Equipment

Figure 1: The Cellomics ArrayScan VTI
shown in current configuration with
automated robotic plate loading capability

The ArrayScan VTI Imaging Cytometer

The Imaging Cytometry Core is located in Lab 1.45 of the Hillman Cancer Center. The equipment centers on a Cellomics ArrayScan VTi imaging cytometer with up to 40X magnification and a 120W mercury-halide white light source capable of excitation from 300nm-700nm and up to 5-color capability. (Please see the following link for some commonly used fluorophores).* Equipment also includes a 1T server for data storage and a dedicated data analysis terminal.

Figure 1 illustrates the current configuration of the instrument. In addition to the ArrayScan the core maintains a Twister robotic plate loader as well as a Zeiss Apotome for pseudo-confocal capability (7uM sectioning).

The instrument is capable of generating over 150 numerical outputs including mean fluorescent intensity in various cellular compartments, number of regions of punctuate fluorescence per cell, and cell size and shape parameters with the use of whole cell or cytoskeletal dyes. The ArrayScan VTI is an automated fluorescence image analysis system with integrated image analysis and data management that together allow the user to perform High Content Screening analysis (HCS). Conventional inverted fluorescence microscopy is applied with robotics to enable fully automated imaging of cells in multi-well cell culture plates. The VTI brings three common technologies--a fluorescent plate reader, fluorescent microscope, and flow cytometer--into a single instrument. The focus of the VTI is to establish event dynamics across populations and subpopulations like a flow cytometer, while incorporating sophisticated image processing to allow for spatial discrimination of fluorescent dyes within the cell. Sophisticated image analysis enables quantification of cellular fluorescence on a large scale to include highly complex phenomenon like morphological changes involving sub-cellular constituents, whole cell morphology, and multi-cellular structures, for example. Biological content is further increased through spatial analyses of targets labeled with fluorescent dyes. The integrated database allows for remote sharing of data and images and features logical in-line operations like automated dose response curve reporting and relationships within reams of cellular data.

*Some commonly used fluorophores compatible with the ArrayScan VTI:

  • Hoechst 33342
  • DAPI
  • FITC
  • GFP, EGFP
  • YOYO-1
  • Alexa 488
  • Calcein
  • BODIPY-FL
  • Rhodamine
  • Cy3
  • Alexa 546
  • DyLight 549
  • Texas Red
  • Alexa 568/594
  • LsyoTracker Red
  • MitoTracker Red
  • DRAQ5
  • Alexa 647/680
  • TOTO3