Real Time Cell Analysis
The Acea Biosciences xCELLigence RTCA DP instrument, (formerly marketed by Roche Applied Science) analyses cell functions in real time using non-destructive, reagent free impedance measurements. Real time monitoring detects short term events which endpoint assays can easily miss. The continuous monitoring of cells can determine the best time perform a step like adding reagents to the cells, or sampling cells for measurements of gene expression.
The reproducibility of the data between experiments tells users if their cells are growing as expected. 5000 to 25000 cells per well is the usual number of cells used.
The cell invasion plate uses Boyden chambers to measure cell migration. For standard plates, an E-plate insert separates one group of cells from another, so that the effects of growth factors secreted by the first group of cell on the second group can be measured. One application is observing a reduction in cell migration long before cell death was seen.
Our instrument can measure a maximum of 48 wells, in 3 plates of 16 wells, which can be run independently. Locally Dr. Eusebio Pires, Dept. Cell Biology, has measured complement mediated antibody toxicity, and Kevin d'Auria, Dept. Biomedical Engineering, has measured cytotoxicity.
Some applications are:
- cell proliferation and doubling times
- cell adhesion and spreading
- cell mediated antibody dependent cell cytotoxicity
- cyotoxicity including IC-50 determination
- receptor signalling such as G-protein coupled receptors
- NK cell mediated cytolysis
- virus mediated cytopathogenicity
- siRNA effects
- monitor cell invasion and migration, using CIM plates
- Cell cell interactions using E-Plate inserts
- monitoring of short term gene expression in conjunction with RT-PCR (realtime cell lysis kit app)
- Inhibition of cell attachment
More applications and information are listed on the manufacturer's
website at:
http://www.aceabio.com/case.aspx
including a list of
320 papers as of 2012 using
the instrument.
There is some background on the instrument here, and some reference documents
for users.
If you are interested in using this instrument, contact John Shannon, jds1c@virginia.edu, ph. 434.243.9399.
Before using human cells or other materials covered by biosafety requirements, you must supply IBC number to John Shannon and verify that your training is current. Users are expected to wear lab coats and gloves.
