New use of neodymium ion sensitized up-conversion nanocrystal, and high-resolution multi-photon microscopic system

Abstract

The invention discloses a new use of a neodymium ion sensitized up-conversion nanocrystal, and a high-resolution multi-photon microscopic system. The above neodymium ion sensitized up-conversion nanomaterial can be excited by short-wavelength stable laser with the center wavelength being shorter than 800nm to generate multi-photon visible light, so the nanomaterial has large multi-photon absorption cross section and multi-photon saturated excitation power, and makes higher-order multi-photon imaging easy, so the above new characteristic can be used in multi-photon microscopic imaging to greatly reduce the cost of the system and greatly improves the microscopic imaging resolution. The multi-photon microscopic imaging system comprises a short-wavelength stable laser with the center wavelength being shorter than 800nm, and the neodymium ion sensitized up-conversion nanomaterial is adopted as a sample. The short-wavelength stable laser is used to construct the cheap and simple multi-photon microscopic system for the first time, the neodymium ion sensitized up-conversion luminescence nanomaterial is used to carry out ultrahigh-resolution multi-photon microscopic imaging, and the material can also be introduced to cells, tissues or other matrixes in order to carry out high-resolution biological imaging.

Description

technical field [0001] The invention belongs to the technical field of optical microscopy, and in particular relates to a new application of neodymium ion-sensitized up-conversion nanocrystals and a high-resolution multiphoton microscopy system. Background technique [0002] In modern optical imaging techniques, probe-labeled fluorescence microscopy, especially laser scanning confocal microscopy based on fluorescence single-photon processes, has been achieved in the fields of medicine, life sciences, and materials science due to its high resolution (about 200 nanometers). Wide range of applications. However, confocal microscopy also has many shortcomings, such as relatively complex system, ultraviolet and blue light excitation is easy to damage biological samples, the imaging depth is very limited (tens of microns), it is difficult to avoid the interference of biological sample autofluorescence, and samples are easy to photobleach, etc. Based on the natural "optical section...

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Problems solved by technology

[0003] However, the nonlinear multiphoton (two-photon, three-photon, four-photon) excitation cross section of most fluorescent substances is very small, and the use of ultra-high peak power femtosecond pulsed lasers can obtain stronger multiphoton fluorescence, but the high cost of light sources ( ~$200,000) limits its widespread promotion, and the use of three-photon and four-photon fluorescence wavelengths under short-wavelength excitation usually falls in the ultraviolet band, which requires high detection optical paths and detectors, which is not conducive to short-wavelength excitation. Multiphoton High Resolution Microscopy

Although low-cost two-photon microscopy with steady-state laser pumping has been proposed, the resolution of low-order two-photon imaging excited by near-infrared wavelengths (greater than 800 nm) is still not high

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