BD Biosciences Pathways High Throughput Live Cell Confocal Imaging System:
This system has been designed as a high throughput microscopy platform that can be used to for wide field or confocal fluorescence screening of multiwell plates with fixed or live cells. The system is equipped with an enclosed incubation system (temperature and CO2 control) and a single port liquid handling system for experiments involving live cells including excitation ratiometric studies such as intracellular calcium measurements with Fura2. The system is equipped with both laser based and image based autofocus for rapid scanning of multiwell format plates. The system accepts multiple format substrates including histological slides, petri dishes, multi-chambered coverslips and multiwell plates up to 96 and 384 wells. Image acquisition and solution delivery are completely automated. The system can be easily programmed to image multiple fields per well in multiple well format plates acquiring up to 4 fluorescence colors per field. For added stability, the sample remains static while the lenses move around from location to location. Illumination for both, wide field and confocal imaging modes is provided by two 100 watt mercury lamps which can be alternated for excitation ratio measuremens. Confocal imaging is performed using a single spinning disk based on the CARV design. For wide field experiments the confocal disk is easily moved out of the light path. The BD-Pathways software includes image analysis modules that can be used for automated segmentation and measurement of the images. In addition, the high throughput image cytometry software “Acapella” from Perkin Elmer is available via the GNL. Acapella includes a library of ready to use analysis modules but the microscopy core staff can write custom made scripts to complement the existing capabilities. Although, high content imaging capabilities, which involves the ability to perform large scale screening of libraries with robotics for sample preparation and large scale image handling, storage and manipulation, is not available at UTMB, this system could serve as the base for future developments in that direction.