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Hooker Imaging Core

The News from the Hooker Imaging Core

LOCATION: 2ND FLOOR TAYLOR HALL

Zeiss 880 with Airyscan has been upgraded with Fast Airyscan for high-speed confocal with increased resolution.
Airyscan gives a 1.7x improved resolution in x, y and z and improves the signal to noise ratio 4-8x. Fast Airyscan is 4x faster than Airyscan or confocal with some loss in resolution but higher than confocal.

Epithelium-Fiber Cell Transition In Galago Lens
GalWGA7-2-18-6a. Overview of Galago epithelium at the fulcrum near the equatorial plane. Vibratome section (120 µm thick) stained with Wheat Germ Agglutinin (WGA) for cell surface (red, 488 nm laser), Phallodin-F-Actin-Alxexa 561 (green, 567 nm laser), DAPI for nuclei (blue, 405 nm laser). Portions of four Radial Cell Columns (RCC1-4) are visible. Nuclei from the epithelium, early fiber cells and a portion of the bow zone are visible. Conclusion: All key features of the outer lens surface at the transition from epithelium to fiber cells is very well preserved and delineated. Filamentous actin is concentrated at the Epithelium- Fiber Cell Interface and along the irregular interfaces between adjacent RCCs.

 

MAKING A RESERVATION

The HIC uses iLab for making equipment reservations and billing. If you have already created an iLab account for another core on campus you do not need to create another one. Follow the link below to create a new account or make a reservation.

https://uncch.ilab.agilent.com/account/login

SUMMER 2021 TRAINING The HIC is transitioning to new Director in Fall 2021. For summer training inquiries, please contact Interim Director Paul Risteff, pristeff@email.unc.edu

PRICING by instrument is on the iLab site. For quick reference our prices for internal unassisted use are as follows.

Tier 1 confocal microscopes (there are 3) are $31.15 per hour.
Tier 2 confocal microscopes (there is 1) are $16.40 per hour.
Tier 3 widefield microscopes (there are 7) are $14.40 per hour.

VivaView is $10 per hour.

Transmission Electron Microscope is $50.50 per hour for independent users.

EM Faculty Advisor: Dr. Joe Costello, joe_costello@med.unc.edu

LM Faculty Advisor:  Dr. Stephanie Gupton, stephanie_gupton@med.unc.edu

EM Research Specialist and Interim Director: Paul Risteff, pristeff@email.unc.edu

spinning beads gupton

Brief specifications of selected equipment

The Tier 1 Zeiss 880 confocal laser scanning microscope offers high sensitivity, (enhanced resolution in x, y and z,) and high image-acquisition speed.

  • Mounted on an inverted Axioobserver Z1
  • Live cell stage top incubator
  • Definite Focus to prevent focal drift
  • 34 Channel GaAsP detector and transmitted light detector (DIC)
  • Lasers: 405nm, 458nm, 488nm, 561nm, and 633nm
  • Objectives: Plan-Apo 10x/0.45 WD2.0mm, Plan-Apo 20x/0.8WD 0.55mm, Plan-Neofluar 40x/1.3 oil WD0.21, Plan-Apo 63x/1.4 oil (partial list)

The Tier 1 Olympus FluoView FV1000 is designed for high-resolution, confocal observation of both fixed and living cells. In addition, the FV1000 offers a revolutionary synchronized laser scanning system called the SIM Scanner. While one laser stimulates, the second laser simultaneously provides high-resolution imaging. This coordination of laser stimulation and imaging makes the FV1000 an ideal choice for FRAP, FLIP and photoactivation with simultaneous observation

  • Environmental chamber for maintaining living cells
  • Tiling and imaging multiple locations
  • 4 confocal detectors: two are spectral and two use bandpass filters
  • Lasers lines: 405 nm, 440 nm, 488 nm, 515 nm, 559 nm, 635 nm
  • CFP/YFP FRET
  • Wide-field fluorescence: DAPI, GFP, Cy3
  • Objectives: UPlanSApo 4x/0.16 WD 13mm, UPlanSApo 10x/0.4 WD3.1mm, UPlanSApo 20x/0.75 WD 0.55mm, PlanApo 60x/1.42 oil WD 0.150mm, LUMPlanF IN 60x/1.00 water WD 2mm, UPlanSApo 30x/1.05 Silicon WD 0.880mm

Our newest confocal microscope. This Tier 1 upright scope will be useful for cleared tissue, zebrafish, any wet preps with dipping lenses, and general fixed specimens

  • Laser lines: 405 nm, 488 nm, 561 nm 633 nm)
  • 3 GaAsP confocal detectors. The emission path uses variable dichroic mirrors to achieve flexibility of bandpass adjustment similar to spectral detectors that use a prism or a diffraction grating.
  • Tiling is easy to set up, very user friendly.
  • Wide-field fluorescence: DAPI, GFP, Rhod
  • Objectives: Fluar 5x/0.25PlnN 10x/0.3 DIC, Pln Apo 20x/0.8 DIC, LD LCI Pln Apo 25x/0.8 oil, EC PlnN 40x/1.3 oil DIC, Pln Apo 63x/1.4 oil DIC

The VivaView Incubator Fluorescence Microscope provides a fully integrated and motorized inverted microscope to allow high quality, long-term time-lapse imaging in a constant and optimized environment. Multiple locations in each of up to eight 35 mm dishes can be imaged for each time point with fluorescence and/or DIC. VivaView is $10 per hour

