Microscopy

The Core offers state-of-the-art imaging technology that provides capabilities for:

  • High quality digital microscopy with a wide range of objectives (1X – 100X): Many IDDRC investigators use the fluorescence and confocal microscopes offered by the Core to assess the effects on neuronal structure over development, in response to alteration of gene expression, or in response to environmental or pharmacological manipulations.
  • Quantitative, high-resolution imaging of molecular architecture and cellular ultrastructure: the Array Tomography System, in combination with ultrathin plastic sectioning of fixed tissue and cells allow for visualization of fluorescently labeled proteins at a higher resolution than standard confocal microscopy. IDDRC investigators have taken advantage of the system to localize molecules of interest relative to the cellular architecture of the developing synapse.
  • Multi-label fluorescence imaging that allows intracellular localization and co-localization of large number of antigens (up to 10): Investigators can visualize fluorescently labeled molecules from ultrathin sections using Array tomography and then elute the antibodies to one set of antigens, and re-stain the section a number of times. This allows for detection of many proteins in the same sample. Moreover, immunofluorescent-labeled sections imaged by Array Tomography can then be processed for scanning electron microscopy for finer structural details.
  • In vivo and live cell imaging over hours to days: The Spinning Disk Confocal microscopy provides capabilities to monitor long-term cellular processes in an environment where temperature, humidity and CO2 levels are controlled.
  • High temporal resolution time-lapse imaging using confocal microscopy: The Spinning Disk Confocal also offers rapid acquisition of images at video rate, allowing for visualization of transient molecular interactions in neurons and glia and for monitoring movement of molecules within cells.
  • Advanced microscopy methods including FRAP, FRET and multiphoton: These features, offered by the LSM 710 and LSM 700, take advantage of fluorescence labels to monitor real-time molecular interactions and changes in the concentration of proteins of interest.
  • Quantitative calcium (as well as other cations such as magnesium and zinc) imaging with the confocal microscope: Using the confocal microscopes offered by the Core, IDDRC investigators can take advantage of calcium- dependent fluorophores, or other ion-dependent fluorophores to monitor real-time changes in cation concentrations in single cells as well as in populations of cells.
  • On-site Consultation: In addition to assistance on experimental design and interpretation, the core offers full-time technical help for all users. Dr. Anthony Hill is available on-site to provide advice, help optimize microscope or software settings for specific experiments, trouble-shoot equipment issues and train users on the use of each instrument or software workstation.

Instrument Information

Olympus FVMPE-RS multiphoton Microscope - Reserve

  • Custom modified Olympus FluoroView microscope base with 4X (0.15NA) dry objective, 20x (1.0 NA) water immersion objective.
  • MaiTai DeepSee tunable IR laser
  • GaAsP NDD
  • Piezo Z focus
  • Resonant scanner

Zeiss LSM710  Laser Scanning Confocal/Multiphoton Microscope - Reserve

  • Zeiss upright microscope base with 10X (0.3NA) dry objective, 20x (0.8NA) dry objective, 40X (1.3NA) oil, 60X (1.2NA) oil, and 100X (1.3NA) oil immersion objectives, in addition to a 63x (0.9 NA) and a 20x (1.0 NA, 5.3mm WD) water immersion lens for live specimen imaging. 
  • Argon laser (454,488/514 nm)
  • He/Ne lasers (594 and 633 nm)
  • LED laser (561nm)
  • MaiTai DeepSee tunable IR laser
  • Spectral detection
  • GaAsP NDD

