Surface plasma coupling emission directional enhancement type microscopic fluorescence imaging and spectrum detection method and device based on objective lens coupling type

Abstract

The invention discloses a surface plasma coupled emission (SPCE) directional enhanced microscopic fluorescence imaging and spectrum detection method and device based on an objective lens coupling type. Polarized exciting light modulated by a half-wave plate is reflected by a dichroic filter and then reaches an objective lens at a specific angle to excite metal surface plasma to be coupled to a sample, and generated SPCE signals are collected by the objective lens and filtered by the dichroic filter and then are switched and guided into an SPCE microscopy fluorescence imaging module or an SPCE microscopy fluorescence spectrum module through a reflector to realize SPCE microscopic fluorescence imaging and spectrum acquisition of the same micro-area of the sample. By combining a microscope system, a coupling strategy SPCE is applied to high-spatial-resolution fluorescence imaging and spectrum at the same time, the directional enhancement characteristic and the spatial selection function on the micro-area scale are achieved, high-quality microscopic fluorescence imaging and spectrum information of the same micro-area of a sample can be obtained, and an efficient and reliable means is provided for detecting the low-content components of a micro area of interest in a complex system.

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...

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