Instruments

N-SIM Super-Resolution, Dual Camera System

Structured illumination based super-resolution microscope delivering twice the resolution of traditional diffraction limited microscopes. N-SIM offers temporal resolutions of up to 0.6sec/frame, providing an ideal platform for live-cell imaging at super-resolution. Various imaging modes are available, including TIRF-SIM, 2D-SIM and 3D-SIM. In addition, Princeton’s N-SIM system is equipped with an emission splitter and two cameras enabling simultaneous two color super-resolution imaging.

Nikon A1 and Nikon A1-RS Confocal Microscopes

  • At the heart of the Nikon A1 hybrid confocal scan head is the dual scanning system that encompass a standard-speed scanner and a high-speed resonant scanner. They may be used individually or simultaneously. Applications include:
  • For the standard speed scanner: imaging of all fixed specimens and many live specimens, including FRET and ion ratio imaging (e.g.: calcium, pH);
  • High speed resonant scanner: Maximum frame rate can exceed 400 frames per second.
  • Simultaneous use of standard and resonant scanners: Useful for FRAP and photoactivation specimens that require stimulating with one scanner while simultaneously imaging with the other.
  • Three Detector Ports are Standard
  1. Standard Detector. The standard filter-based detector allows simultaneous or sequential detection of up to four channels of fluorescence.
  2. Spectral Detector. With the 32-channel spectral detector, up to eight individual overlapping fluorophores can be unmixed simultaneously and acquired at a maximum frame rate of 16 frames per second.
  3. A transmitted light detector allows simultaneous capture of a DIC image while capturing confocal fluorescence images.

Laser Platform:

  • 405nm diode laser, 561nm diode-pumped-solid-state (DPSS) laser, and a 638nm diode laser.
  • An argon-ion gas laser is the fourth laser. It produces four lines: 457nm, 477nm, 488nm, and 514nm line.
  • Nikon Perfect Focus System (PFS) is the solution to keep cells in focus over prolonged time-lapse imaging sessions. Pick the focal plane and PFS monitors the focus every 5ms to maintain focus indefinitely.
  • Accessories: Motorized XY Stage and Piezo Z stage for rapid sectioning.
  • Stage top incubator for temperature, humidity, and CO2 atmospheric control.

Leica SP5

  • The Leica SP5 Confocal has a filter-free spectral detector system with five individually tunable channels. The spectral separation is accomplished by a unique prism design. 
  • Each detector is freely tunable over the entire visible spectrum.  The SP5 has a prism base beam splitter (AOBS) that allows for free selection of up to 8 Lasers from UV – IR.  The spectral range of the detector optics covers 400 - 800nm.
  • The scanner can be run in either conventional or resonant modes. The conventional scanner can be scanned unidirectional or bi-directional with a speeds ranging from 10Hz to 2800Hz.  The resonant scan can also be scanned unidirectional or bi-directional at 8000Hz or 1600Hz respectively with a maximum frame rate of 250.  
  • The system is equipped with a Z-galvanometer that allow for XZ scanning as well as fast XYZ scanning.  The scanning stage allows for stitching together images from large fields of view as well as imaging multiple positions as XYZT images.
  • The laser lines available on the SP5 are 405, 458, 476, 488, 514, 561, 594 and 633.  Of the 5 detectors on the system 3 are conventional PMTs, 2 are Hybrid detectors (HyD).  Leica HyD detectors provide highly sensitive photon detection.
  • There is a Warner chamber for temperature control of live experiments

Nikon Ti-E with Spinning Disc and Ablation Module

  • The microscope base is a fully motorized Nikon Ti-E with Perfect Focus System (interferometric based focus maintenance (PFS)). The system also has a piezo stage with 100 um range capable of up to 100 steps/sec, mounted in a fully motorized XY stage.
  • A Yokogawa spinning disc (CSU-21) is mounted with a quad dichroic accommodating 405, 488, 561, and 647 lasers. The spinning disc detector is a Hamamatsu ImageM back thinned EMCCD (512, 93% QE, 30 full fps) and an Orca Flash fitted with a TuCam adapter.
  • A 100 mW 285 MicroPoint ablation module is mounted, capable of producing 365 nm ablation, 405 nm for PA/PC, and 435 nm for FRAP experiments.
  • This system has an Agilent laser launch with 405, 488, 561, and 640 lasers.
  • There is also an environmental control chamber for long-term live cell imaging.

Nikon TIRF and Super Resolution (localization type)

  • Nikon Ti-E with PFS fitted with motorized TIRF module with mag optic and ¼ wave for super resolution localization applications.
  • An Agilent high power MLC400 provides;
    • 150 mW    647 nm
    • 70 mW      561 nm
    • 70 mW      488 nm
    • 25 mW      405 mW
  • In addition to TIRF capabilities, this system has been optimized for super resolution capabilities (localization type).
  • There is a 512 back-thinned EMCCD (Andor iXon Ultra) fitted on to this system.
  • A stage piezo also allows for very fast z section acquisition.
  • There is also an environmental control chamber for long-term live cell imaging.

Nikon Widefield Platform

  • Nikon TE2000 fitted with Andor Zyla sCMOS camera and a color Nikon Ri2 16.5 MP camera. This system is capable of acquiring basic widefield images, transmitted (DIC) and epifluorescent.
  • There is also a motorized stage to acquire large tiled or multipoint images.

Perkin Elmer Operetta

  • The Operetta is a high content plate imaging system.
  • Fast widefield imaging for routine applications and confocal imaging option to eliminate background, improve signal-to-noise and protect live cell samples by minimizing photobleaching and phototoxicity
  • Unsupervised automated image acquisition and autofocus for reliable high content imaging from slides to multiple plates.

Prairie ultima multi-photon system

  • 2 channel imaging using 2 GaAsp detectors per channel. A Coherent Chameleon Ultima II pulse laser, tunable from 680 to 1080 nms.
  • Dual scanning system - galvo scanners and an AOD scanner with scan speeds up to 25 fps at 512 x 512 resolution
  • Fast Prior piezo z-stage with 500 microns of travel
  • 405 laser for ablation, photo-activation and photo-conversion
  • Environmental chamber for live cell mammalian imaging. Temp. Range 25 to 45 degrees C.\
  • Objective lenses: 1.0 NA20 x and 40 x Plan Apo's, 1.3 NA 40 x Plan Apo
  • 0.2 NA 5X Plan fluor and 10 x 0.6 NA Plan Apo