Hooker Imaging Core

The News from the Hooker Imaging Core   

LOCATION:  2ND FLOOR TAYLOR HALL

OPEN HOUSE!

The Hooker Imaging Core (HIC) will have an open house in October 6 from 2 PM to 5 PM. Come get to know our instruments and the services we can provide.

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SUPER-RESOLUTION!

Our flagship confocal microscope, the Zeiss 880, has been upgraded to super-resolution with the Airyscan sensor. Those users that need the extra resolution and extra sensitivity are quite pleased with the images.

MAKING A RESERVATION

iLab, our new booking and billing software, is up and running. In order to book time your PI and you must both have iLab accounts. If you are using the Flow Cytometry Core Facility you probably already have an iLab account. If not send Jerry or Robert the names and UNC email addresses of the PI and lab members who will use the HIC including the chart field strings to charge for each person. 

TO BOOK TIME in the HIC via iLab click this link:
https://my.ilabsolutions.com/service_center/3517/ .


FOR TRAINING send email to Robert or Jerry. 


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

    Tier 1 confocal microscopes (there are 3) are $32.08 per hour. 
    Tier 2 confocal microscopes (there is 1) are $20.80 per hour. 
    Tier 3 widefield microscopes (there are 7) are $17.48 per hour.

    Transmission electron microscope is $35.00 per hour.

Image 4  Image 2

The Faculty Advisor is Dr. Jim Bear.

Contact the Directors with questions or interim needs:

            Robert Currin, email: robert_currin@med.unc.edu

            Dr. Jerry Gordon, email: gordon@med.unc.edu 


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.

* 32 channel GaAsP spectral detector (45% quantum efficiency)

* Airyscan detector:

          Greater sensitivity and 1.4 times better resolution than regular confocal

* Autofocus to maintain focus during long experiments

* High speed imaging, 13 fps at 512x512 with zoom > 1

* Live cell imaging with temperature, humidity, and CO2 control

* Tiling and arbitrary multi-position acquisitions

* Lasers: 405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 633 nm

* Wide-field fluorescence: DAPI, GFP, Cy3

* Objectives:

          Plan-Apo 10x/0.45 WD 2.0mm                  

          Plan-Apo 20x/0.8 WD 0.55mm

          Plan-Neofluar 40x/1.3 oil WD 0.21  

          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/0.25

          PlnN 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.

* 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.

* Imaging software MetaMorph

* 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)

The Tier 2 Zeiss LSM 510 is updated with the user friendly Zen software. The system is suitable for FRAP and FRET experiments, ratio-imaging, tiling/stitching and a variety of time course acquisitions.

* There are two PMT detector channels for confocal imaging

* The third confocal channel is the Meta spectral detector with 32 channels

* Laser lines: 458 nm, 488 nm, 514 nm, 543 nm, 633 nm

* CFP/YFP FRET using 458 nm and 514 nm laser lines.

* Motorized x-y scanning stage

* Objectives:

          Plan NeoFluar 10x/0.3 WD 5.6mm  

          PlanApo 20x/0.75

          C Apochromat 40x/1.2 water WD 0.29mm

          PlanApo 63x/1.4 oil WD 0.18mm

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.

* Now 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

* Software: Metamorph

* 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