Zeiss LSM700 Laser Scanning Confocal Microscope - Reserve

  • Zeiss Axio Imager.M2 microscope base with 10X (0.3NA) dry objective, 25x (0.8NA) multi-immersion and 63X (1.4NA) oil objectives.
  • Diode lasers emitting at 405nm, 488nm, 555nm, and 635nm
  • X-cite 120 Fluorescence illumination system
  • Filter sets for imaging blue, green and red fluorophores
  • Fujitsu Siemens Esprimo P7935 workstation running Windows Vista Ultimate with dual core 3.0 GHz processor, 4 GB RAM, 2 456 GB internal hard drives, storage devices and drives: DVD, CD-RW, DVD+R/RW, DVD+R/RW-R/RW,
  • HP LP3065, Flat panel, 30” monitor
  • Installed software: Zeiss ZEN image acquisition software, FRAP module

Zeiss Array Tomography system - Reserve

  • Zeiss Axio Imager .Z2 upright microscope base with 10X (0.45NA) dry, 20x (0.8NA) dry, 40X (1.3NA) oil and 63X (1.4NA) oil, objectives.
  • X-cite 120 Fluorescence illumination system
  • Filter sets for imaging blue, green, red and far red fluorophores
  • Fujitsu Siemens Celsius R650 workstation running Windows XP Professional with 2.6 GHz processor, 4 GB RAM, 1 TB internal hard drive, storage devices and drives: DVD, CD-RW, DVD+R/RW, DVD+R/RW-R/RW,
  • HP LP2275w 22” monitor
  • Installed software: Axiovision Rel 4.8 image acquistion software including modules for multichannel, time, Z, mark and find, automeasure, colocalization and deconvolution

Perkin Elmer Ultraview Vox Spinning Disk Confocal - Reserve

  • Nikon Eclipse Ti inverted microscope with computer-controlled, motorized X-Y stage. Objectives include 10X (0.3NA) dry objective, 20x (0.75), 40X oil objective (1.3NA) and 60X (1.4NA) oil objective. 
  • Diode Lasers 50 mW 405nm, 40mW 445nm, 50mW 488nm, 25mW 514nm, 50mW 561nm, and 40mW 640nm with appropriate filters to allow imaging of blue, cyan, green, yellow, red, and far red fluorophores.
  • Motorized stage (X,Y,Z) for Z stack acquisition, time lapse of multiple locations on slide or plate. Adjusting clamp accomodates are wide range of slides, plates and dishes
  • Enclosure for regulation of temperature and O2/CO2 during live specimen imaging
  • X-Cite 120 Fluorescence illumination system
  • Dell Precision T5400 workstation with quad core 3.16 GHz processor running 64 bit Windows XP Professional, 72 GB RAM, 1 TB internal hard drive, storage devices and drives: DVD, CD-RW, CD-ROM
  • Dell ultrasharp widescreen 30” flat panel monitors 
  • Installed software: Volocity image acquisition software

Olympus DSU Spinning Disk Confocal - Reserve

  • Olympus IX2 microscope base with 20X (0.8NA) dry objective and 63X (1.4NA) oil objectives.
  • Dual HGLGPS Fluorescence illumination systems
  • Interchangeable disks for choosing resolution/speed
  • Filter sets for imaging blue, green and red fluorophores
  • Hammamatsu ImagEM X2 EM-CCD camera

Nikon Ti Eclipse inverted microscope - Reserve

  • Nikon Ti Eclipse microscope body with a full complement of infinity-corrected air and oil objective lenses: 1X (0.04NA), 10X (0.3NA), 20x (0.5NA), and 40x (0.75NA) dry objectives; 20X (0.75NA, multi-immersion), 40X (1.3 NA), 60X (1.4NA), and 100X (1.3NA) oil immersion objectives; the microscope has multi-line epifluorescence (DAPI, GFP, RFP, and Cy5 filter sets), brightfield, phase and differential interference contrast optics
  • Andor Zyla sCMOS camera with 2048x2048 pixels
  • Sola LED illumination system

Leica Fluorescence stereomicroscope - Reserve

  • Leica M80 optics carrier with .5x objective, 10x ergonomic oculars
  • IC80 HD digital amera
  • X-cite 120 Fluorescence illumination system
  • Fluorescence filter turret

