Bioimaging Facility

Director - James Keen, Ph.D.

Contact – Manager

215-503-4770

Mission, Goals, Capabilities / The goal of the Kimmel Cancer Center Bioimaging Shared Resource is to provide powerful, reliable, and readily accessible light microscopic image acquisition and analysis capabilities to KCC investigators and Jefferson faculty members. The Bioimaging Shared Resource provides laser point-scanning and spinning disk confocal, TIRF (total internal refection fluorescence), and widefield epifluorescence microscopy capabilities allowing for multi-wavelength visualization and analysis of fixed and living specimens, Z series, single molecule events at surfaces and interfaces, on millisecond to hour timeframes, as well as image analysis and processing expertise.
Major Equipment / 1. Zeiss LSM 510 Meta Confocal Laser Scanning Microscope:
◦Zeiss Axiovert 200M inverted microscope
◦10x, 20x, 40x, 40x oil, 63x oil and 100x oil objectives
◦4 lasers with excitation lines at 405, 458, 477, 488, 514, 543, and 633 nm
◦META detector for resolution of very closely emitting fluorophores
◦DIC transmitted light
◦AIM acquisition and analysis software
◦Programmable X,Y,Z stage for multiple positions and stitching of large areas
◦z-stack, FRET, FRAP, photoactivation, time lapse functions, tiling
◦Temperature-controlled chamber for live imaging
◦Objective warmer
◦Anti-vibration isolation table

NikonC1 Plus laser confocal microscope:

◦Nikon TiE inverted microscope
◦20x, 40x oil, and 60x oil objectives
◦4 lasers with excitation lines at 405, 458, 477, 488, 561, and 640 nm
◦DIC transmitted light
◦EZ-C1 image acquisition and analysis software and Elements image analysis software
◦Z-stack, FRET, FRAP and time lapse functions
◦Anti-vibration isolation table
3. TIRF and Spinning Disk Microscopy
The TIRF (total internal refection fluorescence) microscopy system will permit high sensitivity, high speed imaging of biological phenomena occurring at the ventral surface (≤0.2 µm) of adherent cells without excitation further into the cell interior. Two discrete signals can be monitored simultaneously and a third can be imaged sequentially. The spinning disk confocal head is capable of continuous high-speed high-resolution imaging of live cells. Finally, photobleaching and photoactivation at all wavelengths can be employed in combination with all imaging modalities.
Andor/Nikon TIRF/Spinning Disk Confocal/Widefield Microscope:
◦Nikon TiE inverted microscope with PFS for image stability control
◦100x/1.49 NA TIRF objective
◦Andor laser control with 405, 488, 561 and 640 nm laser lines (all 100 mW and fast switching)
◦DIC transmitted light
◦Nikon TIRF E Motorized Illuminator
◦Yokogawa CSU-X1 spinning disk head for confocal imaging
◦Sutter Lambda DG4-Plus illuminator
◦Two Andor Ion X3 EM-CCD cameras each equipped with Dual-view2 splitters for simultaneous red/green imaging
◦Andor FRAPPA unit for photobleaching and photoactivation (operable under either Andor IQ or MetaMorph software)
◦Ludl programmable X,Y,Z stage with fast piezo Z drive for multiple positions and stitching of large areas
◦Z-stack, FRET, FRAP, photoactivation, time lapse functions, tiling
◦MetaMorph Premier image acquisition and analysis software
◦Okolab stage-top incubator for 35 mm dishes, 6-well plates, and 1" x 3" chamber slides
◦Anti-vibration air table
4. WideField Fluorescence Microscopy
The widefield epifluorescence imaging system is specifically designed for high quality acquisition of live cell images over extended time periods (minutes to hours) with minimal photodamage under a controlled environment. Multiple regions of interest in a single sample can be repeatedly examined. The high speed Lambda DG4 filter changer allows for rapid excitation light changes to provide multiple signal acquisition with high temporal resolution. In addition, brightfield and DIC transmitted light images can be acquired.
◦Zeiss Axiovert 200M inverted microscope
◦10x, 20x, 40x oil, 63x oil and 100x oil objectives
◦Sutter Lambda DG4 illuminator
◦DIC transmitted light
◦Andor Zyla sCMOS camera
◦MetaMorph Premier image acquisition and analysis software
◦Prior Proscan III programmable X,Y,Z stage for multiple positions and stitching of large areas
◦Time lapse functions, tiling
◦Okolab environmental chamber
◦Programmable pump and perfusion chamber
◦Anti-vibration isolation table
Services: / Point-scanning laser confocal microscopy ideal for viewing fixed specimens with thicknesses up to several hundred microns, or brief observation of live samples. Most investigators use these systems, particularly for initial examination.
Spinning disk confocal/TIRF microscopy specialized for short-term high-speed observation of live cells, particularly of the cell-substrate interface region.
Widefield epifluorescence microscopy for longer-term observation (minutes to hours) of multiple regions of interest in living samples.