Light Microscopy

 Fluorescent/Brightfield Microscopy

Leica DM6B with Deconvolution Software

The Leica DM6 B upright microscope is equipped with Hamamatsu sCMOS camera, Leica DFC450 color CCD, Spectra X LED light engine, and LAS X extended software package.

  • Differential Interference
  • Contrast
    Polarizing Imaging

  • Metal halide lamp and filter sets
    • DAPI/UV: Ex 325-375/Em 435-485
    • GFP: Ex 450-490/Em 500-550
    • TRITC: Ex 532-558/Em 570-640
    • TXR: Ex 540-580/Em 592-668
    • Cy5: Ex 590-650/Em 662-738
    • Cy7: Ex 672-747/Em 765-855
  • Quad Sedat EFW
    • DAPI/UV: Ex 325-375/Em 430-480
    • FITC: Ex 480-500/Em 507-543
    • TRITC: Ex 542-568/Em 579-631
    • Cy5: Ex 630-660/Em 669-741
  • Fura
    • Fura2-ET EFW: Ex 340 and 380/Em 470-550

  • Dry: 5x/0.15, 10x/0.30, 20x/0.80, 40x/0.85
  • Water immersion: 20x/0.75, 40x/0.80
  • Oil immersion: 40x/1.25-0.75, 40x/1.30, 63x/1.40-0.60, 63x/1.40, 100x/1.4-0.70, 100x/1.30

Equipped with QE enhanced Orca Flash 4.0 V2 sCMOS camera and LAS X extended software for digital image capture and analysis

  • Multichannel acquisition and image overlay with labeling
  • Constrained iterative deconvolution in 2-D and 3-D
  • 3-D reconstruction (surface and volume)
  • Time lapse
  • Montage image
  • Analysis, graphs and spreadsheets of data
  • Resolution of camera 2048 x 2048 pixel
  • 100 frames per second (fps) at full resolution in Standard Scan mode or 30 fps in Slow Scan mode

Files can be opened using the software LAS X Office and Fiji ImageJ.

Leica DM6BLeica DM6B in Chemistry & Materials Building, Room 155

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Leica Thunder Model Organism Stereomicroscope

Leica MC205FA stereo/macroscope. Fully automated Z drive with 420 mm range and 1 µm z-resolution. Manual zoom from 0.78 to 16×. Four position filter turret; 2 coded filter sets. High precision scanning stage with integrated controller; position resolution ≤ 5 nm and maximum load capacity of 1.5 kg.

  • Fluorescent multi-channel imaging
  • Brightfield
  • Darkfield
  • Live imaging
  • Tiled imaging
  • Point scanning
  • Multiwell and petri scanning
  • Timelapse
  • Deep imaging
  • Fast acquisition
  • On the fly deconvolution
  • Adaptive deconvolution.

  • ET GFP Ex 450-490/Em 500-550
  • ET mCHER – Ex 540-580/Em 593-667

  • Leica FLEXACAM C1 with 12 MP CMOS sensor
  • Leica K5 with 4.2 MP sCMOSs sensor

  • PLANAPO 1.0.x, working distance 61.5mm
  • PLANAPO 5.0x/0.50 LWD working distance 19mm

  • Leica TL5000 Ergo Transmitted Light Base
  • Leica LED5000 RL-80/40
  • Leica LED3 (fluorescence), wavelength white 390-680 nm

  • Multichannel fluorescence acquisition
  • Z-stack capture
  • Time lapse
  • Measurement and Data Analysis

Files can be opened using the software LAS X Office and Fiji ImageJ.

Leica Thunder

Leica Thunder Model Organism Stereomicroscope in Chemistry & Materials Building, Room 155

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 Fluorescence Confocal Microscopy

Leica TCS SP8 Confocal Microscope with WLL

Leica SP8 Confocal White Light Laser system operates from a DMI-8 CS Bino inverted microscope with AFC closed-loop focusing, scanning, and Super Z-Galvo stages, turret cooling, spill protection, and CUDA workstation motorized DIC accessories.

  • UV / 405 nm
  • Argon – Blue (458, 476, 488, 496, and 514nm)
  • White Light Laser system (470 to 670 nm + Ar) tunable in steps of 1 nm

5 channels of Confocal Detection

  • 3 Super Sensitive Spectral Hybrid detectors
  • 2 cooled PMT detectors

  • Tandem Scanner (Variable and Resonant) for imaging at speeds from 1 to 16000Hz
  • Scanning XY-stage for montage imaging
  • Automated DIC for all installed objectives (10x/0.30 dry; 20x/0.75 and 63x/1.20 water immersion; 40x/1.30 and 63x/1.40 oil immersion)
  • Adaptive Focus Control
  • LightGate Imaging via pulsed WLL and HyD detectors helps remove reflected light as well as autofluorescence
  • HyVolution 2 integrated deconvolution during image acquisition for improved contrast and resolution
  • Z-stack for 3-D rendering
  • Brightfield
  • Transmission
  • Reflection
  • Fluorescence
  • Multi-color simultaneous acquisition
  • Serial acquisition
  • Time-lapse
  • FRET
  • FRAP

Files can be opened using the software LAS X Office and Fiji ImageJ.

