Integrated Cellular Imaging
Emory University
Contact: Laura Fox-Goharioon, Core Director
The Emory Integrated Cellular Imaging (ICI) core provides state-of-the-art light microscopy and image analysis platforms. ICI offers confocal, spinning disk confocal and live cell imaging, multi-photon animal and tissue imaging, widefield with deconvolution, super resolution, light sheet and image analysis. To effectively implement these technologies, ICI provides consultations, expert training, and support for all our systems. We assist our users from bench to publication, starting with designing experiments tailored to the specific microscope, through training and data acquisition, to data analysis and interpretation.
If you use the ICI for your research, please acknowledge the ICI in your publications and include its RRID (RRID:SCR_023534).
Confocal Imaging
Leica SP8 (Winship): The Leica SP8 confocal laser scanning unit, attached to the latest Leica inverted DMi8 body, comprises an infinitely flexible prism based spectral detector system and multiple excitation laser lines (405, 488, 514, 561 and 633nm) for a vast variety of experimental and labeling options (coupled with LASX’s unmixing software for experiments with potentially 10 or more colors). Four channel detection, via two PMT’s and two extremely sensitive Leica HyD detectors (GaAsP based), together with resonant scanning provides high speed imaging. A transmitted light detector allows for simultaneous laser scanned DIC or phase imaging. A stage and objective warmer facilitates numerous live cell imaging approaches such as FRET, FRAP, and photoconversion. High speed automated tiling/stitching and multipoint location acquisitions are also possible in combination with the above. Software options include FRAP XT, FRET AB and SE (acceptor photobleaching and sensitized emission) as well as Navigator software for tiling.
Olympus FV1000 (Whitehead): This FV1000 confocal laser scanning microscope is mounted on an inverted Olympus IX81 microscope with a motorized x-y-z stage. It has a three channel PMT detection system with its filter based detection on all channels. In addition, a transmitted light detector provides laser scanned DIC for label free imaging. These coupled with multiple laser lines (405, 488, 515, 543, 635) to facilitate highly flexible imaging combinations.
Live cell confocal imaging
Nikon A1R HD25 (Whitehead): This live cell confocal laser scanning microscope is optimized for imaging cellular dynamics at high speed and a large view of view (25mm). A hybrid dual scanning confocal system with both galvanometric and resonant scanners, together with a piezo z controller, can acquire 1024x1024 images at 15 fps, 512x512 images at 30 fps, or with a reduced size up to 720 fps. A stage top environmental chamber at 37° C and 5% CO2 required for live cell imaging, coupled with a Perfect Focus system, provides extremely stable long-term imaging without drift. Flexible laser scanning control for photokinetic experiments can be conducted in multiple user-defined regions of interest. Six laser lines (405, 440, 488, 514, 561, & 640nm) and four channel PMT (2x GaAsP, 2x multi-alkali) detectors, in addition to a transmitted light channel for laser scanned DIC provide flexible live cell experimental options. A DUVB-2 GaAsP spectral detector unit provides arbitrary 10 – 320 nm wide spectral windows between 400 and 720 nm for complete spectral flexibility. NIS Elements packaged with analysis options coupled to JOBS tailorable data-based control system.
Nikon Ti2 – Crest X-Light Spinning Disc Confocal (Winship): A Nikon controlled Ti2 with a Crest Optics X-Light large FOV ultrafast spinning disk confocal provides a 18mm FOV utilizing a sensitive high speed sCMOS camera (~100fps or more for cropped ROI). A stage top environmental chamber at 37° C and 5% CO2 required for live cell imaging, coupled with a Perfect Focus system, provides extremely stable long-term imaging without drift. A fast z-piezo and motorized xy stage provides high speed automated multi-location and tiling/stitching acquisitions. Seven laser lines (408, 445, 473, 518, 545, 635, & 750nm), DIC, and phase acquisitions are available. NIS Elements packaged with analysis options coupled to JOBS tailorable data-based control system.
