Light Microscopy
The Analytic Imaging Facility (AIF) has a range of light microscopes, including confocal and epifluorescent and brightfield microscopes, available for researchers. AIF also offers automated slide scanners.
Confocal Microscopes
Point scanning and spinning disk confocal microscopes for fluorescent samples that need high resolution.
Attention: The NIH requires that all publications using data from this instrument must list SIG # 1S10OD034397-01 in the Acknowledgements.
Objectives:
- 10X NA=0.4 Air
- 20X NA=0.75 Air
- 40X NA=1.3 Oil
- 63X NA=1.4 Oil
Excitation Lasers:
- 405nm
- White Light Laser (440nm-790nm)
Emission: Adjustable emission windows
Microscope: Inverted DMi8
Image Detection: Point scanning with 5 HyD detectors
Additional Features:
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Reflection-based Adaptive Focus Control
- Photoactivation, FRAP/ FLIP, FRET
- Leica Lightning Deconvolution
Attention: The NIH requires that all publications using data from this instrument must list SIG # 1S10OD023591-01 in the Acknowledgements.
Objectives:
- 10X NA=0.4 Air
- 20X NA=0.75 Air
- 40X NA=1.3 Oil
- 63X NA=1.4 Oil
Excitation Lasers:
- 405nm
- White Light Laser (470nm-670nm)
Additional Lasers:
- 440nm (pulsed) for FLIM
- Argon (458nm, 476nm, 488nm, 496nm, 524nm) for FRAP
Emission: Adjustable emission windows
Microscope: Inverted DMi8
Image Detection: Point scanning with 2 PMT and 3 HyD detectors
Additional Features:
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Fast scanning mode (resonant scanner)
- Reflection-based Adaptive Focus Control
- Photoactivation, FRAP/ FLIP, FRET, FLIM
- Leica Hyvolution Deconvolution
Objectives:
- 10X NA=0.45 Air
- 20x NA=0.75 Air
- 25X NA=1.05 Silicone (by request)
- 40X NA=1.4 Oil
- 60X NA=1.4 Oil
- 100x NA=1.45 Oil
Excitation Lasers:
- 405nm
- 488nm
- 561nm
- 640nm
Microscope: Inverted Nikon ECLIPSE Ti2E
Image Detection: Two Hamamatsu ORCA-FLASH 4.0 sCMOS cameras
Additional Features:
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Reflection-based Perfect Focus
- Optimicroscan Device for Photoactivation, FRAP/ FLIP (405nm or 473nm)
Objectives:
- 4X NA=0.2 Air
- 10X NA=0.45 Air
- 20x NA=0.8 Air
- 40X NA=0.8Air
- 60X NA=1.42 Oil
- 100x NA=1.45 Oil
Excitation Lasers:
- 405nm
- 446nm
- 488nm
- 518nm
- 577nm
- 640nm
- 748nm
Microscope: Inverted Nikon ECLIPSE Ti2E
Image Detection: Two Hamamatsu ORCA-FusionBT SCMOS cameras
Additional Features:
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Reflection-based Perfect Focus
Specialized Microscopes
Microscope with optical resolution beyond the traditional diffraction limit, for fluorescent samples.
Attention: The NIH requires that all publications using data from this instrument must list SIG # 1S10OD18218-1 in the Acknowledgements.
Objectives: 100X NA=1.49 Oil
Excitation Lasers:
- 405nm
- 488nm
- 561nm
- 647nm
Microscope: Inverted Nikon ECLIPSE Ti-E
Image Detection: EMCCD Andor iXon3 DU897
Additional Features:
- TIRF, STORM, SIM
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Reflection-based Perfect Focus
Microscope with pulsed laser for deep tissue imaging/intravital imaging.
Objectives: 25X NA=1.05 water
Excitation Laser: Spectra Physics Mai Tai-DeepSee (690-1040nm)
Fixed Fluorescence Channels:
- Blue/second harmonics
- Green
- Red
- Far Red
Microscope: Inverted Olympus IX81
Image Detection: Point scanning with 4 PMT detectors
Additional Features:
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Photoactivation/Photoconversion
Automated Slide Scanners
Automated scanner for fluorescent and brightfield/histology slides.
The 3DHistech P250 Slide Scanner is an automated, high-capacity slide scanner that provides brightfield/histology and fluorescence scanning of whole slides. It is a drop-off service.
Attention: The NIH requires that all publications using data from this instrument must list SIG #1S10OD026852-01A1 in the Acknowledgements.
