Surface plasmon coupled emission directional enhanced microscopic fluorescence imaging and spectral detection method and device based on objective lens coupling

Abstract

The invention discloses a surface plasmon coupled emission (SPCE) directional enhanced microscopic fluorescence imaging and spectral detection method and device based on an objective lens coupling type. The polarized excitation light modulated by a half-wave plate is reflected by a dichroic filter to Arriving at the objective lens at a specific angle, exciting the metal surface plasmon to couple with the sample, the generated SPCE signal is collected by the objective lens and filtered by a dichroic filter, and then imported into the SPCE microfluorescence imaging module or SPCE microfluorescence spectrum through the mirror switch The module realizes the acquisition of SPCE microscopic fluorescence imaging and spectrum of the same micro-area of ​​the sample. In the present invention, by combining the microscope system, the coupling strategy SPCE is applied to high-spatial-resolution fluorescence imaging and spectroscopy at the same time. It has directional enhancement characteristics and a spatial selection function on the micro-area scale, and can obtain high-quality micro-fluorescence imaging and spectroscopy of the same micro-area of ​​the sample. Spectral information provides an efficient and reliable means for detecting low-content components in micro-regions of interest in complex systems.

Description

technical field [0001] The invention relates to the technical field of microscopic optical imaging and spectral analysis, in particular to a method and device for directional enhanced microscopic fluorescence imaging and spectral detection based on surface plasmon coupled emission (SPCE). Background technique [0002] With the continuous development of cells and small molecules in the fields of biology and medicine, it also puts forward higher requirements for microscopic analysis. The microscopic imaging and spectroscopy device is a fusion of optical imaging technology and spectroscopy technology. It can not only provide spatial imaging information of the target object, but also provide rich spectral information, providing a new technical means for studying or observing tiny objects. Fluorescence technology has the advantages of fast imaging speed, good repeatability, and intuitive observation, and has been more and more widely used in the fields of cells and small molecule...

AI Technical Summary

Problems solved by technology

At present, although the SPCE imaging and spectral detection device based on the prism configuration can obtain the SPCE fluorescence imaging and spectral information of the sample at the same time, its detection range is large, and it cannot detect samples with a micro-area size, which limits the application of SPCE analysis technology in more Applications on a Small Scale

On the other hand, although the SPCE technology based on the configuration of the objective lens can obtain the imaging information of the micro-scale sample, it is still unable to obtain the spectral information of the corresponding micro-area of ​​the sample.

Images