Optogenetics culture system - Reserve

  • 2 470nm LEDs for activating channel rhodopsin
  • 2 585nm LEDs for activating halo rhodopsin
  • heatted stage insert with brain slice holder
  • peristaltic pump

Nikon widefield microscope (oil objectives) - Reserve

  • Nikon E800 microscope body with dual 35 mm camera system, and a full complement of infinity-corrected air and oil objective lenses: 1X (0.04NA), 10X (0.3NA), 20x (0.5NA), and 40x (0.75NA) dry objectives; 20X (0.75NA, multi-immersion), 40X (1.3 NA), 60X (1.4NA), and 100X (1.3NA) oil immersion objectives; the microscope has multi-line epifluorescence (DAPI, fluorescein, GFP, rhodamine, Texas red, Cy3, and Cy5 filter sets), brightfield, darkfield, phase and differential interference contrast optics
  • Spot RT cooled CCD camera (color & b/w imaging)
  • X-Cite 120 Fluorescence illumination system
  • Dell OptiPlex GX270T 3.20 GHz equipped with 1.0 GB RAM, 120 GB internal hard drive, 8x DVD+RW and 16x CD-ROM, Zip 250 disk drive, 3.5 inch, 1.44 MB floppy drive, 128 MB Nvidia GeForce FX 5200 graphics card
  • Dell Ultrasharp 1901FP, Flat panel, 19” monitor
  • Installed software: Windows 2000 Pro, Spot camera drivers, MS Internet Explorer

Nikon widefield microscope (dry objectives) - Reserve

  • Nikon E80i microscope body with dual 35 mm camera system, and a full complement of infinity-corrected dry objective lenses: 1X (0.04NA), 2X (.1NA), 4X (.02NA), 10X (0.3NA), 20x (0.5NA), and 40x (0.75NA) and 60x (.95NA). the microscope has multi-line epifluorescence (DAPI, fluorescein, GFP, rhodamine, Texas red, Cy3, and Cy5 filter sets), brightfield, darkfield, phase and differential interference contrast optics.
  • Hamatsu C10600-10B ORCA-R2 CCD camera (color and b/w imaging)
  • X-Cite 120 Fluorescence illumination system
  • Dell Precision T3400 with a dual core 2.39 GHz processor, 4 GB RAM, 232 GB internal hard drive, 12x DVD+RW and 48x CD-ROM, 128 MB Nvidia Quadro NVS 290 graphics card
  • ProView, Flat panel, 17” monitor
  • Installed software: Windows XP 64 bit, HCImage image acquisition software

Metamorph Advanced Image Analysis Workstation - Reserve

  • Dell Precision T3400 dual core 2.39 GHz processor, 4.0 GB RAM, 232 GB internal hard drive, 8x DVD+RW and 16x CD-ROM,128 MB Nvidia Quadro NVS 290 graphics card Running 64 bit Windows XP Professional
  • Dell Ultrasharp 1901FP, Flat panel, 19” monitor
  • Installed software: Image J, Metamorph image analysis software

Volocity Image Analysis Workstation - Reserve

  • Dell Precision T5400 quad core 3.16 GHz workstation equipped with 48.0 GB RAM, 931 GB internal hard drive, 8x DVD+RW and 48x CD-ROM, 1.44 MB floppy drive, 128 MB Nvidia GeForce GTX 280 graphics card. Running 64 bit Windows XP Professional
  • Dell Flat panel, 19.” monitor
  • Installed software: Image J, Volocity image analysis software, Zeiss LSM

Imaris Advanced Image Analysis Workstation - Reserve

  • Dell Precision T3500 quard core 3.07 GHz CPU equipped with 24.0 GB RAM, 460 GB internal hard drive, 16x DVD ROM, NVIDIA Quadro FX 580 graphics card
  • Dell Professional P2010H Widescreen, Flat panel, 20” monitor
  • Installed software: Image J, Imaris image analysis software