  • Objective: UPLSAPO20x (NA 0.75, WD 0.6mm)
  • Intermediate magnifications (zoom factors) 0.5x/1x/2x, Motorized magnification changer for final magnifications of 10X, 20X, 40X
  • Morphological observation (Transmitted light) Light source 1W LED (625nm) Contrast method Differential Interference Contrast (DIC) Condenser NA 0.55, FS fixed (rectangle), DIC prism fixed
  • Fluorescence Observation (incident light) Light Source X-Cite eXacte Pre-aligned 2000 hour DC lamp with Closed – Loop Feedback, Adjustable Iris, Liquid Light Guide Excitation change Motorized 6 position filter wheel, 1 for transmitted light, 5 for fluorescence
  • Fluorescent Filters: GFP, RFP, CFP, YFP
  • Motorized Stage Stroke 10mm x10mm, each dish Sample Capacity Glass bottom dishes with 35mm diameter, up to 8 Sample Tray Can be easily put on and taken off, autoclave capability, include 8 lids with glass window for the glass bottom dishes with 35mm diameter. Mat-Tek dishes recommended
  • Focus Objective drive, motorized up/down control Operation Jog dial control
  • Camera: Hamamatsu Orca R2 cooled CCD camera, -35oC air cooled 2/3″ (8.67mm x 6.60mm), 1344 x 1024 pixels 12 bits or 16 bits
  • Liquid handling assist Motorized open/close control of designated dish. Manual pipetting through the small opening on the internal door of the incubator
  • MetaMorph Imaging Software
  • CO2 Incubator Temperature 37°C Accuracy +/- 0.3°C (ambient temperature 25oC, no load) Humidity 95% +/- 5% RH CO2Concentration 0-20% Accuracy +/- 0.15% (ambient temperature 25°C, no load)

Fluorescence microscopy for automated high content screening (cells, organelles, tissues, organs)* Can read plates up to 1536 wells in addition to slides, with environmental control & liquid handling:

  • Fast-image a one color 1536 well plate in under 20 minutes
  • Solid-state illumination source, sCMOS camera
  • Five fluorescent filter sets & multiple objectives
  • Hardware and Software autofocus

The Tier 3 Nikon 2000E inverted widefield micorscope is a high performance system that can perform complex and rapid acquisition tasks. It is one of several widefield systems in the HIC. The system is equipped with a Hamamatsu Flash 4 CMOS camera for rapid and sensitive detection of fluorescent and transmitted light signals. It is also equipped with an encoded stage and filter wheels for time lapse imaging, tiling/stitching and multi-point acquisition among others. Image acquisition and microscope devices are controlled through MetaMorph.

  • Equipped with 4 color Sedat filter set to speed multi-color acquisitions and and allow the eye and camera to see the same signal
  • DAPI, FITC/GFP, Texas Red, Cy5
  • Camera: ORCA-Flash4 sCMOS, (2048 x 2048 pixels, 6.5µm pixel size, 13.3mm x 13.3mm chip area)
  • Light Source: Sola SE II solid state illuminator, wavelength range 350-680nm
  • MetaMorph Imaging Software
  • Objectives:
    2x/0.1    10x/0.3, pH1 dl 1.2mm, WD 15.2mm
    40x/0.6 ELWD-PlanFluor, DIC-M, Ph2 WD 3.7-2.7mm
    40x/1.3 PlanFluor, DIC WD0.2
    100x/1.4NA Plan Apo DIC (partial list)

The Leica MZ16FA is a fully automated planapochromatic fluorescence stereo microscope. The filter changer, zoom, focus, UV shutter and double iris aperture are computer controlled which allows multifluorescence experiments to be repeated quickly and accurately.

  • Cameras:
    ORCA ER, (1344(H) x 1024(V) pixels, 6.5µm pixel size, 8.67mm x 6.6mm chip area(2/3-inch format), 8.8 frames/sec
    QImaging MicroPublisher 5.0, (2560 x 1920) pixels, 3.4µm pixel size, 10 bit, color camera
  • Basic Magnification range 7.1x to 115X with 10x eyepiece
  • Motorized zoom (16:1)
  • Automatic control of Multifluorescence Experiments
  • Magnification:
    2x PlanApo (230) Resolution: 840 LP/mm=0.6 µm
    1x PlanApo (115) Resolution: 420 LP/mm

Modern electron optics produce high resolution images of a variety of sample types including thin sections, negative stain on grids and freeze-fracture replicas.:

  • *NEW* Gatan Rio 16 CMOS Camera
    • High sensitivity: Newly optimized scintillator and 1:1 fiber optic coupling. Gpixel CMOS sensor with 100% duty cycle and ultralow readout noise.• High resolution: 9 μm pixel, optimal for 30 – 200 kV operation• Large field of view: Up to 16 megapixels (4k x 4k)• High speed: Up to 20 4k x 4k resolution frames per second and 160 1k x 1k frames per second

    • Easy to use: Gatan Microscopy Suite® technique panels for simplified experimental interaction

    • Flexible: Use Rio across many applications, including routine EM, HREM, in-situ, diffraction, 4D STEM, and more

    • All Rio cameras come standard with drift correction and full resolution image display at live frame rates

    • Automatically compensate for column, holder, or specimen drift when recording images

    • Live, full resolution display: Eliminates yesterday’s need for separate View and Acquire modes, and the full resolution information can be displayed in Fast-Fourier transforms in real time

  • Accelerating voltages from 40kV to 120kV using a tungsten filament; a highly stable goniometer controls x, y, z and tilt positions.
  • Accessories include a Gatan 626 cryostage, pumping station and temperature control unit for examining thin samples embedded in amorphous ice using low-dose conditions.
  • Sample preparation equipment available for cutting thin sections, freezing thin samples on grids and freezing larger samples with a high pressure freezer.
  • Assistance and training available for sample preparation and examination of specimens.