A three-dimensional localization and reconstruction method of multiple quantum dots in cells

Abstract

An intracellular multi-quantum-dot three-dimensional localization and reconstruction method comprises the following steps: firstly irradiating multiple quantum dots in cells by laser, performing amplification and imaging on fluorescent lights, emitted by the quantum dots after being stimulated, by a microscope objective, after the fluorescent lights pass an optical filter, forming parallel lightsvia a first convex lens, then modulating to form a double helical point spread function via a phase modulator preset with a specific Gaussian-Laguerre mode modulation function, focusing via a second convex lens, and then enabling the focused lights to enter into an EMCCD detector to generate a helical double-light-spot image; adjusting an axial position of an objective table by controlling a displacement table to go up and down, and repeating, so that three-dimensional information of the quantum dots on different focusing planes can be detected; storing images obtained by the EMCCD detector ina work station, and firstly forming three-dimensional relative coordinates of the quantum dots; then establishing an absolute rectangular coordinate system, and converting relative coordinate valuesinto absolute coordinate values; and performing reconstruction on a quantum dot three-dimensional space model, thus obtaining a three-dimensional image of the quantum dots in the cell. The method provided by the invention realizes real-time and nondestructive intracellular quantum dot spatial distribution measurement.

Description

technical field [0001] The invention relates to the technical field of optical three-dimensional positioning, in particular to a method for three-dimensional positioning and reconstruction of multi-quantum dots in cells. Background technique [0002] In order to gain an in-depth understanding of the morphological changes of cells under physiological activities, reveal the process of cell attachment, growth, division, and withering from the morphological point of view, or study the transfer of key metabolic substances in cells, fluorescent probes are often used on the cell surface and The tissues and organs in the cells are marked, and then optical imaging methods are used to locate or track these probes in three-dimensional space. In this way, researchers can obtain the spatial distribution or movement of probes inside the cell, thereby providing a theoretical basis for the study of cell physiological activities, tissue morphological changes, and key metabolic substance traj...

AI Technical Summary

Problems solved by technology

However, its disadvantage is that only one object can be tracked at a time. When the object is multiple targets, it is necessary to adopt a new three-dimensional positioning method

[0004] Existing technology cannot realize real-time and non-destructive measurement of the spatial distribution of quantum dots in cells

Images