Live-Cell Imaging

Several systems in the Microscopy Facility are set up for live cell imaging.

1. Spinning disc confocal microscope.  This system is devoted exclusively to live cell / live tissue imaging.  Illumination sources include lasers (405 nm, multi-line Ar [458/488/514 nm], 561 nm, 635 nm), and metal halide.  Objective lenses include 10x, 20x, 40x oil immersion, 63x oil immersion, 63x water immersion, 100x oil immersion.  The microscope also includes a full stage PeCon incubation system that regulates temperature, CO2, O2, and humidity.  Image detection is via 2 emCCD cameras.  A motorized XY stage permits multi-point time-lapse imaging.
2. Confocal laser scanning microscopes.  Two of the confocal laser scanning microscopes can be set up for live cell imaging.  A low-profile PeCon stage incubator (which regulates regulates temperature, CO2, and humidity) and objective heater are available to support live cell imaging.  Both systems (710 and SP2) have motorized XY stages that permit multi-point time-lapse imaging.
   1. The Zeiss 710 has 405, 440, 458,488, 514, 561 & 633 nm laser lines.  Objective lenses include 10x, 20x, 40x oil immersion, 63x oil immersion).  
   2. The Leica SP2 AOBS has 5 lasers (405 nm, multi-line Ar [458/488/514 nm], 543, 594, and 633 nm).  Objective lenses include 10x, 20x, 20x oil immersion, 40x oil immersion, 63x oil immersion).
    
3. Total Internal Reflection Fluorescence (TIRF) microscope.  Illumination sources include lasers (405 nm, multi-line Ar [458/488/514 nm], 561 nm) and metal halide.  Objective lenses include 10x, 20x, 40x, 63x oil immersion, 100x-TIRF oil immersion.  The microscope also includes a stage incubation system that regulates temperature, CO2, and humidity, an objective lens heater and Zero Drift Compensation autofocus.  Image detection is via an emCCD camera.
4. Multi-photon laser scanning microscope.  Although configured around an upright stand, the system has a stage heater and closed chamber system to permit live cell imaging as well as a heated stage to permit imaging of vital tissue slices.  Objective lenses include 10x, 20x, 25x, 40x oil immersion, 63x oil immersion, 40x water immersion as well as 20x, 40x and 63x dipping lenses for intravital imaging.
5. Structured Illumination microscope (SIM). The Nikon N-SIM structured illumination microscope can generate 'super-resolution' iimages of fluorescently labeled live cells in culture (as well as fixed samples). Laser lines include 405, 488, 561, and 640 nm. Objective lenses include 10x, 20x, 40x, 60x water (1.27 n.a.) & 100x SR TIRF (1.49 n.a.) oil immersion. The microscope is equipped with Perfect Focus laser-based automated focus drift compensation and a Tokai Hit stage incubator (which regulates temperature, CO2 and humidity) to facilitate live cell imaging.
6. A Zeiss Axiovert 200 widefield fluorescence microscope is equipped with a Hamamastu ORCA ER CCD camera, Colibri LED illumination unit (blue, green, red), white light LED and a stage heater for short-duration live cell imaging.

A dedicated cell culture room (equipped with a laminar flow hood / biological safety cabinet and CO2 water-jacketed incubators) is on site to support live cell imaging.  Cell lines may be maintained on-site for long term live cell imaging studies.