Abstract
Gradient images can be obtained using a rotating square mask to filter the angular spectra of the wavefront generated by a complex transmittance object. This method can be applied to measure the three-dimensional structure of microscopic biological samples through the relationship of the phase with the optical path length. This work describes the implementation of a system using an inverted optical microscope and shows the experimental results of phase maps generated by boar sperm cells.
Highlights
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In the same cited work, we presented a procedure to correct the sensor characteristic nonlinear gain using calibrated polystyrene microspheres and obtaining a response matrix that incorporates the effect of the structure of the illumination source
We extend our previous results by showing the practical application of the Fourier plane rotating mask method in quantitative phase microscopy of biological samples with sub-cellular resolution
Summary
We extend our previous results by showing the practical application of the Fourier plane rotating mask method in quantitative phase microscopy of biological samples with sub-cellular resolution. The measurement is sequential, meaning one image is captured for each quadrant In this way, the intensity at a given point of the conjugate plane represents the corresponding quadrant intensity of the local wave vector at this point. As happens in digital holographic microscopy, the correct focus plane is not obvious from a simple camera image before processing [11] To manage this issue for posterior comparison, a nanopositioner is installed to hold the sample for registering phase maps with small variations in the distance between the sample and the microscope objective
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