omation mbers - Society for Laboratory Automation and Screening

8:30 am Wednesday, February 4 Emerging Technologies – Cell Based Screening Room A3
Jeffrey Price
University of California, San Diego
9500 Gilman Drive
La Jolla, California 92093-0412
price@bioeng.ucsd.edu
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Co-Author(s)
Michael A. Mancini
Baylor College of Medicine
Susanne Heynen, Ivana Mikic, Tim Moran
Q3DM, Inc.
Dynamic Data Mining and High Throughtput Microscopy Improve Productivity of Assay
Design and Screening
Automated high throughput/high-resolution microscopy (0.5 to 0.95 NA lenses) creates a potentially overwhelming
number of cellular and subcellular measurements. Cellular heterogeneity adds complexity, can hinder screen
significance and complicate assay design. Dynamic data-driven mining creates defined subpopulations without
physical sorting to facilitate rapid design and testing of new subcellular assays and decreasing preparation
time. A large suite of cell-by-cell subcellular parameters enables rapid culling together the best fluorescence,
morphometry, translocation and pattern measurements for a new assay. Here, the impact of dynamic data mining
was established in development of a ligand-dependent androgen receptor (AR) trafficking assay. Transiently
transfected GFP-AR and AR619 (an inactivating prostate cancer-associated point mutation) were introduced
into HeLa cells and analyzed with the EIDAQ 100 high throughput microscopy (HTM) system. AR subcellular
distribution was examined for nuclear translocation and subnuclear distribution patterns in response to an
11-point dose response of agonist (R1881) or antagonists (estradiol, Casodex, OH-Flutamide), and demonstrates
the advantage of data mining. AR subpopulations responded consistently to ligand concentration in nuclear
translocation (agonist and antagonist for both AR and AR619). However, only R1881 induced appearance of large
subnuclear foci in AR619. Rapid experimentation with various measurements also enabled assay development
that was sensitive to subtle, ligand-dependent differences in subnuclear pattern of GFP-AR. Cell cycle phases and
metabolic states are additional examples where subpopulation assays may enable more productive screening.
Thus, while image-based subcellular imaging can at first appear to increase assay complexity, powerful dynamic
data mining tools also enable rapid development of new subcellular assays.
8:45 am Wednesday, February 4 Emerging Technologies – Cell Based Screening Room A3
Steve Richmond
Genetix Ltd
Queensway, New Milton BH25 5NN United Kingdom
steve.richmond@genetix.com
Development of a Mammalian Cell Colony Imaging and Picking Robot: ClonePix
Co-Author(s)
Chris Mann, Sky Jiang,
James Colehan, Sarah Stephen,
Julian Burke
Robotics for picking microbial colonies have been available for approximately 10 years and played a key role in
the various genome sequencing projects. Up until now no equivalent systems have existed for handling colonies
of mammalian cells in culture. The task is more challenging as they are more fragile, more prone to contamination
and often grow adhered to a surface. We believe that ClonePix is the first robot to automate transfer of cultured
mammalian cells. The system picks both adherent colonies and those grown in semi-solid media and transfers
them to micro plates. Culture dishes to be processed are imaged using a high-resolution CCD camera. The image
is analyzed and a user-defined clone pick list is generated. The selected clones are picked using a novel 8-channel
head. Each of the channels fires independently, allowing for high throughput picking at rates of ~200 clones per
hour. The location of each clone in the microplate is automatically logged for sample tracking. Sterility of the tips
is achieved via an ethanol wash combined with a high-temperature dryer. External contamination is minimised by
the HEPA filter with positive air pressure and a fully enclosed working area; which can be disinfected with a UV
lamp prior to use. Applications of ClonePix include selection of hybridomas expressing monoclonal antibodies and
selection of transformed cells.
PODIUM ABSTRACTS