The DeltaVision Restoration Microscope is an automated epifluorescence microscope coupled to a precision scanning stage. The resolution of stage movement is 10 nm in X and Y and 4 nm (that's not a typo) in Z. Interchangeable computer-controlled filter wheels provide nearly limitless options for imaging fluorescent probes. We have two sets, one for standard blue, green, red and far red dyes, and another, "Live Cell" set, with filters for CFP, GFP, YFP and RFP. The optical properties of the microscope objectives are carefully defined, and the resulting PSF's (point spread functions) are used in a deconvolution algorithm to re-assign light to it's original point source. The outcome is an image with an improved signal to noise ratio and apparent improved resolution. The two images in the banner above, an axonal buton labeled with GFP and Alexa 555, illustrate the result of this process. The system is best utilized for relatively thin specimens (> 100 um), but is highly dependent on the inherent light scattering properties of the specimen. Thick, or highly diffusing specimens would be better imaged on the confocal microscope. The system includes an argon (488 nm) laser for carrying out FRAP (fluorescent recovery after photobleaching) experiments. Single spot sizes from 0.5 to 4 um can be selected with a beam expander. Bleaching larger areas (lines, regions of interest) is also possible. Software for image acquisition, colocalization analysis and FRAP analysis is included on the computer that controls the microscope. A stand-alone computer, linked to the system computer via FTP, has image viewing and 3-D rendering software (Imaris). For large files or multiple image sets, it is recommended that you use the stand-alone system to free the microscope for image acquisition. An initial 2 hour training session will introduce the user to the functions and proper operation of the microscope. Additional assistance is provided until the user become self-sufficient. After the initial training period, use of the system is charged at $25/hr. There is no charge for post-processing time on the stand-alone computer. |