Fluorescence Microscopy/Image Analysis Center (FMIAC)

The FMIAC is housed in MRB. It consists of equipment for obtaining digitized images of gels, blots, and microtiter plates for radioactivity, chemiluminescence and chemifluorescence in the Macroscopic Imaging Facility and for various light/fluorescence microscopy applications in the Microscopic Imaging facility.

Macroscopic Imaging Facilities

fmiac_1Macroscopic Imaging Facility Supervisor: Holly Giebler, Research Associate; Tel. 1-5017

The macroscopic imaging equipment in room 216 MRB includes a Typhoon FLA 9500 for radioisotopic, chemiluminescence and fluorescence imaging. Laser lines of 473, 532, 635, 685 and 785 are available. Large format phosphorimager screens allow detection of radioactivity over a 105 dynamic range.  An ImageQuant LAS 500 detection system for chemiluminescence, fluorescence and colorometric measurements from microtiter format plates is available in the same room. Across the hall in room 214 MRB is a LiCor Odyssey CLx near IR Scanner for dual color Western blot analysis. This equipment is not reserved but open to users on a first-come first-served basis. Users from outside the department need to be trained and have an account on file for hourly charges incurred.

Fees For Macroscopic Imaging Equipment: Odyssey costs are billed at $235/month divided by hours of use, averaged over 6 months. Typical hourly costs are between $25-30. Typhoon costs are billed at $450/month divided by hours of use, averaged over two month periods. Typical hourly costs range from $30-70.

 

Microscopic Imaging Facilities

Microscope Imaging Facility Supervisors: Olympus Spinning Disk Microscope: Dr. Barbara Bernstein, Senior Research Scientist, Tel: 1-0430; Nikon TIRF Microscope: Keith DeLuca, Research Associate, Tel: 1-5149; Nikon Diaphots, Laurie Minamide, Senior Research Associate, Tel: 1-5531.

The Microscope Imaging Center is housed in room 224 MRB. The fluorescence microscopes in this center are part of the CSU Microscope Imaging Network.

fmiac_2Nikon Total Internal Reflection Fluorescence (TIRF) Inverted Microscope (Eclipse Ti) has perfect focus control, a four laser line launch (405, 488, 561 and 640 nm) and an Intensilight Hg illumination source. A motorized XY Piezo Z stage and stage-top environmental chamber with temperature, CO2/air, humidity capability, will support culture dishes, multi-well plates or slides. Equipped with 20X, 40X, 60X DIC objectives and a 100X TIRF objective. Microscope has an Andor Clara camera for wide field imaging and an Andor iXon3 EMCCD camera for TIRF imaging. Microscope has built-in N-STORM super-resolution imaging capability and is networked to a RAID array for multiple terabyte storage. Nikon elements software controls all aspects of acquisition and analysis with a deconvolution software package included.

fmiac_3Olympus IX81 Inverted Spinning Disk Microscope has environmental control chamber with CO2 control for live cell work, an X,Y, piezo Z stage for rapid image stack acquisition across many predetermined fields (4 D imaging), a CSU 22 head with quad dichroic and additional emission filter wheel to eliminate spectral crossover, four high power diode lasers (405 nm, 488 nm, 561 nm and 647 nm) with rapid (microsecond) switcher and a phasor holographic photobleaching/photoactivation/photoconversion system for intracellular molecular dynamic measurements. Phasor can also be used for ion uncaging and other applications. System has differential interference contrast (DIC) optics with 10, 20, 40, 60 and 100X objectives, built in correction for spherical aberration for all objectives, and a wide field Xenon light source.  A cascade II EMCCD camera (confocal imaging) and a Photometrics HQ camera (wide field imaging) are both integrated for image capture using Slidebook software.  Laser selection and power are software controlled.  System is connected to RAID terabyte storage. The microscope is also equipped with a mosaic digital diaphragm system that uses thousands of adjustable mirrors to reflect a light beam simultaneously onto single or multiple user selected regions of interest on the image. These regions are identified by drawing around the spots of interest on the image screen.  A mercury light source is currently the only illumination system available for the mosaic.

fmiac_4Nikon Diaphot inverted epifluorescence microscopes (two) with video cameras for digital imaging are also available for routine fluorescence imaging on fixed samples. Filter cubes for all common fluors (DAPI, Fluorescein, Rhodamine, Texas Red, Cy 3, Cy5) are available and are interchangeable between microscopes. One microscope has DIC optics, an optional phase oil-immersion condenser, and a stage incubator and filter wheel for live cell studies (ratio imaging included) and one has a color camera for imaging stained sections. A specialized light source for darkfield illumination of slides is available and is especially useful for counting silver grains on emulsion treated samples. All microscopes utilize Metamorph software for image capture and analysis.

Data Storage
Users must bring their own drives for downloading image files as soon as the experiments are complete. Storage capacity for a longer time may be purchased on the Terabyte drive system.

Fees For Microscope Equipment: Use of Nikon Diaphot Microscopes is billed at $3.50 per hour (min use is 1 h).  Spinning disk confocal microscope and TIRF microscope have two methods of billing:. Infrequent users can pay hourly costs from the following scale: 1 h or less: $40/h; Second hour: $20; Third hour: $10; Fourth and additional hours: $5 to a maximum charge of $120 per 24 h period. Frequent users can pay an annual upfront charge of $2,500 for “unlimited use.” For frequent users starting after August 1, charges will be prorated at $210 per month with annual billing due in advance on or before July 1 for the following year.  Short term projects will be billed at $400 per month.

 


 

Schedule Time: Confocal

Access data on NAS1: Web  Mac OS X  On Windows, map network drive to \\fmic-nas1.bmb.colostate.edu\home

E-mail Userlist: Confocal