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Developing high throughput mammalian expression platforms (1.4 MB)Ben Hughes, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, BNP Conference, 2009 |
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Objective, time-saving analysis
 Save time by analyzing selected wells (highlighted in green). Red wells are excluded.
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Review results and export data
- Dual gating for colony area and compactness
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View each well at different time points
- Export colony data
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View number of colonies per well
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Colonies are identified in well images
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Verification of monoclonality
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Focus only on single colonies – saving time and effort
After initial seeding, CloneSelect Imager can image every well, at any time point, using a ‘loci of growth’ functionality to highlight those wells that contain a single colony – the essential requirement for output from FACS and limiting dilutions. The user only needs to review wells with a single colony.
Confidently verify colony origin
The growth (image) history of each well can be tracked back to its starting point – providing evidence of monoclonality
Data courtesy of CMC ICOS Biologics Inc., USA
Monitor colony growth after selection of secreting cells
After screening and selection of cell lines using ClonePix FL, CloneSelect Imager provides the ideal solution for monitoring and evaluating subsequent outgrowth and, eventually, productivity of the selected cell lines.
Track colony outgrowth
Outgrowth of a single well after 7 days - confluence overlay viewed for each imaging timepoint
Normalize downstream results to cell number
CloneSelect Imager estimates the number of cells per well. Data is then exported to enable normalization of results from ELISA or cell-based assays against cell number.
Correlate protein productivity and growth of cell line
Assessment of cell viability (MTT assay)
CloneSelect Imager offers several benefits over traditional MTT assays for assessing cell viability in response to a cytotoxic challenge.
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Replace time-consuming conventional techniques
- Monitor cell death over time
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Ensure a permanent record of data
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Avoid costly reagents - no staining required
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Images of HeLa cells - wells exposed to different level of cytotoxin
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Dose response curve: decreasing cell confluence with increasing exposure to cytotoxin |
High resolution images show growth of cells exposed to low, medium and high cytotoxin levels
Read more in Publications below
Case Study: Beckman Coulter - Automation of Cell Line Optimization
Presented at LabAutomation 2009 by Dr. Christoph Krüll, Product Manager Automated Solutions, Beckman Coulter Europe.
Read how CloneSelect Imager is utilized in a novel hardware solution for automating cell line optimization and expansion.
Further information
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Beckman Coulter - Cell Culture Automation in BioPharma: Cell Line Optimization and Monoclonal Antibody Production
Case Study: Tecan - Monoclonal antibody production, cell expansion
Read how the Tecan Integration Group integrated CloneSelect Imager with Freedom EVO® to automate cell expansion and confluence measurement of MAb-producing cell lines at Monash University, Australia.
Further information
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Tecan Integration Group - Monoclonal Antibody Production, Cell Expansion. Part of the Monash system.
Case study: Using CloneSelect Imager for Media and Cell Culture Optimization
The NCRIS Biologics Facility frequently works with new cell lines and stable transfectants requiring the development of chemically-defined media to facilitate serum-free recovery wherever possible. In this case study, CloneSelect Imager is used to identify optimal culture conditions for low cell density / clonal outgrowth.
Case Study of CloneSelect Imager - Media and Cell Culture Optimization
Data courtesy of Ben Hughes
Senior Bioprocess Engineer, NCRIS Biologics Facility
Australian Institute for Bioengineering and Nanotechnology (AIBN)
University of Queensland
Acknowledgements to Warren Pilbrough and Dr. Trent Munro
Senior Bioprocess Engineer, NCRIS Biologics Facility
Australian Institute for Bioengineering and Nanotechnology (AIBN)
University of Queensland
Acknowledgements to Warren Pilbrough and Dr. Trent Munro
Objectives
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Monitor serially-diluted cultures in a 96-well format to rapidly screen culture variables
- Tissue culture ware (treated vs. non-treated, square vs. round bottom wells)
- Media (volume, commercial vs. in-house formulations, supplementation)
- Environment (sealed plates, humidification, effect of disturbances)
Workflow
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Identifying low-cell density growth conditions
- Significant improvements achieved in 2-week period with minimal effort by single FTE
Robust and extended growth range achieved over base-case (day-7 data)
Additional information gained on cellular morphology and understanding of growth characteristics
- Multiple nucleation points vs. “edge only” growth
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Wells showing multiple nucleation growth points
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Wells showing edge growth characteristics
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Identified sub-optimal environmental conditions or “edge-effects”
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Plate demonstrating edge effects
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Plate demonstrating environmental effects
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Summary
AIBN conclusions:
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Excellent workflow obtainable using CloneSelect Imager
- Maximize success rate for serum-free colony outgrowth in chemically-defined media by prior optimization of growth conditions
- Efficient tracking of cell confluence ensures high viability is maintained, leading to rapid expansion for evaluating clones in shake flasks (2-3 weeks from picking)
- Use of the CloneSelect Imager is particularly useful for optimizing clonal outgrowth strategies when platform approaches are not suitable i.e., investigating new cell lines or variants
Posters and presentations
Scientific posters and presentations from users of CloneSelect Imager
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Rapid development of a serum-free, suspension-adapted CHO cell production process (462.5 kB)
Ray Davis, Amy Westwood, Dan Rowe, Howard Clarke. CMC ICOS Biologics Inc. |
Publications
A small selection of noteworthy papers and articles.
Accurate non-invasive image-based cytotoxicity assays for cultured cells
BMC Biotechnology 2010, 10:43doi:10.1186/1472-6750-10-43
Abstract
Background
The CloneSelectTM Imager system is an image-based visualisation system for cell growth assessment. Traditionally cell proliferation is measured with the colorimetric MTT assay.
Results
Here we show that both the CloneSelect Imager and the MTT approach result in comparable EC50 values when assaying the cytotoxicity of cisplatin and oxaliplatin on various cell lines. However, the image-based technique was found non-invasive, considerably quicker and more accurate than the MTT assay.
Conclusions
This new image-based technique has the potential to replace the cumbersome MTT assay when fast, unbiased and high-throughput cytotoxicity assays are requested.
Background
The CloneSelectTM Imager system is an image-based visualisation system for cell growth assessment. Traditionally cell proliferation is measured with the colorimetric MTT assay.
Results
Here we show that both the CloneSelect Imager and the MTT approach result in comparable EC50 values when assaying the cytotoxicity of cisplatin and oxaliplatin on various cell lines. However, the image-based technique was found non-invasive, considerably quicker and more accurate than the MTT assay.
Conclusions
This new image-based technique has the potential to replace the cumbersome MTT assay when fast, unbiased and high-throughput cytotoxicity assays are requested.
Brochure
CloneSelect™ Imager
Objective, quantitative assessment of cell growth
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