Nikon SoRa Spinning Disc Confocal (HSRB I): The CSU-W1 SoRa is a confocal field scanning spinning disk system and a confocal super resolution system in a single package. We offer dry as well as water immersion lenses for high resolution imaging with fixed and live cell. The Nikon SoRa include two spinning disks which can be exchanged easily via software control: a super resolution disk which also includes the capability of optical sectioning, and a standard confocal imaging disk. The confocal offers spinning disk confocal, brightfield, widefield and allows for time-lapse imaging and tiling. The system is equipped with four laser lines, air, water immersion, and oil immersion objectives, an environmental chamber for live cell: Tokai Hit STXF, and universal sample holders. The microscope body is a motorized inverted Nikon Eclipse Ti2-E with hardware triggering for multicolor imaging without switching emission filters. NIS Elements packaged with analysis options coupled to JOBS tailorable data-based control system
Leica Stellaris 8 (HSRB II): A point scanning system, including a White Light Laser as excitation light source. An extended detection range up to 850 nm plus the expanded excitation range in the visible from 440 nm up to 790 nm allow the application and separation of an extended range of spectrally overlapping fluorophores, up to five simultaneously. Point scanning inverted confocal equipped with Galvano and resonant scanners, 5 detectors: 3 Power HyD-S and 2 Power HyD-X, a Tokai Hit stage top incubator for CO2 and temperature control for live cell imaging. The system has White light lasers, ranging from 440-790nm as well as a 405-diode laser for DAPI. The Stellaris includes Navigator software for tiling , TauSense for FLIM, and LIGHTNING for optimal extraction of image details and maximum resolution (Deconvolution).
Super-resolution and Light Sheet imaging
Abberior STED Super Resolution Microscope (Whitehead): The Facility Line STED microscope from Abberior Instruments has four lasers for excitation (405, 485, 561, 640 nm) as well as two pulsed STED lasers (595 and 775 nm). Patented adaptive illumination methods enable live-cell imaging with drastically reduced photobleaching and phototoxicity. FLIM capabilities available through two channel TCSPC. The microscope is equipped with Z-drift compensation unit, an Okolab stage top incubator for CO2 and temperature control (Uno-T line) and is capable of multi-channel, z-stacks, time-lapse imaging, multi-point capture, tiling as well as FLIM, FRAP, FLIP, FRET, photoactivation, and colocalization.
Nikon N-SIM (Whitehead): The Nikon Structured Illumination Microscope (SIM) doubles the resolution of widefield microscopy in all three dimensions (down to 120 nm x-y and 300 nm z resolution) and can image samples prepared with conventional techniques (same as confocal or widefield). A sensitive iXon EMCCD camera and three laser line system (488, 514, and 561nm) provides high speed multi-color acquisitions. A SIM-TIRF mode is also available. The system also includes a 37°C 5% CO2 and humidity-controlled stage insert, together with heated objectives for the option of live cell super resolution acquisitions.
3i Lattice Light Sheet (Clinic B): The Intelligent Imaging Innovations (3i) lattice light sheet is a transformative new technology for live cell imaging with unprecedented spatiotemporal resolution. It combines custom water dipping objectives and structured illumination to provide a resolution 3-4x better than traditional confocal microscopy in SIM mode, or fast volumetric imaging with ~100 times less phototoxicity/photobleaching than confocal in differed mode. The system has four-laser options (405, 488, 561, and 642nm) operating in sequential mode, but without spectral separation for speed optimization. Samples are loaded onto 5mm coverslips and immersed in an 8ml bath below the inclined objective lenses.
Miltenyi Ultramicroscope Blaze LFOV Light sheet microscope (HSRB II): A fully automated light sheet microscope for imaging large or multiple cleared samples at subcellular resolution. The microscope is equipped with 5 laser lines (405, 488, 561, 640, 785) and uses Uni- and bi-directional illumination with 1-6 light sheets. The system houses three objectives, 1.1X, 4X, and 12X, allowing for magnifications from 0.6X up to 30X. iDISCO clearing is the only clearing method that is allowed on the system.
