Confocal Microscopy

Andor Dragonfly - Confocal / TIRF / Super Resolution

The Dragonfly microscope is a fully automated multimodal microscope that can be used for confocal, total internal reflection microscopy (TIRF) and super-resolution (localization-based techniques such as: PALM, dSTORM, PAINT, etc.).

 

Applications:

  • Spinning disk confocal imaging. In contrast to point scanning confocals, such as the LSM880 and CD7 that serially acquire one focal point at a time to reconstitute a 2D image, the Dragonfly acquires confocal images by simultaneously acquiring thousands of spots from the focal plane. This setup enables the Dragonfly to provide several important advantages over point-scanning confocals:
    • Faster speed (up to ~200 frames per second), which is ideal for fast 2D or 3D time-series or tile scanning.
    • Gentler imaging – photobleaching and phototoxicity are greatly reduced, which is better for live cell imaging.
    • The high sensitivity of cameras is well suited for dim specimens.
  • Super-resolution imaging. The Dragonfly can perform 3D localization- based microscopy techniques (dSTORM, PAINT, etc.) that improve spatial resolution to ~30nm laterally, and ~50nm axially.
  • TIRF microscopy. a technique allowing the confined excitation of fluorophores within ~100nm above the coverslip.

Objectives: 10x/0.45, 20x/0.8, 40x/1.1 W, 40x/1.3 Oil, 63x/1.47 Oil.

Laser lines: 405nm, 488nm, 561nm, 638nm, 735nm.

Detectors: Two Andor Zyla 4.2 sCMOS cameras can be used together to acquire two colors simultaneously.

Operating software: Fusion

Location: APB 141R

Contact persons: Dr. Le Marchand (confocal), Tim Chaya (super-resolution)

Zeiss Celldiscoverer7

The Celldiscoverer 7 is an automated boxed microscope with a user-friendly interface that allows wide-field, confocal (LSM 900), and super-resolution imaging by means of its Airyscan2 detector.  The system is an inverted microscope capable of live cell imaging experiments with a variety of incubation conditions.  It has enhanced automated functionality such as Autocorr objectives, which enable the system to automatically detect a variety of sample holders and adjust for spherical aberrations. The CD7 is also equipped with an autoimmersion system that automatically applies water to its water immersion lenses and maintains the volume of immersion water to permit live cell imaging over a long duration.  The system uses LED illumination, which has low phototoxicity and long-term stability.

Applications:

  • Live cell imaging
  • High-throughput imaging
  • Tiling
  • Time-lapse
  • Optical sectioning for 3D imaging
  • Super resolution imaging with the Airyscan 2
  • FRAP and FRET

Objectives:

  • PlanApochromat 5x/0.12
  • PlanApochromat 20x/0.7 and 20X/0.95, Autocorr
  • PlanApochromat 50x/1.2 water, Autocorr and autoimmersion

These objectives are used in combination with a 0.5X, 1x and 2x optovar lens, which permits 2.5x, 5x, 10x, 20x, 25x, 40x, 50x, 100x magnifications.

Laser Lines:

  • 405 nm, 488 nm, 561 nm, and 640 nm

Detectors:

  • For LSM900 Confocal:
    • Airyscan 2 detector
    • 2 PMT detectors
  • For Wide-field Imaging:
    • Axiocam 506 black and white camera (2752 × 2208 pixels)

Stage Inserts:

  • The system accommodates most multiwell plates, 3.5 and 6 cm petri dishes, microscopy slides, and chamber slides

Operating Software: Zen Blue

Location: APB 141T

Contact Persons: Dr. Chandran Sabanayagam and Dr. Jeffrey Caplan

Zeiss 880 at APB - Confocal / Spectral / Multiphoton / Airyscan

The LSM 880 microscope is a fully automated multimodal microscope that can be used for confocal, spectral, and multiphoton excitation imaging. The Airyscan detector enables super-resolution and high speed imaging.

The LSM880 is fully equipped for live-cell imaging. It has an environmental enclosure (temperature, humidity, and CO2 control) and z-drift correction to maintain samples at the focal plane over time.

 

Applications:

  • Confocal imaging.
  • Spectral imaging with linear unmixing. It can be used to remove background autofluorescence or for multicolor imaging when spectral overlaps between colors cannot be separated with standard confocal microscopy.
  • Multiphoton excitation microscopy. It can be used to image up to 10x deeper than confocal microscopy.
  • Super-resolution imaging. The Airyscan can provide a 2-fold increase in resolution compared to conventional confocal microscopy.
  • High-speed imaging. The Airyscan detector can be used to image 4 times faster than the confocal mode (up to 20fps at 512*512). The 880 is also equipped with a piezo-electric stage for high-speed acquisition of z-stacks.
  • Confocal reflection microscopy. It can be used to image collagen fibers and surface microtopography.
  • FRET, FRAP, FLIP.
  • Fluorescence Correlation Spectroscopy imaging. It provides quantitative data on concentrations, diffusion coefficients, molecular transport and interactions.
  • Tile scanning to image a large field at high resolution.
  • Live cell imaging thanks to the environmental enclosure.

Objectives: 10x/0.3, 20x/0.8, 40x/1.2 W, 40x/1.3 Oil, 63x/1.4 Oil – more objectives are available as add-ons.

Laser lines: 405nm, 458nm, 488nm, 514nm, 561nm, 633nm, 680-1080nm.

