The Michael Hooker Microscopy Facility (MHMF) is a research light microscopy facility providing advanced digital light microscopy, image processing and analysis resources for users from the UNC Chapel Hill campus. We offer instrumentation and instruction to enable users to acquire, process and analyze images from a wide variety of sample types.
Equipment Available
New! Olympus FV1000 MPE SIM Laser Scanning Confocal Microscope
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- Laser Spot Scanning Confocal Microscope
- Inverted microscope stand with widefield DIC
- Visible excitation lasers switched/modulated using an AOTF
- Multiphoton excitation laser modulated with an AOM
- Simultaneous transmitted light or DIC imaging while scanning confocally (single or two photon)
- Three standard Photo Multipliers Tube (PMT) light detectors with 2 spectral discriminations
- Motorized x-y stage- tilting, tilting with time lapse
- Environmental chamber for controlled temperature, gas, and humidity
- FRAP/FLIP/FLIM/FRET/PhotoActivation/etc. – SIM scanner
- True 12 bit acquisition- 4096 pixel grey level
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Detection:
- Filtering through glass interference filters
- Spectral separation through diffraction gratings
- Two tunable detection bands (1-100 nm band width) with adjustable spectral windows before the three Photo Multiplier Tubes (PMT)
- Standard XY scanning with zoom and rotation
- Transmitted light and DIC (non confocal)
- FRAP, FRET, photoactivation, Emission Spectra, ratio imaging
- Two Channel non descanned detectors (NDD)
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Objectives:
#1.5air
#1.5air
#1.5oil |
| Mag. |
NA |
Type |
WD |
Corrections |
Cover Slip |
Immersion |
| 10x |
0.4 |
UPlansaPO |
3.1 mm |
| 20x |
0.75 |
UPlanSApo |
550 um |
| 30x |
1.05 |
UPlanSApo |
800 um |
corr |
0.13-0.19 |
silicon oil |
| 60x |
1.42 |
PlanApo |
150 um |
More information and how to get started using the Olympus FV1000
Advanced and Standard Microscopy
- Standard transmitted color photomicrographs
- Fluorescence photomicrographs
- Phase contrast and DIC/Nomarski
- Ratio imaging. e.g. Ca2+ Fura-2
- Time-lapse (fluorescence and transmitted light)
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Additional Confocal Microscopy
- Multichannel fluorescence + DIC overlayed
- Fast X-Z scanning 3D and 4D image acquisition
- Spectral Imaging – dye emission unmixing
- Co-localization & analysis
- Time lapse
- Fluorescence Recovery After Photobleaching (FRAP)
- Fluorescence Resonance Energy Transfer (FRET)
- Deconvolution
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Live Cell Imaging
- 3-color Spinning-disc Confocal (FITC, Texas Red & CY5 like dyes)
- Simultaneous fluorescence and transmitted light (DIC)
- Heated stages, chambers, objective heater
- Temperature and environmental control
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Color Brightfield Imaging (high resolution)
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- Nikon DXM 1200 pixel shift color camera (12 MegaPixels)
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| Scattergram showing no co-localization between the red & green antibody labels, Dr. Karen Loechner. |
Mouse Embryo 3D render, Dr. Jaime Rivera. |
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Stereo Fluorescence Micro/Macroscope
- Fluorescence/standard illumination
- Motorized focus & filter changers
- Automated acquisition – z-series/time lapse/multi channel
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Laser Micro-Dissection System
- Isolation of subcellular and tissue regions
- For DNA/RNA amplification or protein for mass spectroscopy
- Cauterization of vessel in developing organism
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Image Processing and Analysis
- Co-localization analysis
- 3-D Image rendering and quantification
- Particle tracking – 2D & 3D
- Fluorescence quantification
- Densitometry
- Area/volume measurements
- C-Imaging, Metamorph (offline)
- Volocity – Decovolution, 3D rendering, 3D quantification, colocalization, tracking
- Volocity remote access license server
http://microscopy.unc.edu/volocity |
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Airway Epithelium, Dr. Raymond Pickles
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“Brainbow” Dr. Alain Burette, Cell & Devel. Biology
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Other Resources
MHMF Personnel and Contact Information
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Dr. Michael Chua
Cell & Molecular Physiology
7201 Marsico Hall
919-265-9380
microscopy@unc.edu |
