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NCI Optical Microscopy Laboratory

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Zeiss LSM 880 Airyscan

Type

Laser scanning confocal microscope

Capabilities

  • Super-resolution confocal imaging – resolution of 140nm, xy and 350nm, z in fixed and live 2D/3D cells
  • Improved sensitivity imaging (Improved signal to noise) – 4x to 8x more sensitive than Zeiss LSM 780 in fixed and live 2D/3D cells
  • Faster imaging – 27 frames per second at 480 x 480 pixels in fixed and live cells
  • Fluorescent Correlation Spectroscopy (FCS)

Laser Lines

  • 405, 458, 488, 514, 561, 594, 633 nm

Objectives

  • 10x, dry, 0.45NA, 2.0 mm
  • 20x, dry, 0.80NA, 0.55 mm
  • 40x, water, 1.2NA, 0.28 mm
  • 40x, oil, 1.3NA, 0.21 mm
  • 63x, oil, 1.4NA, 0.19 mm
  • 63x, multi-immersion, 1.2NA, 0.49 mm
  • 20x, dry, 0.80NA, 0.55 mm
  • 40x, water, 1.2NA, 0.28 mm
  • 40x, oil, 1.3NA, 0.21 mm
  • 63x, oil, 1.4NA, 0.19 mm
  • 63x, multi-immersion, 1.2NA, 0.49 mm

Detectors

  • (2) Multialkali photomultiplier tube (MA-PMT)
  • Spectral Detector – 32 Channel Gallium Arsenide Phosphide Photomultiplier Tube (GaAsP PMT)
  • Airyscan detector – array of 32 individual GaAsP-PMT elements that act as a pinhole size of 0.2 Airy units (AU)

Leica SP8 with Digital Lightsheet

Type

Laser scanning confocal with digital lightsheet microscope

Capabilities

  • 2D / 3D imaging of fixed or live fluorescence-labeled samples
  • Imaging of interactions and dynamics of fluorescence labeled molecules
  • Digital light sheet imaging of cleared large 3D samples both fixed and live
  • Spectral Imaging
  • Forster Resonance Energy Transfer (FRET)
  • Fluorescence Recovery After Photobleaching (FRAP)
  • Laser Lines

Laser Lines

  • 405, 488, 552, 638 nm

Objectives

  • 1.6x, dry, 0.05NA*
  • 2.5x, dry, 0.07NA, 9.2 mm*
  • 5x, dry, 0.15NA, 12 mm
  • 5x, multi-immersion, 0.15NA#
  • 10x, dry, 0.4NA, 2.2 mm*
  • 10x, water, 0.3NA, 3.6 mm#
  • 20x, multi-immersion, 0.75NA, 0.67 mm
  • 25x, water, 0.95NA, 2.5 mm#
  • 40x, dry, 0.85NA, 0.21 mm
  • 63x, water, 1.2NA, 0.30 mm,
  • 63x, oil, 1.4NA, 0.14 mm
  • 100x, oil, 1.4NA, 0.13 mm
  • *Illumination objectives for Digital Lightsheet
  • #Detection objectives for Digital Lightsheet
  • TwinFlect Mirrors for Digital Lightsheet
  • 7.8 mm width; Sample Immersion Liquid – Glycerol, Water; Illumination Obj. – 1.6x, dry; Detection Obj. 5x, multi-immersion or 10x, water
  • 5 mm width; Sample Immersion Liquid -Water; Illumination Obj. – 2.5x, dry; Detection Obj. 10x, water or 25x, water
  • 2.5 mm width; Sample Immersion Liquid -Water; Illumination Obj. – 2.5x, dry or 10x, dry ; Detection Obj. 10x, water or 25x, water

Detectors

  • (1) Photomultiplier tube
  • (2) Hybrid detectors (HyD)

Instructions

Andor spinning disk confocol on Leica DMi8 base

Type

Spinning disk confocal

Capabilities

  • 2D / 3D imaging of fixed or live fluorescence-labeled samples
  • Imaging of interactions and dynamics of fluorescence labeled molecules
  • Large area montages

Laser Lines

  • 405, 445, 488, 514, 561, 594, 640, 730 nm

Objectives

  • 1.6x, dry, 0.05NA
  • 5x, dry, 0.15NA, 12 mm
  • 10x, dry, 0.4NA, 2.2 mm
  • 20x, dry, 0.75NA, 0.62 mm
  • 63x, water, 1.2NA, 0.30 mm,
  • 63x, oil, 1.4NA, 0.14 mm

Detectors

  • Andor Xyla sCMOS camera

Zeiss LSM710

Type

Laser scanning confocal microscope

Capabilities

  • 2D / 3D imaging of fixed or live fluorescence-labeled samples
  • Imaging of interactions and dynamics of fluorescence labeled molecules

Laser Lines

  • 405, 458, 488, 514, 561, 594, 633, 690 – 1000 nm (pulsed)

