Multi-photon exited near-infrared II fluorescence lifetime micro-imaging system

A fluorescence lifetime and microscopic imaging technology, which is applied in the directions of fluorescence/phosphorescence, material excitation analysis, material analysis through optical means, etc. Informative and easy-to-use effects

CN108982445AInactive Publication Date: 2018-12-11ZHEJIANG UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2018-12-11
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a multi-photon exited near-infrared II fluorescence lifetime micro-imaging system. In the invention, a 1220 nm femtosecond laser is introduced into an Olympus upright scanningmicroscope, and focused on a sample through a near-infrared anti-reflection objective lens; a near-infrared II fluorescence signal is collected by a large-diameter fiber collimator and coupled into anear-infrared anti-reflection large-diameter optical fiber, and finally detected and converted into an electrical signal by a photomultiplier tube responded by near-infrared II; the electrical signalis amplified and converted into an electrical counting signal by a signal amplifier and input to a TCSPC (time-correlated single photon counting) board, and at the same time the TCSPC counting board receives a synchronous pulse signal, as a timing stopping signal, from a laser for calculate time information of received photons; and after processing by a computer, a near-infrared II two-photon fluorescence lifetime image and a near-infrared II two-photon fluorescence intensity image are achieved. The invention is simple, practical, stable and flexible.
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Description

technical field

[0001] The invention belongs to the field of microscopic imaging of applied optics, and relates to a near-infrared two-zone fluorescence lifetime microscopic imaging system excited by multiphotons. Background technique

[0002] 1. Fluorescence microscopy imaging in the second near-infrared region excited by multiphoton

[0003] According to the relevant theory of the biological tissue window, fluorescence in the second near-infrared region (1000-1700nm) has a strong penetrating ability in biological tissue, which is conducive to the realization of large-depth imaging, and biological tissue does not have autofluorescence in this band, so The image has a high signal-to-noise ratio. At present, there are commercial near-infrared second-region fluorescence macroscopic imaging systems on the market, which can realize near-infrared second-region fluorescence macroscopic (whole body) imaging of biological specimens (such as mice). However, this imaging method has ...

Examples

Embodiment Construction

[0023] The present invention will be further described below in conjunction with accompanying drawing.

[0024] The present invention builds a set of multi-photon excited near-infrared second region (1000-1700nm) fluorescence lifetime microscopy imaging system. It is based on a scanning microscope, using a femtosecond pulsed laser with tunable wavelength as a light source (here 1220nm femtosecond laser is taken as an example to test the effectiveness of the system), through a near-infrared anti-reflection large-diameter fiber collimator and Near-infrared anti-reflective large-caliber optical fiber introduces the near-infrared second region fluorescence of the sample into the near-infrared second region response photomultiplier tube (PMT) (H12397-75) and converts it into an electrical signal, and cooperates with Becker&Hickl's counting board (SPC- 150), using TCSPC technology to realize multiphoton fluorescence lifetime microscopy imaging in the second near-infrared region.

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