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The Children's National Research Institute
Light Microscopy and Image Analysis Core
The Children’s National Light Microscopy and Image Analysis Core (CLIC) is located at the Children’s National Research Institute. The core supports research aimed at studying the cellular and molecular processes through the use of optical imaging.
The core offers access to:
- Advanced microscopes
- Hands-on training
- Help with the experimental design
- Technical support to optimally utilize the microscopes
The core has an open-door policy and offers users 24/7 access to the available equipment.
Request core use and reserve the equipment you are trained on.
Manager: Shivaprasad Bhuvanendran, M.Eng.
Director: Jyoti K. Jaiswal, Ph.D.
Olympus FV1000 point scanning confocal microscope for high resolution 3D and multi-color imaging of cells and thick tissue specimens. It is equipped with visible light (405, 442, 488, 515 and 647 nm) and pulsed infrared (Spectra-Physics MaiTai HP-OL) lasers. It Is equipped with one regular PMT, two spectral PMTs, two GaAsP detectors, two non-descanned detectors and a motorized stage. This microscope allows multi-area time-lapse imaging, tiled acquisition of optically sectioned images, imaging live cells and organotypic slices using two-photon microscopy, simultaneous photo-stimulation with 405nm laser, spectral unmixing and FRET.
Leica TCS SP8 point scanning confocal and super-resolution imaging system allows time-gated STimulated Emission Depletion (gSTED) super-resolution microscopy. It uses an Acoustoptical Beamsplitter (AOBS) which in combination with prism, Leica SP detector and supercontinuum White Light Laser (WLL) allows flexibility of simultaneously imaging multiple fluorophores and enables excitation, and emission fingerprinting (spectral unmixing). Use of Leica HyD detector with WLL enables low light detection. By time gating it enables imaging in high sample background and improved super-resolution with gated STED in X/Y/Z planes. It supports multi-area time-lapse confocal imaging and deconvolution microscopy (HyVolution).
Olympus VS120 Digital Pathology system with motorized stage allows efficient and fully automated focusing and digital scanning of large tissues for pathology and research purposes. In addition to low magnification 2X objective for rapid scanning, it is also equipped with 10X - 60X objectives that are optimized for fluorescent imaging.
The Olympus BX63 is an upright epi-fluorescence microscope. It is equipped with a X-Cite Xylis LED light source, halogen lamp and a DP80 dual-chip camera for both bright field and fluorescence image acquisition. It has a wide range of objectives starting from 1.5x to 100x. It allows automated multicolor imaging from UV to far-red region of the optical spectra.
The Zeiss ApoTome2 is a structured illumination microscope on an inverted Axio Observer stand with a motorized stage. It is equipped with 4-LED Colibri (385nm, 475nm, 555nm, 630nm) and an Axiocam 503 Mono camera. It allows live-cell imaging and optical sectioning of thin tissue sections.
MMI CellCut system on an inverted Olympus stand enables contamination-free isolation of cells and tissues from histological specimen. It can be used in both bright field and fluorescence modes.
Leica THUNDER long-term live cell imaging widefield microscope that uses the approach of “Computational clearing” to remove out-of-focus light in real-time to allow high-speed, high-quality 3D imaging using a camera-based setup. This allows rapid and gentle 3D imaging of multicolor stained live specimens. It also allows phase contrast and DIC imaging. The microscope is equipped with an Okolabs stage top chamber for temperature and CO2 control with stage inserts for multi-well plates, petri dishes and regular slides for imaging samples over days. This makes the system well-suited for imaging live (or fixed) spheroids and organoids.
This live cell imaging microscope is custom-built in collaboration with two companies – Olympus and Intelligent Imaging Innovations (3i). It is equipped with Olympus Cell-TIRF and Yokogawa spinning disc confocal devices. It is equipped with six laser lines and standard objectives as well as high NA objectives for TIRF. It supports frequency domain Fluorescence Lifetime Microscopy (FLIM), targeted laser ablation (for focal injury), multi-area time-lapse and tiled optically sectioned imaging of live cells together with FRAP and spectral FRET. It allows automated switching between widefield, confocal and TIRF modalities to image samples.
The Olympus FVMPE-RS is a multiphoton microscope on an upright stand with a motorized stage that can be manually adjusted in z to accommodate imaging model organisms of varied sizes. This microscope uses a Spectra-Physics MaiTai DeepSee multiphoton laser (tunable from 690nm to 1040nm) and a continuous wave 458nm laser. It is equipped with two non-descanned PMTs, two high sensitivity GaAsP detectors and uses both a galvo and a resonant scanner. With equipment for stage incubation and perfusion of organotypic slices this microscope enables imaging live cells and tissues, multi-area time-lapse and tiled 3D images. It also supports using Second Harmonic Generation (SHG), and two-photon FRET imaging, high speed optogenetics and calcium imaging with simultaneous photo-stimulation.
MBF system on an upright Zeiss stand with ApoTome2 is used for stereology and neuron tracing. It has the StereoInvestigator, Neurolucida and Microlucida software for unbiased quantitative analysis of number, density, size and all other stereological quantities, neuron tracing and 3D reconstruction with whole-slide scanning.
Zeiss SteREO Discovery V8 Dissecting microscope with 3D image perception is available for transmitted, darkfield, morphometric and fluorescence analyses of mouse embryos and tissue slices. This system is equipped with 0.8X and 1.6X objectives and filters for DAPI, FITC, RITC and CY5 excitation/emission. Additionally, Zeiss Stemi 2000 and Stemi SV11 stereo dissecting microscopes are available for coarse dissections and the preparation of organotypic slices.
The core users have access to multiple high-end image analysis workstations and virtual machines for image quantitation, processing, 2D/3D image deconvolution and reconstruction, motion picture editing and preparation of images and videos for publications and presentations by the users. Available software packages include:
- Fiji Image J
- cellSens Dimension (with deconvolution)
- Huygens Professional (for deconvolution)
- Imaris (full package)
- Neurolucida 360
- Leica Application Suite (LAS X)
- Zen lite
- CorelDRAW 2019
- Adobe Photoshop CS6
- Adobe Illustrator
- Prism (for statistical analysis)