Objectives

  • 5X, dry, 0.15 NA, 13.6 mm
  • 10X, dry, 0.45 NA, 2.00 mm
  • 10X, water, 0.45 NA, 1.8 mm
  • 20X, dry, 0.4 NA, 7.9 mm
  • 40X, water, 1.1 NA, 0.62 mm
  • 63X, oil, 1.4 NA, 0.19 mm

Detectors

  • Photomultiplier tubes
  • Spectral detector

Instructions

Zeiss LSM780

Type

Laser scanning confocal microscope

Capabilities

  • 2D / 3D imaging of fixed or live fluorescence-labeled samples
  • Imaging of interactions and dynamics of fluorescence labeled molecules
  • Live animal imaging

Laser Lines

  • 405, 458, 488, 514, 561, 633, 690 – 1000 nm (pulsed)

Objectives

  • 10X, dry, 0.3 NA, 5.1 mm
  • 20X, dry, 0.8 NA, 0.55 mm
  • 20X multi-immersion
  • 20X water (dipping lens, not on turret), 1.0NA, 0.19 mm
  • 40X, water, 1.1 NA, 0.28 mm
  • 40X, oil, 1.3 NA, 0.2 mm
  • 63X, oil, 1.4 NA, 0.19 mm

Detectors

  • Photomultiplier tubes
  • Spectral detector
  • Non-descanned detectors (for use with 2P laser)
  • Fluorescence life-time detectors

Instructions

Nikon STORM

Type

Widefield super-resolution microscope for fixed cells only

Capabilities

  • Super-resolution by Point Localization Microscopy – resolution 20nm in xy and 50nm in z.
  • Recommended modality is dSTORM with CF-647 dye (improved version of Alexa Fluor 647 dye).

Laser Lines

  • 405, 488, 561, 647 nm

Objectives

  • 100x oil, 1.49 NA, 0.13 mm WD
  • 100x silicon oil, 1.35 NA, 0.3 mm WD

Cameras

  • Andor iXON DU-897U

Nikon SIM

Type

Widefield super-resolution microscope for fixed cells only

Capabilities

  • N-SIM: Super-resolution Structural Illumination Microscopy (SIM) – resolution of 110nm in xy and 275nm in z.  Compatible with conventional immunofluorescent staining.
  • “External-phase” trans-illumination imaging (on side eye-port camera), which is useful for correlative light/electron microscopy.  None of the high NA objectives on this microscope are compatible with regular phase imaging.

Laser Lines

  • 405, 488, 561, 640 nm

Objectives

  • 10x, dry, 0.3 NA, 16 mm WD
  • 60x water, 1.27 NA, 0.17 mm WD
  • 100x oil, 1.49 NA, 0.13 mm WD

Cameras

  • Andor iXON DU-897E for SIM
  • Nikon DS-QiMC for external phase microscopy

Nikon Multifunctional Microscope (MFM)

Type

Widefield

Capabilities

  • 2D / 3D imaging of fixed or live fluorescence-labeled samples
  • High content analysis
  • TIRF
  • Polarization imaging
  • Simultaneous 2-color imaging
  • Single molecule imaging

Excitation lines

  • (LED illuminator)
  • 395, 440, 470, 508, 555, 640 nm

Objectives

  • 4X, dry, 0.13 NA, 16.5 mm
  • 20x, dry, 0.75 NA, 1.00 mm
  • 60X, oil, 1.40 NA, 0.13 mm
  • 100X, oil, 1.45 NA, 0.13 mm

Detectors

  • Andor NeoZyla sCMOS camera
  • Andor iXon Ultra EMCCD camera

Asylum Research Cypher-VRS

Type

Atomic force microscope

Capabilities

  • Video-rate scanning for molecular dynamics imaging
  • Contact and tapping mode imaging in air and liquid environments
  • Force spectroscopy
  • Mechanical mapping
  • Temperature control
  • Perfusion chamber
  • Semi-automated probe alignment and calibration
  • Built-in analysis tools

Objectives

  • 20X reflective objective for cantilever setup and sample locating

Sample formats

  • Standard imaging: 10 mm diameter substrate
  • Video-rate imaging: 3 mm diameter substrate

Celigo

Type

Plate reading microscope

Capabilities

  • Built in analysis tools
  • Fast, widefield imaging of entire well for multiwell plate
  • Built in protocols for live/dead counting, growth curves, spheroid analysis etc.

Excitation/Emission filters

  • Blue 377/477
  • Green 483/536
  • Red 531/629
  • Far Red 632/692
  • Brightfield

Objectives

  • 1 objective optimized for entire well imaging & built in analysis features

Sample formats

  • Works best with supported/recommended plates
  • Microwell plates: 6, 12, 24, 48, 96, 384, 1536 wells
  • Slides
  • 10 cm Petri Dish
  • T25 Flasks

Instructions