Hooker Imaging Core
The Hooker Imaging Core (HIC) is an open core providing instrumentation, training and expertise in light and electron microscopy for the UNC biomedical research community and outside users.
LOCATION: 2ND FLOOR TAYLOR HALL
ACCESS
The HIC is open for use by researchers from UNC-CH and beyond.
The HIC uses iLab for making training requests, 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, make a reservation or request training on a light microscope.
https://uncch.ilab.agilent.com/account/login
Acknowledgement of the HIC is required in all publications that include data from HIC resources. Here is a template: Microscopy was performed at the UNC Hooker Imaging Core Facility, supported in part by P30 CA016086 Cancer Center Core Support Grant to the UNC Lineberger Comprehensive Cancer Center.
PRICING
Pricing by instrument for internal and external customers is listed on the iLab site.
For quick reference our prices for internal unassisted use are as follows.
Tier 1 confocal (there are 2) | $34.40/hour |
Tier 2 confocal (there is 1) | $19.20/hour |
Tier 3 confocal (there are 7) | $18.80/hour |
VivaView | $10.50/hour |
Transmission Electron Microscope | $50.50/hour |
Training and advanced assistance | $40/hour |
LEADERSHIP
Director: Wendy Salmon, wendy_salmon@med.unc.edu 236 Taylor Hall
EM Research Specialist: Paul Risteff, pristeff@email.unc.edu
EM Faculty Advisor: Dr. Joe Costello, joe_costello@med.unc.edu
LM Faculty Advisor: Dr. Stephanie Gupton, stephanie_gupton@med.unc.edu
Brief specifications of selected equipment
- 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)
- 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 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
- 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
- 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)
- 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/0.110x/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)
- 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
- *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.
NEWS
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.