Nikon A1R MP

UF | ICBR Cytometry

Nikon A1R MP Confocal/Multiphoton/STORM Microscope

Laser-scanning confocal (LSCM), multiphoton (MP), and STORM (super-resolution) inverted microscope for fluorescent samples. Available Tokai-Hit stage top incubator with active CO2 and temperature regulation, and passive humidity, for imaging live cells in a 35mm dish or a chambered coverglass. Motorized XY stage movement for multi-point time-lapse imaging or tile-mapping (AKA mosaic or stitching). Galvo and resonant scanning. Software: Nikon NIS-Elements v5.0 on a Windows 7 workstation. (This software is also available (with the optional Deconvolution module) on our separate Windows 7 Analysis Workstation.)

Confocal Mode:  Six visible laser lines: 405, 445, 488, 514, 561 and 647nm. Objectives: 10x dry, 20x dry, 40x dry, 60x water, 60x oil and 100x oil. Primary fluorescent detector unit: filter-based four channel detector configured with DAPI, CFP, GFP, YFP, TRITC/TxRed and CY5 filters. Optional 32-channel spectral detector unit.
Multiphoton (MP) Mode:  Spectra-Physics Mai-Tai IR variable wavelength pulse laser: 690-1040nm. Objectives: 60x water, 60x oil and 2mm working distance 25x water. NOTE: The system must be disassembled and reconfigured to mount the 25x MP objective. Therefore, use of the 25x objective must be coordinated with CTAC staff in advance. Three NDD channels configured for either DAPI-GFP-RFP, or DAPI-CFP-YFP combinations.
STORM Mode:  Camera based super-resolution down to 20nm. Requires the use of specific fluorophores and buffer media. Utilizes TIRF style evanescent wave for fluorescent excitation for a maximum imaging Z-depth of <2um from the edge of the coverslip. Hamamatsu ORCA-Flash 4.0 sCMOS monochrome camera. Special 100x HP STORM objective. High-power 125mw 647nm laser.

PLEASE NOTE: The NIH requires that any papers or abstracts that utilize images or data generated on this instrument must cite NIH Grant # 1S10OD020026 in the acknowledgements. Send a copy of the final published version of any papers, review articles or abstracts that used images or data generated on this instrument to Dr. Habibeh Khoshbouei habibeh@ufl.edu and Doug Smith dugsmith@ufl.edu.

The Nikon A1RMPsi-STORM4.0 imaging solution is a turnkey system with a number of unique features that will enhance the end-user’s research. Such a turnkey multiphoton and super resolution imaging solution allows UF researchers to 1) image deeper than traditional confocal limitation, 2) reconstruct super resolution structure, and also 3) utilize the system for single-photon confocal imaging.

Some of the capabilities on this impressive and versatile system listed below:

  • High Resolution single-photon excitation confocal imaging
  • Fast confocal imaging with a resonant scanners
  • Confocal hybrid (galvano/resonant scanners) mode for advanced applications such as photostimulation
  • and photoactivation
  • Multiphoton imaging with either or both scanners with a tunable multiphoton laser (700nm-1080nm)
  • TIRF imaging with a Hamamatsu Flash 4.0 sCMOS camera for high resolution and large FOV
  • The new N-STORM 4.0 for super resolution imaging, suitable for both live-cell and fixed imaging, is
  • capable to resolve 20nm in lateral resolution and 50nm in axial resolution

A1RMPsi-STORM4.0 General system description:

  • Visible wavelengths for single-photon and TIRF imaging:
    • LU-NV single laser launch with diode lasers: 405nm(20mW), 445nm(20mW), 488nm(70mW), 515nm(40mW), 561nm(70mW), and 647nm(125mW)
    • 4 channel simultaneously: 405nm/488nm/561/640nm
    • All combinations of wavelengths are possible in sequential mode!
    • Transmitted light detector for a widefield overlay in addition to the fluorescence confocal
  • Multiphoton imaging with a Spectra-Physics Mai Thai DS laser
    • This mode can integrate the 405nm laser for photoconversion!
    • Spectral detector to unmix overlapping fluors
  •  Widefield imaging with the all-LED Lumencor Sola Light Engine (365nm-650nm)
    • Hamamatsu Flash 4.0 sCMOS camera (up to 500 FPS)
    • Filters available:
      • DAPI, CFP, GFP, YFP, DS Red, TxRed, Dual GFP/TxRed, CY5
  • TIRF imaging with LU-NV laser launch
  •  N-STORM 4.0 Super Resolution
    • Hamamatsu Flash 4.0 sCMOS camera (up to 500 FPS)
    • Imaging channels (488/561/647nm
    • Resolution – 20nm lateral, 50nm axial

Manual:

Location: CTAC Imaging Suite (MBI LG-164)

 

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