High-throughput cell manipulation and quantitative imaging

Institute For Systems Biology, Seattle WA

Investigators

Abstract

[unreadable] DESCRIPTION (provided by applicant): Few instruments can claim to have the potential to revolutionize basic cell biology experimentation - the LEAP instrument is one. LEAP is an integrated platform which incorporates three core processes essential to all cell-biological/biomedical research: imaging, transfection, and cell purification. These operations can be performed at high speed, with single cell precision, and in a standard microtiter plate format. Due to its optical design which uses a wide angle F-theta lens rather than a microscope objective, the LEAP instrument provides equivalent resolution and significantly higher imaging throughput (up to 105 cells per second), than currently available "high-content screening" systems such as the Zeiss/Cellomics platform. In addition, the LEAP technology uses short intense pulses of visible-wavelength laser light to introduce molecules (DNA, RNA, proteins, dyes, chemicals, and even particles) into a wide variety of mammalian cells at rates up to 103 cells per second. This "optoinjection" process is benign and typically more efficient than current methods to transfect even primary human cells. The instrument includes sophisticated image processing algorithms which allow the automated selection and subsequent targeting of single cells for optoinjection, even in confluent monolayers. The capacity to selectively and efficiently transfect individual cells is a powerful innovation that is unmatched by any existing technology and opens up numerous avenues of research that were previously challenging, if not impossible. Finally, LEAP has the revolutionary capability to purify cell populations. At higher power, the same laser used for optoinjection can be used to selectively ablate cells from a population based on an optical phenotype (fluorescence, morphology, etc.) This permits any user-selected subpopulation of cells to be eliminated, thereby purifying the remaining population for subsequent analysis. This purification capability enables the generation of homogeneous cell populations, which may be sensitively measured by standard biochemical assays, while avoiding the confounding effects of noise introduced by contaminating cells. No other technology currently available offers this level of control over the manipulation of cell lines or primary cells. This machine will significantly accelerate the pace of biomedical research by automating many of the currently laborious and slow techniques of cell biology. The instrument will have a particularly strong impact on those aspects of biomedical research which require intensive study of cells derived from human tissues or blood which are generally difficult to obtain and work with, yet provide the most important insights into the mechanisms of disease. [unreadable] [unreadable] [unreadable]