Total internal reflection fluorescence microscopic imaging method and device

A technology of microscopic imaging and total internal reflection, which is applied in the field of total internal reflection fluorescence microscopic imaging, can solve the problem of space limitation for placing samples, and achieve the effect of high resolution, high resolution and high microscopic imaging.

Active Publication Date: 2013-12-11
BEIJING INFORMATION SCI & TECH UNIV
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Problems solved by technology

In terms of detection, it is not easily disturbed by the incident light signal, but the space for placing the sample is limited by the prism

Method used

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  • Total internal reflection fluorescence microscopic imaging method and device
  • Total internal reflection fluorescence microscopic imaging method and device
  • Total internal reflection fluorescence microscopic imaging method and device

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Embodiment Construction

[0016] In the present invention, by using an axisymmetrically polarized beam with spatial polarization changes, the amplitude, phase, and polarization distribution of the incident laser beam can be adjusted, the distribution of the evanescent field can be adjusted, and a smaller "fluorescent probe" can be obtained to improve the detection efficiency. Spatial resolution, realizing three-dimensional super-resolution focusing.

[0017] Axisymmetrically polarized beams are a class of vector beams with axisymmetric polarization distribution characteristics, and the axis of symmetry is the propagation axis of the beam. Any point on the cross-section of the beam (except the central point) is linearly polarized, and the polarization azimuth change along the circumferential direction satisfies the following relationship,

[0018] Φ(r,φ)=P×φ+φ 0 (P≠0)) (1)

[0019] Among them, P is called the polarization order, which means the period number of the polarization azimuth change when ...

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Abstract

The invention provides a total internal reflection fluorescence microscopic imaging method and device based on senior secondary axisymmetric polarized beams. The device comprises a laser device for emitting laser beams, a pinhole filter for performing spatial filtration on the laser beams emitted from the laser device, a collimation lens for collimating the laser beams subjected to spatial filtration into parallel light beams, a polarization conversion system for performing polarization state conversion on the parallel light beams to obtain the senior secondary axisymmetric polarized beams, a pupil filter and an annular diaphragm which are used for modulating the vibration amplitude and the phase of the obtained senior secondary axisymmetric polarized beams, a reflecting and focusing system for focusing the modulated axisymmetric polarized beams on an interface between glass and a sample to produce an evanescent field on the interface between the glass and the sample because of total reflection, a reflecting and filtering system for focusing a fluorescence signal emitted from the evanescent field on a pinhole array plate, a photoelectric detector for detecting the signal, and a signal analysis processing system.

Description

technical field [0001] The invention relates to a total internal reflection fluorescence microscopic imaging technology, in particular to a total internal reflection fluorescent microscopic imaging method and device based on a high-level subaxially symmetric polarized light beam. Background technique [0002] Total Internal Reflection Fluorescence Microscopy (TIRFM) is an emerging optical imaging technology in recent years, which uses the evanescent field generated by total internal reflection to illuminate the sample, resulting in optical The fluorophores within the thin layer are excited and the signal-to-noise ratio of fluorescence imaging is high. The imaging device of this method is simple, and it is very easy to combine with other imaging techniques and detection techniques. At present, the spatial resolution of 100nm or even lower has been successfully achieved, and it is widely used by biophysicists in single-molecule fluorescence imaging. [0003] The introduction...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G02B21/06
Inventor 周哲海祝连庆娄小平王君张荫民刘谦哲孟晓辰
Owner BEIJING INFORMATION SCI & TECH UNIV
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