here is used for the grouping of wells for calculating averages (±standard deviation) and subsequent pharmacological plots for calculating EC50 or IC50 values, which is done in the “Data Analysis” tab. The Schedule page is for the timing of the recordings. The recording interval and duration of the experiment will of course vary; but any permutation of timing is possible with the software, and emphasis should be placed on optimally recording the desired cellular response. The Plot tab is useful for visualisation of the experiment in real time. This function is extremely important as the data can be viewed as it happens, which allows xCELLigence to be run in conjunction with other end-point assays and, thus, reveal precisely when the response has occurred. In addition, the Plot tab produces publication quality graphs, and we prefer the simplicity and temporal nature of this output for most of our data. The Well Graph tab provides a snap-shot image of the entire dataset in E96 well format. The Data Analysis tab allows for more advanced pharmacology to be conducted (e.g., EC50/IC50). Finally, the Message tab alerts the user of any errors or faults and records the timing when the plate is engaged or removed from the system. Again, this record is permanent and reveals any warning signals or problems (connection issues, plate missing during sweeping), should they arise.


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)

Screenshots of key software functionality. (a) The default software page showing the various page tabs; (b) highlighting how to switch between the Cell Index, Normalised Cell Index and the Delta Cell Index view in the Plot tab.
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f010: Screenshots of key software functionality. (a) The default software page showing the various page tabs; (b) highlighting how to switch between the Cell Index, Normalised Cell Index and the Delta Cell Index view in the Plot tab.
Mentions: The software version used in these experiments is 1.2.1. The software is relatively easy to use and reasonable intuitive for the beginner. The software provides an electronic record of the experimental details. All experimental data in the file are permanent and cannot be altered or changed by the user, which is an excellent aspect to the design. The default open page of the current software has 8 tabs (Exp Notes, Layout, Schedule, Cell Index, Plot, Well Graph, Data Analysis and Message; see Figure A1a). The Exp Notes page is an electronic lab book. We recommend to all new users to complete this section as fully as possible. Layout is the design of the plate (e.g., cells/treatments). The Layout page is important, as the information entered here is used for the grouping of wells for calculating averages (±standard deviation) and subsequent pharmacological plots for calculating EC50 or IC50 values, which is done in the “Data Analysis” tab. The Schedule page is for the timing of the recordings. The recording interval and duration of the experiment will of course vary; but any permutation of timing is possible with the software, and emphasis should be placed on optimally recording the desired cellular response. The Plot tab is useful for visualisation of the experiment in real time. This function is extremely important as the data can be viewed as it happens, which allows xCELLigence to be run in conjunction with other end-point assays and, thus, reveal precisely when the response has occurred. In addition, the Plot tab produces publication quality graphs, and we prefer the simplicity and temporal nature of this output for most of our data. The Well Graph tab provides a snap-shot image of the entire dataset in E96 well format. The Data Analysis tab allows for more advanced pharmacology to be conducted (e.g., EC50/IC50). Finally, the Message tab alerts the user of any errors or faults and records the timing when the plate is engaged or removed from the system. Again, this record is permanent and reveals any warning signals or problems (connection issues, plate missing during sweeping), should they arise.

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