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Center for Brain Science - Imaging Core Facility (FAS)

Summary:

CBS hosts an optical imaging facility that houses advanced devices for widespread use by neuroscientists and is developing the next generation of optical techniques. The optical imaging is intended to be an extension of individual labs, housed in shared space. By pooling equipment, highly skilled technical management of the tools is available, allowing for routine technological upgrades, quicker troubleshooting, and expert advice. In addition, the joint space leads to camaraderie amongst users, and allows new technical advances in one lab to spread rapidly to other labs. This core facility also frees up space in individual labs that can be used in other ways. Finally, the shared equipment lowers barriers to adoption of the latest technology.

This core facility provides:

* Laser scanning microscopes with motorized stages for high throughput reconstructions of the nervous system
* A histology suite for brain sectioning and tissue preparation
* Stereo fluorescence macroscopes
* The newest tools for high-resolution optical microscopy
* Ultra-fast optical scanning microscopes

Affiliations:

People:

    Resources:

    Instruments

    • Nikon N-STORM super resolution microscope (Ti-E TIRF with Perfect Focus System) ( Total internal reflection fluorescence instrument )

      "The Nikon N-STORM is a super-resolution digital microscope system that integrates 'Stochastic Optical Reconstruction Microscopy' technology. It enables imaging at significantly higher resolution than is possible with conventional light microscopes. The lateral (XY) resolution is ~20-30nm, and the axial (Z) resolution is ~50-75nm.

      This microscope system has TIRF optics and an inverted microscope stand. It has 405nm, 458nm, 561nm and 647nm laser lines. The microscope is fully motorized and has an EM-CCD camera (Andor iXon Ultra DU897). Images are acquired using NIS-Elements.

      The microscope has 40x, 60x and 100x oil immersion objectives. It also has a 10x air objective."

    • Nikon TIRF microscope ( Total internal reflection fluorescence instrument )

      "This microscope system has TIRF optics and an inverted microscope stand. It is the only inverted microscope in the imaging facility. The system is primarily used for automated time lapse imaging of cell cultures. The microscope is fully motorized and has a Hamamatsu CCD camera. Images are acquired using IPLab or NIS-Elements.

      There are filter cubes for DAPI, GFP, TRITC and Texas Red. The system also has an argon-ion laser that is used for TIRF imaging.

      The microscope has oil and water immersion objectives. It also has 4x and 20x air objectives.

      The system is equipped with an incubation chamber that covers the top half of the microscope. A biological gas mixture (5% CO2) can be bubbled into the chamber, and an Air-Therm heater circulates and warms the air in the chamber."

    • Olympus FV1000 confocal microscope (FVA) ( Confocal microscope )

      "This system has 440nm, 458nm, 488nm, 514nm, 561nm and 633nm laser lines.

      The filters are suitable for imaging CFP and YFP and far red fluorophores. There are two spectral channels, two filter channels and a transmitted light channel.

      The system is mostly used to image slides and has the multiple area time lapse (MATL) software module to allow automatic acquisition of tiled image stacks.

      The microscope has a set of oil immersion objectives and a 10x air objective."

    • Olympus FV1000MPE confocal microscope (FVB) ( Two-photon confocal microscope )

      "This system has 405nm, 440nm, 458nm, 488nm, 514nm, 561nm and 633nm laser lines. The 405nm laser is on a SIM scanner separate from the primary scanner and is used for photoconversion and uncaging. The system also has a multiphoton laser (MaTai BB).

      The filters are appropriate for imaging CFP and YFP. The system has four filter channels. There are two external channels for multiphoton imaging.

      The system is used mostly to image slides and has the MATL software module to allow acquisition of tiled image stacks.

      The system has a set of oil immersion objectives and a 10x air objective.

      The microscope has ample working distance for live imaging. There are two low magnification high NA dipping cone objectives (20x 0.95NA and 25x 1.05NA) that are useful for multiphoton and live imaging."

    • Video-rate multiphoton microscope ( Laser scanning confocal microscope )

      "Note: The system requires special training to operate.

      This multiphoton microscope system is custom built using parts from an upright Olympus stand. A resonant galvo is used to achieve fast scan rates (15 to 30 frames per second). The multiphoton laser comprises a Coherent Mira 900 Ti:S oscillator and a 18W Verdi solid-state pump laser.

      There are two external detectors with dichroic filters for R/G or CFP/YFP."

    • Zeiss fluorescence microscope with ApoTome attachment ( Fluorescence microscope )

      "This wide-field microscope has filter cubes for DAPI, CFP, GFP, YFP, Texas Red and CY5. The condenser is set up for dark-field and DIC optics.

      The microscope has an Axiocam MRm (monochrome CCD) camera and the AxioVision software. The stage is not motorized, so it is not possible to acquire montage images automatically.

      The ApoTome structured-illumination attachment allows optical sectioning using the CCD camera. The AxioVision software allows automatic collection of multi-channel Z-stack images.

      The microscope has a set of oil immersion objectives and a 10x water immersion objective."

    • Zeiss LSM-510 upright confocal microscope ( Confocal microscope )

      "This system has 458nm, 488nm, 514nm, 543nm and 633nm laser lines. There is no 440nm line for imaging CFP.

      The filter sets are useful for imaging FITC, TRITC and far red fluorophores (eg, cy5 or alexa-647). The system is not particularly useful for imaging CFP and YFP. There are two fluorescence channels, a transmitted light detector and a Meta detector.

      The Meta detector allows sampling of the emission spectrum (simultaneously acquiring up to 8 channels at a time). Linear unmixing can be used to separate fluorophores with overlapping emission spectra (eg, GFP and YFP).

      The condenser has been removed to allow ample working space for live imaging. The system has dipping cone objectives as well as oil and water immersion objectives."

    • Zeiss LSM-710 upright confocal microscope ( Confocal microscope )

      "This microscope system has 440nm, 458nm, 488nm, 514nm, 561nm, 594nm and 633nm laser lines.

      The primary dichroics are suitable for simultaneous excitation of CFP and YFP. The system will have two spectral channels and a 32-channel quasar detector (for lambda scans).

      The stage is motorized to allow automated tiled image acquisition."

    • Zeiss Pascal confocal microscope ( Confocal microscope )

      "This system has 436nm, 458nm, 488nm, 514nm and 543nm laser lines. There is no 633nm line for imaging far red fluorophores such as cy5 or alexa-647.

      The filters are appropriate for imaging CFP and YFP. The confocal system has two fluorescence channels and a transmitted light detector.

      The LSM software has the Multi-Time macro to allow automatic acquisition of tiled image stacks. The microscope also has an Axiocam HRc (color CCD) camera and runs the AxioVision software to allow automatic tiled multi-channel image acquisition. The microscope has filter cubes for DAPI, CFP, FITC, YFP, Texas Red and Cy5. The condenser is set up for dark-field and DIC imaging.

      The microscope has a set of oil immersion objectives and a 10x water immersion objective."

    Services

    • Microscopy instrument access ( Access service )

      "The microscope systems in the imaging facility are made freely available to members of CBS , MCB and other departments."


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    Last updated: 2015-01-16T10:06:12.829-06:00

    Copyright © 2016 by the President and Fellows of Harvard College
    The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016