Detectors: a 32-channel ultra-sensitive spectral GaAsP PMT (Gallium Arsenide Phosphide PhotoMultiplier Tube) detector (i.e. ChS in Zen) which is ideal for spectral imaging, two multi-alkali PMT detectors (Ch1 & Ch2), one transmitted PMT (T-PMT) for contrast imaging (Bright Field, Differential Interference Contrast, Phase), and one GaAsP Airyscan detector for super-resolution and fast imaging.

Operating software: Zen 2.3 SP1

Location: APB 141S

Contact person: Dr. Le Marchand

 

Zeiss 880 at Wolf Hall - Confocal / Spectral / Airyscan

The LSM 880 microscope is a fully automated multimodal microscope that can be used for confocal and spectral imaging. The Airyscan detector enables super-resolution and high speed imaging.

The LSM880 is fully equipped for live-cell imaging. It has an environmental enclosure (temperature, humidity, and CO2 control) and z-drift correction to maintain samples at the focal plane over time.

Applications:

  • Confocal imaging.
  • Spectral imaging with linear unmixing. It can be used to remove background autofluorescence or for multicolor imaging when spectral overlaps between colors cannot be separated with standard confocal microscopy.
  • Super-resolution imaging. The Airyscan can provide a 2-fold increase in resolution compared to conventional confocal microscopy.
  • High-speed imaging. The Airyscan detector can be used to image 4 times faster than the confocal mode (up to 20fps at 512*512). The 880 is also equipped with a piezo-electric stage for high-speed acquisition of z-stacks.
  • Confocal reflection microscopy. It can be used to image collagen fibers and surface microtopography.
  • FRET, FRAP, FLIP.
  • Fluorescence Correlation Spectroscopy imaging. It provides quantitative data on concentrations, diffusion coefficients, molecular transport and interactions.
  • Tile scanning to image a large field at high resolution.
  • Live cell imaging thanks to the environmental enclosure.

Objectives: 5x/0.25, 10x/0.3, 20x/0.8, 40x/1.2 W, 40x/1.3 Oil, 63x/1.4 Oil.

Laser lines: 405nm, 458nm, 488nm, 514nm, 561nm, 594nm, 633nm.

Detectors: a 32-channel ultra-sensitive spectral GaAsP PMT (Gallium Arsenide Phosphide PhotoMultiplier Tube) detector (i.e. ChS in Zen) which is ideal for spectral imaging, two multi-alkali PMT detectors (Ch1 & Ch2), one transmitted PMT (T-PMT) for contrast imaging (Bright Field, Differential Interference Contrast, Phase), and one GaAsP Airyscan detector for super-resolution and fast imaging.

Operating software: Zen 2.3 SP1

Location: Wolf Hall #304

Contact person: Dr. Le Marchand

Zeiss 780 Multiphoton Confocal

The Zeiss LSM 780 is an upright confocal (objectives are above the sample) that enables confocal, spectral, and multiphoton excitation imaging. The non-descanned detection system can be used for two-photon excitation fluorescence or detection of second harmonic generation (SHG) in the forward and reverse directions. The 780 is particularly well-suited for intravital imaging of small animals.

Applications:

  • Confocal imaging.
  • Spectral imaging with linear unmixing. It can be used to remove background autofluorescence or for multicolor imaging when spectral overlaps between colors cannot be separated with standard confocal microscopy.
  • Multiphoton excitation microscopy. It can be used to image up to 10x deeper than confocal microscopy. The upright configuration, non-descanned detection system and the Z-deck add-on are ideal for intravital imaging of small animals. The system is also ideal for reverse and forward detection of second harmonic generation (e.g. can be used to image collagen fibers).
  • Confocal reflection microscopy. It can be used to image collagen fibers and surface microtopography.
  • FRET, FRAP, FLIP.
  • Fluorescence Correlation Spectroscopy imaging. It provides quantitative data on concentrations, diffusion coefficients, molecular transport and interactions.
  • Tile scanning to image a large field at high resolution.

Objectives: 10x/0.3, 20x/0.75, 40x/1.2 W, 40x/1.4 Oil. Other objectives available in the room: 20x/0.6, 20x/1 W, 40x/0.8 W, 63x/0.9 W, 63x/1.4 Oil.

Laser lines: 405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 633 nm, 690-1040nm.

Detectors: a 32-channel ultra-sensitive spectral GaAsP PMT (Gallium Arsenide Phosphide PhotoMultiplier Tube) detector (i.e. ChS in Zen), two multi-alkali PMT detectors (Ch1 & Ch2), two non-descanned GaAsP PMT detectors, and one transmitted PMT (T-PMT) for contrast imaging (Bright Field, Differential Interference Contrast, Phase).

Operating software: Zen 2.3 SP1

Location: CBBI (Center for Biomedical and Brain Imaging) #244C

Contact person: Dr. Le Marchand

Zeiss 5 LIVE DUO Highspeed Confocal / Spectral

The Zeiss 5 LIVE DUO is line-scanning microscope and has all of the capabilities of a standard confocal. The high speed DUO detector is optimal for capturing fast cellular dynamics and can collect images at 120 frames per second. The META is a spectral detector that can separate fluorescent compounds with similar spectral profiles.

A stage top incubator is available for live cell imaging.

Laser Lines: 405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 633 nm

Location: DBI 117Z

Contact person: Dr. Caplan