xCELLigence - University of Hong Kongcgs.hku.hk/.../xCELLigence_APAC_RoadShow_by_Kok_Long.pdf ·...
Transcript of xCELLigence - University of Hong Kongcgs.hku.hk/.../xCELLigence_APAC_RoadShow_by_Kok_Long.pdf ·...
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xCELLigenceLabel-free, real-time cell monitoring technology
Ang Kok Long, PhDAPAC Regional Product Manager, Cell BiologyRoche Diagnostics
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www.xcelligence.roche.com
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xCELLigence - Real-Time Cell AnalyzerPartnership of Roche and ACEA
in cell incubator
E-Plate 96
Gold Microelectrode Array(covers ~ 80% of the areaon the bottom of a well)
RTCA SP system
on benchtop
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xCELLigence - Real-Time Cell Analyzer Principle – Electric Cell Impedance Sensing
Z = Z0
Baseline
Z = Zcell
Impedance
Z = Zcell 2
Impedance doubled
Z = Zcell 3
Impedance further increased
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xCELLigence - Real-Time Cell Analyzer Detect changes in cell number or cell adhesion/morphology
A parameter termed Cell Index (CI) is derived to represent cell status based on the measured electrical impedance.
30 min
4 hr
42 hr
72 hr
Kirstein_SL et al., 2006
H460 cells
Atienza_JM et al., 2005
NIH 3T3 cells
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xCELLigence - Real-Time Cell Analyzer Detect adhesion strength
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xCELLigence - Real-Time Cell AnalyzerKey features and advantages
Label-free detection– Cells are assayed in more physiological conditions– Avoid artifacts that can be introduced by sample handling/preparation– Avoid compound interference (which can be common in optical-based assays)
Non-invasive, real-time monitoring– Enables long term assay of live cells– Use of internal control for each well (before and after treatment)– Less cells required, important for the use of primary cells/disease-relevant cells from patient biopsy
Automated continuous measurement– Enables data collection for the entire duration of the experiment with no hands-on time– More consistent results (by avoiding operator-related variations)
Obtain distinctive cytological profiles of predictive value
Broad range of applications on a single platform
Enable experiments not possible or very difficult to perform with other technologiese.g. Long time-course, rapid kinetics and long-term effect, multiple sequential treatments
Capture the whole picture and don’t miss the effect you want to analyze!
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xCELLigence - Real-Time Cell AnalyzerCell Index is a quantitative measure of cell number
Cell number 16,000 4,000 1,000 250 125
Cell index 2.723 0.759 0.121 0.025 0.014
SD 0.103 0.04 0.011 0.004 0.003
CV % 3.8 6.0 8.8 15.2 24.8
RTCA Assay MTT Assay
y = 0.0002x + 0.0046R2 = 0.995
0
0.5
1
1.5
2
2.5
3
0 4000 8000 12000 16000
Number of Cells
Cel
l Ind
ex
y = 8E-05x - 0.0099R2 = 0.9958
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0 4000 8000 12000 16000
Number of Cells
MTT
Uni
t
Xing_JZ et al., 2005
NIH-3T3
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xCELLigence - Real-Time Cell AnalyzerCell Index is a quantitative measure of cell number
Hela cells
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xCELLigence - Broad Applications Major application areas
Cell culture management and QC– Cell growth curve, cytological profiles
Comprehensive cytotoxicity assays– Compound & RNAi screening– Cell-mediated cytotoxicity– Virus-mediated cytotoxicity
• virus titration, virus neutralization• minimize exposure risk to pathogen
– Toxicology• Cancer drug discovery in pharma
– in vitro predictive toxicology• Environment health
– Agents with unknown MOA– Profiling tumor cell response to treatment
• Less cells required, important for use of primary cells/patient biopsy
Monitoring cellular signaling/event– Cell proliferation, cell death– Cell differentiation– Cell adhesion, migration and invasion– Receptor activation/inhibition
(e.g. GPCRs, RTKs)
Facilitate cell-based assay development– Shorten assay development time– Simplify optimization of assay conditions– Define when to do what objectively
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xCELLigence – Cytotoxicity AssaysCompound-mediated cytotoxicity
Xing_JZ et al., 2006
Cytostatic
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xCELLigence – Cytotoxicity AssaysCell-mediated cytotoxicity
Zhu et al., 2006
NK : Target ratio
NK-92A549
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xCELLigence – Cytotoxicity AssaysVirus-mediated cytopathic effects (CPE)
MDCK cells + H1N1 virus MDCK cells + H1N1 virus + antiserum
Virus titration Virus neutralization
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Neural Stem Cell (NSC)Shp2 is required for self-renewal and proliferation of NSCs
Ke et al., MCB (2007)
Control: Normal NSCMutant: Shp2-deficient NSC
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Capture the whole picture and don’t miss any effect!Knockdown of DNMT3B4/2 inhibits cell growth and promotes apoptosis
Wang et al., Cancer Research (2007)
H1299 cells
Short term response
Long term response
Dose-dependent response
Target specificity
Flow cytometry⇓
Apoptosis
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Gene OverexpressionFunctional consequence of exogenous wild-type and mutant PHOX2B
Raabe et al., Oncogene (2008)
SK-N-AS cells NLF cells
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Real-Time Assessment of CytotoxicityEasier, less laborious, time-dependent IC50 values
Ceriotti et al., Sensors and Actuators B (2006)
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xCELLigence - Real-Time Cell AnalyzerImprove cell-based assay workflow
Results
Seed cell Treatment
Cell QC(visual inspection with microscope)
Endpoint measurement(sample handling/preparation)
Endpoint assay workflow (step-wise)
‘Black Box’ Data analysis(with third party software)
Automated real-time continuous measurements & data analysis
Seed cell Treatment
Results
xCELLigence workflow (seamless)
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xCELLigence - Real-Time Cell Analyzer Distinctive cytological profiles
Atienza_JM et al., 2006
A new way to look at cell analysis
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xCELLigence SystemKey Product Launches 2008/2009
Q4 08Q3 08 Q1 09
Jul
Nov
RTCA SPSP (1x96) stationW 380 analyzer SP SW, laptop96-well E®- Plate
RTCA MPMP (6x96) stationW 380 analyzer MP SW, laptop96-well E®- Plate
RTCA DPDP (3x16) int. stationSW, laptop16-well E®- Plate
16-well CIM Device
Feb
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xCELLigence – Entry into CytomicsRAS strategy - Complete Solution Provider Inspiring Discovery
CytomicsProteomicsGenomics
Microarray Sequencing
Cells OrganismsOrgans Commumities
2D-GE
Subcellular
CellsDNA, RNA
FC IHC/ISHMS
Proteins
Function
NA Isolation Protein Isolation
RT-PCR
Cell Isolation
Tissues
Sam
ple
Targ
etD
etec
tion
Molecular Profiling Cytological Profiling
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xCELLigence systemAn essential tool in labs working with cells
Just Add Cells !
Simple maintenance-free system, easy to set-up and use
Direct quantitative measurement, no sample handling/preparation required
One For All !
Broad range of applications on one platform, with intuitive integrated software
Greater Insight, True Understanding !
Non invasive, real-time continuous monitoring of live cell assays over long duration
Capture the entire experiment and get the whole picture
Label-free, compatible with many other assays
Obtain Results Not Possible With Endpoint Assays !
Distinctive cytological profile - QC, troubleshooting, cell response profiling predictive of MOA
Improve research quality, productivity and inspiring discovery
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Greater Insight, True Understandingwww.xcelligence.roche.com