CORE AND FACILITIES

Imaging

The Imaging Core supports the care and use of instrumentation for imaging of live and preserved organs, cells, tissues, as well as plates and arrays. The Core also provides computer software for the manipulation of images and quantification of image features.

The Imaging Core is supported by NEI Core Center Grant P30 EY014801. Please cite this grant number on all publications that make use of this core.

Research Associate Gabriel Gaidosh manages the Imaging Core

Management                 

Research Associate Gabriel Gaidosh manages the Imaging Core

Resources & Location

The core is comprised of three different facilities located on different floors of the McKnight Vision Research Center building:

• Analytic Imaging
• Time-Lapse Microscopy
• Stereo Microscopy

Please click on the links above to go directly to information about each facility.  Please click on the links below to view the availability calendar for the following equipment:

• Leica TCS SP5 Confocal Microscope
• Carl Zeiss Axiovert Time-Lapse Microscope

Additional Information

Please scroll down to view information about all facilities of the Imaging Core.

Imaging – Analytic Imaging

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

Leadership
The Analytic Imaging Facility is co-directed by:

Pedro I.J. Salas, PhD, Associate Professor of Cell Biology & Anatomy
Valery I. Shestopalov, PhD, Assistant Professor of Ophthalmology

Location
The Analytic Imaging Facility is located on the 7th floor of the McKnight building, room 704a and 704b.

Equipment

• Leica TCS SP5 Confocal Microscopy Instrumentation

Click here for availability calendar

NIH grant S10 RR019382 funded the purchase of this instrumentation.

The Leica TCS SP5 system is capable of multiple excitation wavelengths of 405, 458, 476, 488, 496, 514, 543 and 633 nm. Besides the detection system for 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. 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 surrounded with an acrylic chamber enclosure to control environmental factors for live cell imaging.

• Dell Computer with 3.0 ghz Xeon processor, 3 gig RAM, and software

This computer and the Velocity software described below was purchase with an earmarked gift from Estelle and George Rosenfield. 

• Volocity 3D Rendering Software (Improvision Inc., Lexington, MA

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 you 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 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, colocalization measurements, and area measurements.

• HP LaserJet 3700dtn
  

This printer provides for publication quality color prints of images.

• Molecular Dynamics Phosphorimager

An end of year NIH grant administrative supplement to R01 EY012651-S1 funded the purchase of this instrumentation.

The Typhoon 9410 Trio+ phosphorimager is a three-laser base instrument, which is upgradable to 4-laser version given the need in additional blue line excitation will arise in the future. This configuration will work for radioactivity- and 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 multiple –labeled protein 2D gels for differential high-throughput screens.

Imaging –Time-Lapse Microscopy

The Time-Lapse Microscopy Facility supports the care and use of the Carl Zeiss Axiovert 200m and accessories.  This instrumentation provides the capabilities for time-lapse imaging of live cell dynamics. It also provides multi-fluorescence modalities such as quantitative calcium imaging and FRET, and has software add-ons for cell/object counting and mosaic image acquisition.

Leadership
The Time-Lapse Microscopy Facility is directed by Jeffrey L. Goldberg, MD, PhD

Location
The Time-Lapse Microscopy Facility is located on the 4th floor of the McKnight building, room 413E.

Equipment

• Carl Zeiss compound beam-splitter binocular microscope (Axiovert 200m)
• Zeiss MRm monochrome digital camera
• DG4 light source
• Dual-View Splitter
• CoolSnap DS camera

Click here for availability calendar

This microscope (Zeiss #4730129902) is equipped with a 360 degree rotary stage for alignment and also has epi-fluorescent attachments and a lamp housing made for multi-purpose microscopy including light, phase and immunofluorescence.  The camera has 12-bit, 1300X1030 pixel resolution.  It has the dual-view splitter, 2nd camera (coolsnap DS),  DG4 light source for rapid excitation switching, and is capable of multi-fluorescence modalities such as quantitative calcium imaging and FRET, as well as software add-ons for cell/ object counting and mosaic image acquisition.  It is equipped with a wide variety of quality objectives: 10x, 20x, 40x, 63x, and 100x plan APO.  It also has filter cubes for imaging signals from multiple fluorophors including FITC, CFP, Rhodamine, YFP, Cy5, Indo1, and DAPI.

Imaging – Stereo Microscopy

The Stereo Microscopy Facility supports the care and use of microscopy instrumentation for stereo imaging of cells, tissues, and organs. GFP-transgenic, and other fluorescently labeled cells and tissues, can be viewed, manipulated, and imaged using these instruments.

Leadership
The Stereomicroscopy Facility is directed by Victor L. Perez, MD, Assistant Professor of Ophthalmology.

Location
The Stereomicroscopy Facility is located on the 6th floor of the McKnight building, room 606A

Equipment

• Zeiss Stemi SV11 Fluorescence Stereo Microscope
• AxioCam Mrc Camera

This microscope is equipped with 1.6x high optical quality lenses to provide wide-field stereoscopic image. H-50 arc lamp provides adequate source of light to illuminate fluorescence-labeled samples. Two different excitation/ emission (DAPI, FITC) filter pairs allow multiple fluorophores to be visualized and/or captured by the digital camera.

• Axiostar Plus Upright Fluorescence Microscope
• AxioCam Mrc Camera

This microscope is equipped with 5x, 10x, 20x, and 40x objectives.  Three different excitation/ emission (DAPI, FITC, CY3) filter pairs allow multiple fluorophores to be visualized and/or captured by the digital camera.

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