Leica SP8Leica TCS SP8 Confocal Microscope with WLL in Chemistry & Materials Building, Room 151.

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Olympus SpinSR10 Spinning Disk Confocal 

Built on an IX83 base, the SpinSR10 is a fantastic spinning disk confocal and excels at rapid image capture. This system utilizes super-resolution components as well as the optical sectioning capability of confocal fluorescence microscopes. The system simultaneously achieves high reliability, fast image acquisition, and low phototoxicity, which cannot be done using conventional methods. Based on optical theory, Olympus Super Resolution (OSR) processing algorithms can obtain a spatial resolution of 120 nm, providing highly reliable results below the diffraction limit. Fast image acquisition is achieved using the spinning disk confocal microscope technique. Using the available super-resolution SoRa disk technology, phototoxicity can be reduced to 30% of conventional techniques. Equipped with an incubation system with temperature and CO2 control for long-term in vivo experiments.

  • Two Hamamatsu ORCA-Fusion (16-bit grayscale)

Description
Power (mW)
Wavelength (nm)
LD405 50 405 
LD488 100 488
LD561 100 561
LD640 100 640

 

  • 50µm
  • SoRa

  • D405/488/561/640
  • D445/514/640

  • Filter wheel 1:  B447/60; B525/50; B617/73; B685/40; B535/30
  • Filter wheel 2:  B447/60; B525/50

Magnification
Objective Type
Description
NA
Refraction Index
WD (mm)

4

 UPLFLN PH 4x 0.13 AIR (1.000) 17.00
10 UPLXAPO 10x 0.40 AIR (1.000) 3.10
20 UPLXAPO 20x 0.80 AIR (1.000) 0.60
40 UPLXAPO O 40x 1.40 OIL (1.518) 0.13
100 UPLAPO OHR 100x 1.50 OIL (1.518) 0.12

 

Files can be opened using Fiji ImageJ.

Olympus SpinSR10 Spinning DiskOlympus SpinSR10 Spinning Disk Confocal in Chemistry & Materials Building, Room 151

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Light Sheet Microscopy

Zeiss Light Sheet 7

Light Sheet Fluorescence Microscopy (LSFM) is an extremely powerful alternative to established fluorescence imaging techniques, especially for 3-D imaging within whole live organisms and large tissue explants. Ideal for fast and gentle imaging of whole living model organisms, tissues, and cells as they develop – over extended periods of time capable of imaging large optically cleared specimens in toto – with subcellular resolution. By selectively illuminating the observed optical section with a thin sheet of light, photo bleaching is reduced to a minimum, making light sheet microscopy ideal for nondestructive imaging of fragile samples over extended periods of time. Equipped with an incubation system with temperature and CO2 control for long-term in vivo experiments.

 Magnification
Objective Type
Description
NA
Immersion
WD (mm)
2.5 Fluar 2.5x 0.12 Clearing 8.7
5 EC Plan-Neofluar 5x 0.16 foc Water/Clearing 18.5
5 EC Plan-Neofluar 5x 0.16 Clearing 18.5
10 W Plan-Apochromat 10x 0.50 Water 3.7
20 W Plan-Apochromat 20x 1.00 Water 2.4
20 W Plan-Apochromat 20x 1.00 Clearing 6.4

 

  • 405
  • 445
  • 488
  • 514
  • 561
  • 638

Fluorochromes
Mirror
Filter 1 (nm)
Filter 2 (nm
DAPI/GFP SBS LP 490  BP 420-470 (DAPI) BP 505-545 (GFP)
DAPI/RFP SBS LP 510 BP 420-470 (DAPI)  BP 575-615 (RFP)
GFP/mCherry SBS LP 560 BP 505-545 (GFP)  LP 585 (mCherry)
GFP/ Draq5(toto3) SBS LP 560 BP 505-545 (GFP)  LP 660 (Draq5(toto3))
RFP/ Draq5(toto3) SBS LP 640 BP 575-615 (RFP)  LP 660 (Draq5(toto3))

 

  • Two liquid cooled sCMOS cameras, pco.EDGE

  • Red      =          0.68 mm
  • Black    =          1.00 mm
  • Green   =          1.50 mm
  • Blue     =          2.15 mm

Zeiss Ligth Sheet 7

Zeiss Light Sheet 7 in Chemistry & Materials Building, Room 155

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