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Washington University in St. Louis

Washington University Center for Cellular Imaging

The fundamental goal of the Washington University Center for Cellular Imaging (WUCCI) is to provide access to an integrated infrastructure of light and charged particle based cellular imaging technologies. The Center Director, and his staff provide both professional guidance and work collaboratively with Washington University researchers in assay design, sample preparation and data analysis as well as develop and apply new imaging approaches and informatics methods. The Nikon Center of Excellence within the WUCCI enables investigators to gain unprecedented insights into the dynamic behavior of single molecules and the spatial organization of cells and tissues using state-of-the-art confocal, live-cell and super-resolution microscopies, creating exciting opportunities for discovery in a broad range of basic and translational research aimed at advancing our understanding of human health and disease.

 

Systems Available

A1Rsi with a Ti-E Inverted Microscope

The Nikon A1Rsi is a scanning confocal microscope is suitable for a broad range of applications including spectral and rapid imaging. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the Perfect Focus stability mechanism. The system is equipped with a unique hybrid scan head, incorporating both a high-resolution galvo scanner and an ultra-high speed resonant scanner, enabling simultaneous photo-bleaching / photo-activation and image acquisition. The scan head is also equipped with two GaAsP PMTs, and two high-sensitivity PMT detectors for four-color confocal imaging as well as a transmitted light PMT for simultaneous DIC imaging. The enhanced spectral acquisition capabilities include a 32-channel detector for spectral acquisition at full image (512 x 512 pixel) resolution in 0.6 seconds.

System Configuration

  • Laser lines: 405, 445, 488, 514, 561 and 640nm
  • Objectives: 10X and 20X dry and 40X, 60X and 100X high NA oil immersion
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity, CO2 and O2 regulation

Applications

  • Multi-color confocal microscopy
  • DIC transmitted light microscopy
  • Three-dimensional reconstructions
  • Spectral imaging / separation
  • Co-localization analysis
  • FRAP / FRET analysis
  • Photo-activation / photo-stimulation
  • Multi-position imaging / tiling
  • Live Cell imaging (T, H, CO2 and O2)

 

 


Nikon Spinning Disk Confocal Microscope

The Nikon Spinning Disk (SD) confocal microscope is designed for high-magnification 4D imaging of living cells with minimal effects of photo-damage. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the Perfect Focus stability mechanism. The system is fitted with a Yokagawa CSU-X1 variable speed Nipkow spinning disk scan head for the confocal illumination and imaging of an entire field of view (FOV) simultaneously on a camera. The scan head is equipped with an Andor TuCam two-camera adapter that enables simultaneous two-color confocal imaging at video-rate (30fps) on two separate Andor Zyla 4.2 Megapixel sCMOS cameras. The system also boasts an LED-based DMD system for ultrafast photo-stimulation and a separate galvo-based laser scanner for slower FRAP and photo-activation studies.

System Configuration

  • Laser lines: 405, 488, 561 and 640nm
  • Objectives: 10X and 20X dry, 60X and 100X high NA oil immersion and 40X water immersion (corr).
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Ti LAPP DMD (Deformable Mirror Device) LED source for ultrafast photo-stimulation
  • Miniscanner galvo-based laser scanning system for FRAP / photo-activation
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity, CO2 and O2 regulation

Applications

  • Multi-color high-speed confocal microscopy
  • DIC transmitted light microscopy
  • Three-dimensional reconstructions
  • FRAP analysis
  • Photo-activation / photo-stimulation
  • Multi-position imaging / tiling
  • Live Cell imaging (T, H, CO2 and O2)

 

 


N-SIM Super-Resolution Structured Illumination Microscope

The Nikon N-SIM microscope allows for sub-diffraction super-resolution imaging using high-frequency structured illumination in three-dimensions for resolutions beyond 100nm. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the Perfect Focus stability mechanism. Fluorescence detection is provided by two Andor Zyla 4.2 Megapixel sCMOS cameras fitted to an Andor TuCam dual camera adapter, which facilitates simultaneous two-color and sequential four-color super-resolution imaging.

System Configuration

  • Laser lines: 405, 488, 561 and 640nm
  • Objectives: 60X water immersion and 100X oil immersion high NA lenses
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity and CO2 regulation

Applications

  • Multi-color super-resolution microscopy
  • Three-dimensional super-resolution reconstructions
  • Super-resolution co-localization
  • Multi-position imaging / tiling

 

 


N-STORM Super-Resolution TIRF Microscope

The Nikon N-STORM TIRF Microscope allows imaging close to the surface of a coverslip facilitating imaging of endo- and exo-cytic processes as well membrane trafficking events. The system is based on a motorized inverted Nikon Ti-E microscope frame equipped with the Perfect Focus stability mechanism. Fluorescence detection is provided by either an Andor Zyla 4.2 Megapixel sCMOS camera when imaging in TIRF mode or an Andor iXon 897 EM-CCD when imaging in single-molecule or STORM (STochastic Optical Reconstruction Microscopy) super-resolution imaging mode.

System Configuration

  • Laser lines: 405, 488, 561 and 640nm
  • Objectives: 10X and 20X dry and 60X and 100X oil immersion high NA lenses
  • Motorized stage: xy-stage including a nano-positioning piezo z-insert
  • Incubation: Tokai-hit stage-top incubator capable of temperature, humidity and CO2 regulation

Applications

  • Multi-color STORM super-resolution microscopy
  • 3D STORM super-resolution microscopy
  • Single-molecule imaging
  • FRET / smFRET
  • Super-resolution co-localization
  • Multi-position imaging / tiling

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