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The following illustrates the technique of deconvolution and restoration microscopy as implemented on the API DeltaVision system.

This image was acquired from a DAPI-stained preparation of 293T cells, provided by Dr. Lubbertus Mulder at ADARC, acquired at 100X magnification.

The DeltaVision microscope is based upon a standard fluorescence microscope, with the addition of precision X, Y and Z stage control. After finding and focusing on a field of interest, the DeltaVision system is set up by the investigator to automatically acquire a number of images (image stack) while the system mechanically focuses through the cell at precise intervals. The following image is number 16 from an image stack of 20 images, each of which is an image taken as the focus was changed by 0.2um. (click on image to enlarge)

At this point the image-processing computer takes over, using a Point Spread Function (PSF) file as a reference to deblur or deconvolve the original image stack. The PSF is a file that characterizes the microscope’s optical characteristics using the same objective lens, coverslip, and immersion oil.

This operation generates another image stack of deconvolved images, of which this is again the 16th image from the stack of 20. Notice the increase in resolution obtained by the removal of blurring. (click on image to enlarge)

This image stack may be further computer processed to yield a 3-D image of the cells (click on image to enlarge).