The present invention discloses and claims a quantitative brightfield image restoration (QRBF) method that faithfully recovers the shape and allows quantification of the size of individual unstained samples. The QRBF method restores out-of-focus brightfield images of unstained samples by applying deconvolution, which enhances contrast and enables quantitative analysis. To perform deconvolution, our procedure uses a point spread function (PSF) modeled from theory. Image deconvolution can be performed even from a single two-dimensional input image ("2D-QRBF"), and quantitative information such as sample size and shape can be determined from the 2D-QRBF-restored images. The application of 2D-QRBF in a high-throughput sampling process to assess cell shape changes during hyperosmotic shock shows that the described digital restoration approach is suitable for quantitative analysis of unstained brightfield images in a high-throughput imaging cytometry process.
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