The Analytic Imaging Facility supports the use and maintenance of instrumentation for scanning laser confocal microscopy and phosphorimaging for radioactively-labelled samples. It also provides software, computers, and technical expertise for processing and analysis of photographic and digital images, and quantification of image features.

Location

The Analytic Imaging Facility is located on the 6th floor of the McKnight building, room 621, and on the fourth floor of the McKnight Building, room 413-B in the case of the Typhoon Trio Scanner/Phosphoimager.

Equipment

• Leica Stellaris 5 Inverted Confocal Microscopy System

The Leica TCS SP5 Stellaris system is capable of super-resolution imaging using multiple excitation wavelengths in the visible light spectrum: 405, 458, 476, 488, 496, 514, 543, and 633 nm. Besides the ultra-sensitive, high-speed detection system allowing for all 4 fluorescence channels and one transmission channel simultaneously, this system is also equipped with an acusto-optical-beam-splitter which provides the highest flexibility for setting up the required experimental parameters. The scope is equipped with 5x, 10x, 20x, 40x, and 63x objectives. This System allows great user flexibility with resolution (up to 64 megapixels) and frame rate (up to 200fps).

Both inverted and upright microscopes are available. The inverted scope is encased into an acrylic environment control chamber designed to control temperature and other environmental factors for prolonged live-cell imaging.

• Dell Precision T7500 Xeon 2.67 Ghz, with 12 gig RAM, and Volocity analysis software. This computer and the Volocity software described below were purchased with an earmarked gift from Estelle and George Rosenfield.
• Volocity 3D Rendering Software (Quorum Technologies, Puslinch, Ontario, Canada)

This software package consists of two parts – Volocity Visualization (rendering part) and Volocity Classification (3D/4D image analysis part). This product provides excellent visualization and editing tools along with great measurement and “feature search” capabilities. It allows researchers to discover, visualize and measure features hidden inside cells or tissues via analysis of images after restoration and rendering.

The Volocity 3D Rendering software is the first true color 4D rendering software (3D plus time) designed for biomedical imaging. It uses new, highly advanced algorithms to provide high-speed easy to use, interactive rendering of time-resolved color 3D image volumes. This software allows volumetric object measurements, quantitative analysis of 4D images, and compiling such images into movies.

• Leica Application Suite
  Leica application suite is useful for processing confocal images. It is capable of exporting image/movie files, photo manipulation (cropping, contrast adjustments, background subtraction), 3d projection/3d deconvolution, intensity measurements, co-localization measurements, and area measurements.
• Molecular Dynamics Typhoon 9410 Trio confocal Scanner/Phosphorimager
  NIH grant administrative supplement to R01 EY012651-S1 funded the purchase of this instrumentation.

The Typhoon 9410 Trio+ scanner/ phosphorimager is a three-laser base instrument. The current configuration researchers to obtain images of radioactivity labeled samples, two-color fluorescence-labeled tissues, 1D and 2D gels, plates, and arrays. The Typhoon is equipped with ImageQuant Image Analysis Software and is optimized with Ettan DIGE and DeCyder Differential Analysis Software for full automatization of proteomics analysis using 2D gels. It can detect multiple types of labeling: fluorescence, chemiluminescence, and radioactivity.

This instrument has more than 30 different applications including Western, Northern, and Southern blots, primer extension and RNAse protection experiments, two-color and one-color microarrays, and 1D and 2D multiple-labeled gels. The Typhoon 9410 is preset for the analysis of multiplex–labeled protein 2D gels for differential high-throughput screens.