High-Throughput Microscopy

 

The Columbia Genome Center is equipped with the IN Cell Analyzer 2000 (GE Healthcare), a high-throughput imaging system for performing high-content analysis.

The instrument is equipped with a large CCD camera (resolution 2048 x 2048 pixels) that is capable of whole-well imaging in 96- or 384-well microplates. The IN Cell 2000 has several imaging modes, including image restoration and deconvolution. This enables users to get confocal-like quality in a high-throughput experimental setting. The instrument is also configured with environmental and temperature control and liquid handling modules to conduct real-time live cell imaging.

High-content image analysis software (IN Cell Investigator) is also integrated into this platform. It includes several image analysis modules (Object Intensity, Nuclear Trafficking, Plasma Membrane Trafficking, Granularity, Cell Cycle Trafficking, Morphology, Dual Object, Neurite Outgrowth, Micro Nuclei, Multi-Target Analysis) which can be implemented either online during image acquisition or after a run is complete. The software uses the graphing and filtering tools of Spotfire Decision Site and allows for up to four independent users to perform the analysis simultaneously.

The IN Cell Analyzer 2000 is integrated with Cytomat microplate managing stations via a Mitsubishi robotic manipulator. This enables users to perform high-throughput screens of up to 180 microplates per set.

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Imaging Synapses Using Fluorescent Small Molecule Probes

Imaging synapses

In a test of the Columbia Genome Center's high-content microscopy system, computational image analysis confirmed a high degree of colocalization of green fluorescent protein-labeled synapsin and the dye FM4-64. The researchers plan a high-throughput screen to identify fluroescent small molecules capable of targeting synapses.

Synapses mediate communication between neurons in the brain, making them critical components for neurological activity. Research has shown that synaptic loss and dysfunction play roles in a number of debilitating brain disorders — including Alzheimer’s disease, major depressive disorder, and autism — but currently no effective method exists for identifying and imaging individual synapses in living human brains. Being able to locate and quantify synapses in patients could greatly improve the diagnosis and monitoring of disease, and potentially offer new approaches for treatment.

Clarissa Waites, an assistant professor of pathology and cell biology at the Columbia University College of Physicians & Surgeons, and Dalibor Sames in the Columbia University Department of Chemistry, have recently embarked on a collaboration with the Columbia Genome Center High-Throughput Screening Facility with the goal of identifying small fluorescent molecules that can selectively localize to synapses. If successful, this project could for the first time provide a method for targeting and imaging synapses in the living human brain.

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