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Data from brightfield imaging, phase contrast (interference image), as well as phase gradient magnitude and DIC images, are also obtained simultaneously along with phase and optical thickness.
Using this model, we can simulate the interference image quickly.
Nomarski differential interference image was also recorded.
Only light diffracted at sufficiently small structures can pass the phase plate and contribute to the interference image.
In contrast to conventional methods of optical microscopy based on the registration of light intensity distribution, the laser interference microscopy allows obtaining the phase distribution in the interference image.
In order to correlate the pixels in the interference image with those pixels in the fluorescence image collected with a CCD (Coolsnap, Princeton Instruments), a single affine transformation (consisting of a scaling and a rotation) was applied to the scan region and the edges of the transformed images were subsequently cropped by a few pixels to produce rectangular regions.
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As expected, there is a strong correlation between the presence of paxillin and actin in stress fibers (as identified with fluorescence imaging) and the interference images.
Quantitative phase imaging can measure structures using interference images analogous to those viewed with phase-contrast, differential interference contrast and digital holography [ 2].
Using the prototype, the pushbroom imaging experiments are carried out and the original interference images are obtained.
Processing interference images in the three different directions can realize the three dimensional measurement.
These three interference images show a good agreement with each other that demonstrate the correctness of the optical model.
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