Booking

Colorado State University Users: Only trained users are allowed access to calendars for booking time on any MOAI microscope. Information on how to book time on any MOAI microscope will be provided to users upon completion of their training.

Non-Colorado State University Users: Potential users from outside CSU should contact the facility supervisor listed for each instrument to get current rates of access for training and use or fee-for-service costs.

Data Storage

All users should bring their own drives for downloading image files as soon as the experiments are complete. Data stored on the acquisition computers should not be left for longer than one week. Storage capacity for a longer time may be purchased on either an interconnected RAID Terabyte drive system or on a university data storage computer. These should be arranged by the laboratory PI after the initial training of lab personnel.

Selecting a Microscope

The numbers and capabilities of light microscopes available within the MOAI are extensive so selecting the best instrument for your needs is not always simple. If you are just starting to utilize microscopy for a project, discussing where to start with a facility supervisor is usually a good step. For working with cells, a first step would be getting phase contrast, DIC, oblique illumination images or simple fluorescence images if immunofluorescence staining or expression of fluorescently tagged molecules are present. The Keyence microscope is often a good starting point because of its simplicity to use (totally mouse controlled for focus and image capture) and availability of brightfield, darkfield, oblique illumination and fluorescence (filter cubes for most common fluors available). Living cells can be imaged under controlled temperature and CO_2. Images can be captured over large X,Y areas and easily stitched into one image but also using a Z-stack in multiple planes with as little as 0.1 μm steps. Full focus (projection image) of a stack is easily made.

Moving into higher resolution/single molecule imaging is another step that will involve selection of a Total Internal Reflection Fluorescence (TIRF) or confocal microscope. The TIRF allows imaging of fluorescence membrane-associated molecules or submembrane components and images are not compromised by having the molecules expressed throughout the cell since only the cell region in contact with the substratum and slightly internal to this plane will receive excitatory photons. The type of confocal microscope for selection will depend on the type of sample (inverted versus upright configuration of microscope), the speed and frequency needed for image acquisition, the size of the area to be imaged, the detection sensitivity required, and the availability of the correct wavelengths of light/filters for excitation and emission.