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Reminder: SWEHSC Investigators (including their staff and students) should contact the Cellular Imaging Shared Service directly for advice regarding experimental design, training, and technical support for this instrument.
Light Microscopic & Image Capture Capabilities:
LSN workstation: The Olympus IMT-2 inverted microscope is equipped with a digital camera that can capture 1280x1024, 10 bit (1024 intensity or grayscale values) images of brightfield, phase contrast, DIC and fluorescence (FITC cube) imaging modes. The microscope has motorized controllers for shutters, microscope filters and x-y-z stage control. The automated stage has holders for microscope slides, 35mm culture dishes and 96 well plates (note: DIC imaging uses polarization and cannot be performed through plastic). A BiopTechs Delta T3 system for keeping 35mm culture dishes at 37°C enables the system to be able to capture time-lapse images of cultured cells over periods of several hours. A copy of Microsoft Excel™ is available on the computer to assist in transferring the numerical data..
The CBA workstation computer was upgraded (larger hard disks, Core Duo processor) in May 2007 and this included an upgrade to the latest version of the SimplePCI software. In addition, a quantitative FRET module was added to the software. This upgrade was paid for out of the Cellular Imaging Shared Service budget.
AZCC workstation: The Arizona Cancer Center workstation is located in room 3964 (see map). This workstation has a SPOT-Jr color digital camera (710x512 pixels) mounted on an upright Nikon T-200 brightfield microscope. The dual-processor workstation has 2GB of RAM and can save data to a 250MB ZIP drive, CD/DVD burner or USB devices.
Image Analysis & Measurement Capabilities:
The workstations use the SimplePCI software (C•Imaging Systems, by Compix, Inc.). Digital images captured from any appropriately calibrated source (e.g. confocal, TEM, SEM, and other microscopes or scanned images) can be analyzed by the software. The software has the capability to measure a large number of parameters related to size, shape, intensity and position. In many instances the software can configured to be acquire measurements in a semi-automated mode. Basic statistical measurements are included in the software. Numerical data can be exported as Microsoft Excel™ spreadsheets.
The software has been used to measure/capture:
• The distance traveled by cultured cells over time (time lapse)
• The length of dendrites (confocal images)
• Calculate pH based on using color ratios from SNARF dyes (Zeiss confocal images)
• Point counting in lung tissue (light microscope images)
• Image montages of whole cross-sections that were later used for 3D reconstruction or poster presentations (light microscope images)
• Create time-lapse movies for presentations (light microscope images)
• Count fluorescently stained nuclei to measure migration in a matrix (fluorescence microscope images)
• Measure sizes of intestinal villi in histologic samples (light microscope images)
• Measure the area of lipid droplets found in Oil Red O stained histologic samples (light microscope images)
• 3D rotations of multi-level/Z-stack images (light microscope images)
• Automatically count the number of cells that crossed the membrane in transwell plates (24 well plates).
• Analysis of changes in melanized skin moles to assess the success of cancer treatment (macro photography).