Multiphoton
Leica SP8 (Whitehead): The Leica SP8 is an upright confocal and multiphoton microscope with a Coherent Chameleon Vision II laser tunable from 680 to 1080nm. Two highly sensitive non-descanned GaAsP based HyD detectors provide imaging of multiphoton processes. The confocal unit houses an infinitely flexible prism based spectral detector system and multiple excitation laser lines (458, 476, 488, 496, 514, 561, 594nm) for a vast variety of experimental and labeling options (coupled with LASX unmixing software for experiments with potentially 10 or more colors). Four channel detection, via two PMT’s and two extremely sensitive Leica HyD detectors (GaAsP based), together with resonant scanning provides high speed imaging. A CCD camera allows for brightfield and Dodt Gradient Contrast (DGC) for label free imaging and sample navigation. A stage and objective warmer facilitates numerous live cell imaging approaches such as FRET, FRAP, and photoconversion. A resonant imaging mode provides high speed scanning at up to 428 fps and there is automated tiling and image stitching. Oil, air and water immersion objective lenses are available. Software options include Navigator, FRAP, FRET.
Widefield fluorescence
Leica Thunder widefield microscope( HSRB II): 3D live cell wide field microscope. The Thunder imager helps to maintain optimal physiological conditions by minimizing photobleaching, providing high performance imaging. The microscope is equipped with Motorized DMi8 microscope, fully automated for bulk plate imaging, highly sensitive K8 camera, multi-line, high-intensity fluorescence LED light source, and 750 near IR imaging. The microscope can also do large scale screens as well as live cell imaging with a Tokai Hit stage top incubator.
Keyence BZ-X810 (Winship): The Keyence BZ-X810 automated inverted microscope uses a variety of dry objectives for fluorescence, phase contrast, bright field, and color bright field microscopy. The microscope offers Z-stacks as well as tiling. It has an environmental top cover for light-tight imaging.
Zeiss Axioscan 7 upright bulk slide scanner (HSRB II): The Zeiss Axioscan 7 is a fully automated microscopy system that allows users to quickly and reliably digitize bulk samples. With its automated application flexibility and 7 wavelength Colibri LED light source, the Axioscan 7 can provide high-quality images with minimal effort. The system can scan up to 12 slides in a single pass (expandable up to 100 slides), offers fluorescence, brightfield, polarization, and Transport of Intensity Equation (TIE) brightfield contrast imaging modalities, 7 wavelength Colibri LED light source (385, 430, 475, 511, 555, 590, 630, 735nm), an Automated Application Flexibility – with its fast filter wheels, color, and monochrome cameras, you can maximize multiplex imaging with minimal effort. This system is also capable of unattended, continuous 24/7 operation.
Zeiss Axioplan 2 (Winship): The motorized Zeiss Axioplan 2 is equipped for routine widefield epifluorescence and transmitted light applications (DIC, phase, and brightfield) using a range of high-end Zeiss Plan-Apo objectives. A Zeiss Axiocam color camera allows for histology imaging or fluorescence imaging via multiple filter sets.
Olympus IX51 (Winship): The inverted IX51 can perform epifluorescence and transmitted light microscopy. It contains long working distance objectives that allow for visualization through plastic culture dishes or glass. An Infinity CCD monochrome camera allows for sensitive acquisitions from multiple filter set options.
BioTek Lionheart FX (Whitehead): The compact Lionheart FX widefield microscope provides flexible automated acquisitions in fluorescence, brightfield, and color imaging modes, together with multiple lenses and 37°C and 5% CO2 control allowing live cell multiple plate reading, to IHC, and widefield-deconvolution.
BioTek Lionheart FX (HSRB I): The BioTek Lionheart LX automated microscope offers a variety of dry objectives along with fluorescence, phase contrast, bright field, and color bright field modalities for imaging slides, dishes, and 6 - 1536 well plates. It offers the same imaging and software as found in Whitehead just without the live cell capabilities.
Image Analysis (Across all locations):
Six image analysis workstations are available, ranging from Dell workstations to custom built towers designed to handle 500GB sized data sets. Users can analyze their data using a full suite of image analysis options that have recently been updated. These include Bitplane Imaris, Volocity 6.2 and 3-D analyses. 2D analysis can be performed with Nikon Elements, Fiji Is Just ImageJ (FIJI; open source), CellProfiler (open source), and Zeiss Axiovision. Deconvolution is available via Huygens Essentials. Custom Fiji macros and java plugins can be design and tailored to individual needs, and are available to download from our resources page, together with YouTube analysis tutorials. MATLAB code can be design for stand-alone usage, or for direct interface within Imaris.
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