Laser scanning phase microimaging method and system

Abstract

The invention relates to laser scanning phase microimaging method and system. The system comprises a laser confocal fluorescence scanning microscope system consisting of a laser device, a dichroscope, an objective lens, a scanning platform, a convergent lens, a pinhole and a photoelectric detector and a phase detection system consisting of a laser transmitting component and a fluorescence shielding component, wherein the laser ejected by the laser device is emitted to the dichroscope through the laser transmitting component for splitting; the ejected laser is focused on a sample on the scanning platform through the objective lens, the laser focused to the sample penetrates through the sample to be emitted to a fluorescence medium under the sample, a laser spot area is partially overlapped with an area capable of exciting a detectable fluorescence, and the overlapped area is excited by the laser to emit fluorescence; the fluorescence transmits over the sample to be emitted to the dichroscope through the objective lens for splitting, the ejected fluorescence is emitted to the photoelectric detector through the convergent lens and the pinhole to detect so as to obtain the phase of one measurement point of the sample; the scanning platform drives the sample and the fluorescence medium to move, and the phases of all the measurement points of the detected sample are processed to obtain a relief-like structural image of the sample. The method and the system disclosed by the invention can be widely applied to microimaging of a biological sample.

Description

technical field [0001] The invention relates to an optical imaging method and system, in particular to a laser scanning phase microscopic imaging method and system. Background technique [0002] Laser confocal fluorescence scanning microscopy is an imaging process that uses a pinhole placed in front of the detector to achieve high-quality point scanning and point detection of fluorescence. The contrast of laser confocal fluorescence scanning microscopy images comes from the fluorescence intensity generated by laser excitation of samples, so non-fluorescence labeled structure imaging of transparent samples cannot be achieved alone. [0003] In the prior art, the biological tissue microphase imaging method that can be combined with laser confocal fluorescence scanning microscopy mainly includes: 1. The laser oblique illumination microscopic imaging method is based on the confocal microscopy system, using incident laser light The angle formed by the optical axis and the detect...

AI Technical Summary

Problems solved by technology

However, due to the need to adjust the angle between the detection light path and the incident light path, it is difficult to achieve phase measurement without destroying the original confocal light path.

2. The differential interference phase contrast microscopy imaging method uses two Wollaston Prism, polarizer and analyzer to complete the imaging. The imaging result can present the relief-like structure of biological tissue, but The above microscopic imaging method needs to add a polarizing element in the optical path to generate polarized light, and use the shear angle formed by the two polarized lights to detect the phase difference. The polarizing element used is expensive and cannot detect birefringent samples. to detect

Images