Multi-point unmarked differential super-resolution imaging method and device

Abstract

The invention discloses a multi-focus unmarked differential super-resolution imaging method and device based on an area array detector and Airy spot subdivision, light emitted by a laser is divided by a polarizing beam splitter into two beams of light with mutually vertical polarization directions, and the two beams of light are modulated by phase masks loaded by a left half screen and a right half screen of an SLM respectively; the two light beams are a solid light beam and a hollow light beam respectively; and then the solid light beam and the hollow light beam are combined, the combined light beam is divided into a first sub-light beam and a second sub-light beam, the first sub-light beam and the second sub-light beam respectively comprise the solid light beam and the hollow light beam, the first sub-light beam and the second sub-light beam enter a scanning galvanometer module at a certain angle and are focused by an objective lens to form a first focal spot combination and a second focal spot combination, and therefore four focal spots are formed on a focal plane. Based on a method of converting a time domain into a space domain, a single-point detector is replaced by an area array detector, and subdivision of more than 40 detectors can be carried out on Airy spot 4 at relatively low cost. And meanwhile, multi-focus excitation is adopted, so that the imaging efficiency of the system is further improved.

Description

technical field [0001] The invention belongs to the field of optical engineering, in particular to a multi-point non-marking differential super-resolution imaging method and device based on an area array detector and Airy disc subdivision. Background technique [0002] Optical super-resolution imaging has the great advantages of non-destructive, intuitive, and high-resolution. In the past two decades, a variety of super-resolution microscopes have been proposed and developed. These techniques can be divided into two categories: ① fluorescent labeling super-resolution microscopy, including emission loss microscopy, localization microscopy, structured illumination microscopy, etc.; ② non-labeling super-resolution microscopy. Label-free microscopy can visually observe the sample structure without being limited by dyes. However, compared with fluorescence imaging techniques, label-free microscopy develops relatively slowly in terms of super-resolution capabilities. Currently, t...

AI Technical Summary

Problems solved by technology

In the differential imaging technique reported so far, up to 19 single photon counters are used, which is limited by the system cost

The relevant confocal equipment of Zeiss can realize the subdivision of 37 detectors, but the detectors cannot be sold separately, and are currently only used for common confocal imaging

More subdivisions can be achieved by using an area array detector, but because the single-frame exposure speed of the area array detector cannot be compared with that of the single photon detector, the imaging speed is greatly reduced

[0004] In addition, traditional differential imaging needs to scan the sample twice with a hollow focal spot and a solid focal spot, and the imaging speed is twice as slow as confocal under the same conditions

Some researchers use dual focal points (hollow focal spot and solid focal spot) to scan in parallel and detect at the same time to achieve differential imaging, but the system structure is relatively complicated and two single photon counters are required

Images