Advanced Manufacturing Laboratory    



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.


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.