As an important marker in disease diagnosis, red blood cell morphology measurement is necessary in biological and medical fields. However, traditional setups as microscopes and cytometers cannot provide enough quantitative information in morphology detections. In order to capture tiny variations of red blood cells affected by metal ions in external environment, quantitative interferometric microscopy is applied: combining with phase retrieval and cell recognition, cellular phases as well as additional quantitative cellular parameters can be acquired automatically and accurately. The research proves that quantitative interferometric microscopy can be potentially applied in cellular observations and measurements for both biological and medical applications.
KEYWORDS: Digital imaging, Live cell imaging, Numerical simulations, Microscopy, Real time imaging, Phase retrieval, Microscopes, Image retrieval, Phase imaging, Digital image correlation
As an ideal way for quantitative live cell imaging, dual view transport of intensity equation (TIE) method can provide both real time imaging, multi-mode observations, simple setup and large field of view (FoV). However, the image recorder installation error reduces the accuracy in both amplitude and phase retrievals, because of the inevitable FoV mismatch between the captured under- and over-focus intensities. In order to obtain higher accuracy amplitude and phase retrievals, the phase correlation based digital FoV correction is introduced into our method, rotation, scale and translation between the under- and over-focus images are compensated by the phase correlation based digital FoV correction. Measurements are implemented using standard sample detection and quantitative live cell imaging, proving that the proposed method can improve the accurate of the amplitude and phase computations.
KEYWORDS: Digital imaging, Live cell imaging, Digital image correlation, Numerical simulations, Real time imaging, Phase retrieval, Optical engineering, Image processing, Microscopy, Blood
Dual-view transport of intensity equation (TIE) method is an ideal way for quantitative live cell imaging as it has advantages such as real-time imaging, multimode observations, compact setup, and large field of view (FoV). However, due to the image recorder installation error, the inevitable FoV mismatch between the captured under- and over-focus intensities reduces the accuracy in both amplitude and phase retrievals. Here, to eliminate this undesired FoV mismatch, the phase correlation-based digital FoV correction is adopted to recognize and compensate the rotation, scale, and translation between the under- and over-focus images. Both the numerical simulations as well as the experiments in standard sample detection and quantitative live cell imaging prove that the digital FoV correction combined dual-view TIE method can maintain the consistence of the dual FoVs, thus guaranteeing the high-accurate amplitude and phase computations, proving the proposed method is a promising quantitative live cell imaging tool in various applications such as biological observations and medical diagnostics.
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