| The phase recovery technique based on the intensity transport equation(TIE)is a non-interfering,non-iterative,simply equipped phase imaging method that is fast and stable in imaging and high-contrast phase detection.It has been widely approved in biomicroscopy imaging.The traditional TIE phase recovery technique needs to move the object plane or image plane three times to collect at least three optical intensity images,which are the over-focus,the in-focal and the under-focus intensity images.However,it does not applicable in real-time phase imaging.In order to achieve a high-precision and low-error real-time phase imaging method for live cells,we propose a digital field-of-view correction dual-view TIE phase imaging technology in this paper,which overcomes the disadvantage of real-time imaging in traditional TIE phase imaging technology and solves mismatch of the binocular field of view in phase imaging,and it also realizes the measurement of biological cells by non-interferometry quantitative phase microscopic real-time imaging.The specific research work and results are as follows:(1)The wave equation without time is obtained by deriving the optical wave transfer equation and the Helmholtz equation,then the basic light intensity transport equation can be obtained in optical wave paraxial approximation.During the traditional TIE phase recovery experiment,we found that the traditional TIE technology cannot fit in real-time phase imaging.So we propose the dual-view TIE phase recovery algorithm and use the fast Fourier transform method to solve the phase of dual-view TIE.Next,the phase simulation of red blood cells under different noise conditions is carried out.The high-precision phase recovery results show that the dual-view TIE technology is significant in real-time phase imaging.(2)Two image sensors with the same type and specifications were installed on the binocular tube of the optical microscope.At the same time,a 3mm C-mount brass spacer rings was attached to one of the eyepiece cylinders,and two images were simultaneously exposed.Thus the phase is calculated by dual-view TIE recovery algorithm.It has been found that there are rotation,scaling and translation errors between the images acquired by the binoculars through experiments,which result in incorrect phase results.In order to calibrate the registration error between the two fields of view,we propose a real-time phase imaging technique based on phase correlation.The calibration algorithm and dual-view TIE algorithm were used to measure the resolution plate,random phase plate and human blood cell by simulation and experiment.The standard results proved that the field correction algorithm can accurately correct the error in the fields of view and the digitally corrected dual-view TIE phase imaging technology can be applied to phase detection in live cell,which enrich the real-time phase imaging method of living cells.(3)Digital field of view correction dual-view TIE in real-time phase imaging technology can be directly applied to biological microscopic systems.In this paper,the corrected dual-view TIE phase imaging technique was used to measure a variety of cells in experiments.Adding ATP and glucose to BHK21 and F81 live cells,then we calculate cell phase distribution in real time.According to the time-correlation algorithm and root mean square phase principle,the frequency and amplitude of cell membrane fluctuation were compared;the cells were added with ATP show higher phase fluctuation,which strongly proves that ATP can promote the mechanical regulation of cell membrane,and the digital field of view correction combined dual-view TIE technology has great prospects in living cell phase imaging.The digitally corrected dual-view TIE phase imaging technique is proposed in this paper with obvious improvement of speed in real-time phase imaging technology.It has broken the limitations of traditional TIE technology and provides a new phase imaging method for biomedical cell research. |
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