Imaging and Cell Analysis Core

Core Instruments

Four confocal microscopes are available: Leica SPE, Nikon A1R, standard Leica SP8, and Leica Falcon SP8 with a high-speed resonant scanner, fluorescence lifetime imaging (FLIM) capability and deconvolution software.

Training for Instrument Use

All new imaging core users must be trained on confocal, TIRF or Keyence microscopes before signing up to use them. For training on the Nikon confocal microscope, please contact Alice Meng. To use the Nikon TIRF microscope, please contact Dr. Selene Lomoio. For use of any Leica instrument, please contact Dr. Martin Hunter. To be trained on the Keyence microscope, please contact Aislinn Keane. Access imaging core sign-up calendars here.

Contact

  • For access to Stearns 207A, contact Alison Hochler.
  • For issues with Leica instruments or to modify calendar reservations for any instrument, contact Dr. Moritz Armbruster..
  • For help with the Nikon A1R, please contact Alice Meng.

Co-Directors
Rob Jackson, PhD
Michele Jacob, PhD

Location
Stearns 207

Please follow the links below or contact core personnel for information about services and fees.

  • The current fee structure for academic users of the confocal and TIRF microscopes is as follows:

    • Leica SP8  Falcon - $50/hour
    • Leica SP8 - $50/hour
    • Leica SPE - $35/hour **
    • Nikon A1R - $50/hour
    • TIRF - $40/hour *
    • Keyence - $15/hour
    • Imaris Software - $10/hour
    • Confocal Microscope Training - $150

    * For longer TIRF experiments, $40/hour for the first 8 hours and then $10/hour thereafter.

    ** Because the Leica SP8s are heavily booked, we are offering a reduced rate of $35/hour on the SPE to encourage its use.

  • This facility is equipped with three microscopes: the Zeiss Axioplan upright, the Nikon E800 upright, and the Nikon TE300 inverted. Both Nikon microscopes are connected to cameras and equipped with Nikon NIS Element software for image acquisition. The Zeiss Axioplan upright is primarily used to confirm immunoreaction before image acquisition.

    The facility has an automated stitching epifluorescence system, the Keyence, which is capable of imaging 4 channels (excitation at 405nm, 488nm, 561nm, 647nm) as well as brightfield and can scan three slides at a time. We also have three standard epifluorescent microscopes: the Zeiss Axioplan upright, the Nikon E800 upright, and the Nikon TE300 inverted. Both Nikon microscopes are connected to cameras and equipped with Nikon NIS Element software for image acquisition. The Zeiss Axioplan upright is primarily used to confirm immunoreaction before image acquisition.

      Keyence Zeiss Axioplan Nikon E800 Nikon TE300
    Configuration Inverted Upright Upright Inverted
    Objectives        
         2x X   X  
         4x/5x X X X  
         10x X X X X
         20x X X X X
         40x   X    
         40x oil X   X X
         60x oil X X X  
    Fluorophores        
         DAPI X X X  
         CFP       X
         GFP/Alexa 488 X X X X
         YFP X   X  
         Cy3/RFP/Texas Red   X X  
         Texas Red/mCherry       X
         Cy5/Alexa 647 X   X  
  • Four confocal microscopes are available: Leica SPE, Nikon A1R, standard Leica SP8, and Leica Falcon SP8 with a high-speed resonant scanner, fluorescence lifetime imaging (FLIM) capability and deconvolution software.

    The Leica SP8s are configured with inverted microscopes, have automated stages and auto-focus, have stitching capability and are equipped with stage-top incubators for live cell imaging. Both instruments have Leica Navigator software. Excitation lines are as shown in the table below. The Leica SPE instrument is configured with an upright microscope. All Leica confocal microscopes are equipped with a spectral scanner system providing a spectral range of detection from 400 to 850 nm.

    The Nikon A1R confocal employs an inverted microscope with an automated stage and the Perfect Focus mechanism for time-lapse imaging or stitching of multiple image fields. It is also equipped with a stage top incubator. There are two scanheads available, a point scanner (conventional) and a resonant scanner. Simultaneous photoactivation/bleaching (using the 405nm laser) and imaging can be performed. Excitation lines are shown in the table below.

      Leica SP8  Leica SPE Nikon A1R Nikon TIRF
    Excitation        
         405nm x x x  
         440nm x     x
         457nm x   x  
         488nm x x x x
         514nm x   x x
         561nm x x x x
         594 nm x      
         639nm x x x x
    Brightfield x x x x
    Live Imaging x   x x
    Water Immersion Lenses   x    
  • The facility offers the use of Imaris for 3D image analysis, as well as Metamorph and NIS Elements for image quantification. Imaris is located in Arnold 309 and NIS Elements and Metamorph are installed on the Imaging Workstation in Stearns 207.

  • TIRF (total internal reflection flurorescence) microscopy produces very thin optical sections by creating an evanescent wave of excitation just 100nm above the coverslip without exciting molecules deeper within the specimen.  This results in optical sections much thinner than those obtained by the confocal microscope, but specimens must be within the evanescent excitation wave and must be mounted in aqueous media.  This inverted microscope is equipped with DIC and Perfect Focus, and does have a stage top incubator for live cell studies.  There is a choice of two cameras, an EM-CCD and an ORCA-ER CCD. 

    The laser excitation lines available include 442nm, 488nm, 514nm, 561nm, and 639nm. 

    Single channel imaging filter sets are available for 488 nm, 561 nm, and 639 nm excitable fluorochromes, and almost simultaneous dual channel imaging for 488 /561 nm is possible. A quadruple filter cube (442/514/561/639nm) can be used for CFP/YFP/RFP/far red imaging.