A microscopic imaging system for analysis of integrated light interference and scattering information and a method

Abstract

A microscopic imaging system for analysis of integrated light interference and scattering information is disclosed and is based on a coaxial Michelson interference light path and a Mirau interference device. A sample in at an objective table that is a half mirror. A reflected light beam generates interference on the Michelson interference light path through the Mirau interference device, an amplified interference image is obtained by a microscope, and phase information of the sample can be extracted from the image. At the same time, after the forward transmission light beam passing through the sample passes through the half mirror, forward small-angle and large-angle scattering are generated. The scattering image can be received by a forward CCD, forward scattering amplitude distribution can be obtained from the scattering image, and the size of the sample can be deconstructed by utilization of the scattering theory. Through integrated phase and scattering information, the system and the method can detect the morphological structure of the sample more accurately. The system and the method have important practical value in the field of microscopic morphological measurement of transparent objects, particularly in the fields of biological cell substructure morphological detection and application.

Description

technical field [0001] The invention belongs to the technical field of original state microscopic imaging of biological cells, and in particular relates to a microscopic imaging system for comprehensively receiving and analyzing interference information and scattering information. Background technique [0002] Although most cells are often considered to be colorless and transparent, strictly speaking, they cannot be completely transparent, that is, they cannot be pure phase objects. Therefore, when light passes through cells, the amplitude and phase of light will change. Changes, both the amplitude and the phase of the light passing through the cell contain information about the cell. Phase imaging detection technology obtains cell information from the phase modulation of cells to incident light waves, while light scattering detection technology obtains cell information from the amplitude modulation of cells to incident light. Therefore, theoretically speaking, phase imagin...

AI Technical Summary

Problems solved by technology

In 2006, the research group of Professor Gabriel Popescu proposed Diffraction Phase Microscopy (DPM), which combines the advantages of off-axis optical path and coaxial optical path: it has good stability and high acquisition rate, but due to the light source, the diffraction phase Microscopic technology (DPM) images have speckles, which will reduce the sensitivity of spatial phase and limit its application in the study of cell substructure. In 2011, the research group of Professor Gabriel Popescu also proposed spatial light interference technology. (SLIM) uses white light as the light source to overcome the speckle effect, but this technique requires the acquisition of four intensity images for each quantitative phase map

However, the techniques mentioned above are all for obtaining the interference information of the sample, and can only detect the transparent phase body sample, and there are still difficulties in detecting the substructure of the sample.

[0004]Light scattering detection technology generally does not require interference, and directly measures the scattering intensity distribution, scattering energy distribution or light intensity attenuation, etc., and the information it provides is the measurement volume. The average value of , averages the contribution of a single component to the scattering, and the sample substructure shape has a great influence on its scattering, so it has an important application in the measurement of cell size, such as the patented technology WO 2014 / 084957 A1 (Optical system, apparatus and method for flow-cytometry: the optical system, device and method of flow cytometry), however, this type of technology is only analyzed from the aspect of scattering for the quantitative measurement of cell morphology changes, and does not involve interference information

Images