Objectives:
- 20X NA=0.8 AIR
- 40X NA=0.95 AIR
Fluorescence Filters:
- 405
- 488
- 555
- 633
- CY7
Image Detection:
- Brightfield/histology: High-speed color CMOS camera
- Fluorescence: Low-noise high-speed CCD camera
Additional Features:
- Extended focus
- Z-stacking
- Brightfield/color imaging for histology
- Image analysis package (Quant Center)
Special Rules for Slides:
- Use hard mounting media only (no nail polish, no wet mount)
- No protruding coverslips
- No protruding labels
- Slides need to be clean and dry: clean the back of the slide with ethanol and scrape the edges of the slide with a razor blade
- Use sharpie for circling sample only on coverglass side
What to write on box for drop off:
- Email address:
- PI:
- Grant:
- Multilayer: Yes / No
- Stain: Brightfield / Fluorescence
- For Fluorescence:
- Indicated fluorescence channels:
- 405 / 488 / 555 / 633 / Cy7
- Indicated brightest slide for each channel on the slide with a * and put it in slot #1 in the box
Instructions to Download Viewer Software:
Slide Viewer:
- Go to: https://www.3dhistech.com/software/slide-viewer/
- Click: download
Note: This software does not work on Mac or Linux computers.
The Hamamatsu S60 Nanozoomer Slide Scanner is an automated slide scanner. It provides brightfield/histology and fluorescence scanning of whole slides. Training is provided for this scanner.
Objectives: 20X NA=0.8 AIR
Fluorescence Filters:
- 405
- 488
- 555
- mCherry
- CY5
- CY7
Image detection:
- Brightfield/histology: High-speed color CMOS camera
- Fluorescence: Low-noise high-speed CCD camera
Additional Features:
- Z-stacking
- Brightfield/color imaging for histology
Special Rules for Slides:
- Slides must be very clean
- No overhanging coverglass
- No overhanging stickers
- Coverglass must be attached (must not move easily):
- Hard-set mounting media preferred
- Semi-wet/removable gelatin mount
- No wet mount
Instructions to Download Viewer Software:
- Click Here: https://www.hamamatsu.com/jp/en/product/life-science-and-medical-systems/digital-slide-scanner/U12388-01.html
- Scroll down to “Software and Manual Download”
- Download the software for your computer.
Epifluorescent and Brightfield Microscopes
Microscopes with digital cameras for standard epifluorescent/widefield or brightfield samples.
Objectives:
- 10X NA=0.32 Air
- 20X NA=0.55 Air
- 40X NA=0.95 Air
- 60X NA=1.4 Oil
- 100X NA=1.4 Oil
Fluorescence Filters:
- DAPI
- CFP
- YFP
- GFP
- TRITC
- Texas Red
- Cy5
- Cy7
Microscope: Inverted DMi8
Image Detection:
- Brightfield/histology: Leica K3c DOC CMOS camera
- Fluorescence: Leica K8DOC CMOS camera
Additional Features:
- Z-stacking
- Time-lapse with heated chamber
- Mosaic stitching and multi-point visiting
- Automated multi-well imaging
- Phase Contrast
- Polarized Light
- Brightfield/color imaging for histology
- Leica Thunder Deconvolution
Objectives:
- 5X NA=0.16 Air
- 10X NA=0.3 Air
- 20X NA=0.5 Air
- 40X NA=1.3 Oil
- 63X NA=1.4 Oil
- 100X NA=1.4 Oil
Fluorescence Filters:
Standard filter set:
- DAPI
- Alexa 488
- DsRed
- Cy5
Live filter set (upon request):
- CFP
- YFP
- GFP
- TexasRed
- Tripple band pass
Microscope: Inverted Observer Z1
Image Detection:
- Brightfield/histology: Zeiss Axiocam HRc camera
- Fluorescence: Zeiss AxioCam MRm camera
Additional Features:
- Z-stacking
- Mosaic stitching
- Brightfield/color imaging for histology
- Structural Illumination (Apotome)
Lenses: 1x
Magnification Booster: 0.7X to11.5X
Fluorescence Filters:
- DAPI
- GFP
- RFP
- Cy5
Microscope: Upright Olympus SZX16
Image Detection:
- Brightfield/histology: Olympus DP28 color camera
- Fluorescence: Olympus XM10 camera
Additional Features:
- Brightfield/color imaging
- Time-lapse
