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Advanced
Robotic Health Care Area
Automated Robotically-Based High-Throughput
Radiation Biodosimetry
(funded by NIAID/NIH,
Dr. David Brenner of the Center of Radiological Research, Columbia
University Medical Center is the PI)
Mass radiological triage will be critical after
a large-scale event because of the need to identify, at an early stage,
those individuals who will benefit from medical intervention, and those who
will not. Our goal is to develop a fully automated ultra-high throughput
radiation biodosimetry workstation, using purpose-built robotics and
advanced high-speed automated image acquisition. Maximum throughput will be
30,000 samples / day, compared with throughputs in current devices of a few
hundred samples / day.
The basic system involves the well-characterized
micronucleus assay in lymphocytes, with all the assays being carried out
in-situ in multi-well plates. A key option of the system will be that each
lymphocyte sample will be split in two, with one of the two split samples
being irradiated to a harmful dose level (say, 1.8 Gy), before being
analyzed. This will allow a positive control for each individual, providing
an internal calibration to take into account inter-individual variability in
radiosensitivity. By calling up pre-programmed options in timing, liquid
handling, and image analysis, the device will also measure gamma-H2AX foci
yields, and micronucleus yields in reticulocytes, both providing "same-day
answer" dose estimates. By calling up pre-programmed options in liquid
handling steps, the device will also measure micronuclei in other
readily-accessible tissues, such as exfoliated cells from urine or buccal
smears. |
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Robotic Preparation of cDNA from Single
Cells:
(Funded by NIH, with Dr. Lou Cleveland of
St. Luke's-Roosevelt Hospital Center, New York)
The current state of the art for optical
microscopy of living cells provides an array of techniques of extraordinary
power. Living cells can now be studied in multiple dimensions (three spatial
dimensions, time, multiple wavelengths, and multiple stage positions). A
unique feature of microscopic approaches is the capability of observing
transient, ephemeral structures and interactions on a cell-by-cell basis.
This includes the ability to monitor subcellular processes and to follow
cell movements as well as cell-cell interactions over time. An instrument
that couples the power of multidimensional microscopy with that of DNA array
technology has been developed. Specifically, the instrument in which
individual cells selected on the basis of optically detectable features at
critical time points in dynamic processes can be rapidly and robotically
micromanipulated into reaction chambers to permit amplified cDNA synthesis
and subsequent array analysis. In this way, "snapshots" of gene expression
in single cells can be related to information obtained with multidimensional
microscopy. The proposed instrument incorporated an inverted research
microscope capable of widefield deconvolution microscopy as well as a
robotic system for manipulation of cells and reagents.
Algorithms have been developed for automatic
recognition and manipulation of cells, a requirement for high throughput. In
particular, we developed automatic detection of unstained viable cells in
bright field images using a Support Vector machine with an improved training
procedure; innovative preprocessing approaches to cell recognition, such as
Fisher’s linear discrimination; multiclass cell detection with ECOC
probability estimation. |
