CN1166939C - Multi-photon stimualting fluorescence method based on Q-regulating pulse laser - Google Patents
Multi-photon stimualting fluorescence method based on Q-regulating pulse laser Download PDFInfo
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- CN1166939C CN1166939C CNB011066539A CN01106653A CN1166939C CN 1166939 C CN1166939 C CN 1166939C CN B011066539 A CNB011066539 A CN B011066539A CN 01106653 A CN01106653 A CN 01106653A CN 1166939 C CN1166939 C CN 1166939C
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- pulse laser
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- excited fluorescence
- fluorescence combined
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Abstract
The present invention belongs to a multi-photon excitation method based on Q-switching nanosecond pulse lasers. A Q-switching technique and multi-photon excitation are combined to realize multi-photon excitation based on Q-switching pulse laser. The multi-photon excitation method based on Q-switching nanosecond pulse lasers comprises the following steps: 1), obtaining pulse laser with high-peak power by the Q-switching technique, wherein pulse width is in nanosecond magnitude; 2), exciting biologic samples which are labeled by a fluorescent probe or can generate autofluorescence by the Q-switching pulse laser for producing multi-photon excitation fluorescence. The present invention can be used for the research of subjects, such as biology, medical science, etc.
Description
Technical field
The invention belongs to a kind of a kind of method that realizes Excited Fluorescence Combined based on adjusting Q pulse laser.Can be used for the research of subjects such as biological and medical science.
Background technology
Because phototoxicity is little, advantages such as the good and high resolving power of penetrability, Excited Fluorescence Combined microscopic system (MPE) has become the important tool of biomedical research.
Multiphoton excitation is a kind of technology that has been widely used, and especially in the research in fields such as biomedicine, multiphoton excitation has become a kind of requisite technological means.Applied for multinomial patent abroad in this respect, for example U.S. Patent No. 5523573, U.S. Patent No. 5777732, U.S. Patent No. 5034613, U.S. Patent No. 5952192, U.S. Patent No. 6154310.In these patents or use locked mode ultrashort (psec or femtosecond) pulse laser perhaps to use continuous laser as excitation source as excitation source.Locked mode ultrashort (psec or femtosecond) pulsed laser technical operation is all very complicated, and this brings very big difficulty for user of non-optical specialty, and locked mode ultrashort (psec or femtosecond) pulsed laser especially the femtosecond pulse price is very expensive.Continuous light needs the very high average power that excites as the multiphoton excitation source, the feasible Excited Fluorescence Combined activating system based on continuous light of the high scattering of high-average power and high thermal absorption is difficult to become utility system, and does not have the report of continuous light three-photon fluorescence excitation so far.
Summary of the invention
The objective of the invention is to propose a kind of Excited Fluorescence Combined method based on adjusting Q pulse laser at the problem and shortage that has now based on the Excited Fluorescence Combined system of ultra-short pulse laser or continuous laser.
For when can obtaining high fluorescent, do not produce tangible thermal effect again and reduce the influence of light scattering measuring, exciting light must have the characteristics of high-peak power and low average power, this just requires pulsed laser to have low-repetition-frequency, and Q-switched pulse laser obviously is best selection.
The said Excited Fluorescence Combined method principle based on adjusting Q pulse laser of the present invention is, transit to excited state after making sample molecule (or group) absorb two or more photons simultaneously, ground state and emitting fluorescence (or phosphorescence) are got back in transition then, and exciting light adopts adjusting Q pulse laser.
The step that realizes Excited Fluorescence Combined is: 1) adopt Q-regulating technique to obtain the pulse laser of high-peak power, pulse width is a nanosecond order; 2) utilize adjusting Q pulse laser to excite the biological sample that maybe can produce autofluorescence, produce Excited Fluorescence Combined with the fluorescence probe mark.
In the said Excited Fluorescence Combined method based on adjusting Q pulse laser of the present invention, its Q-switched laser is simple in structure, and price is lower, stable performance, and the beam quality height, and also easy to operate.The pulsewidth of adjusting Q pulse laser is a nanosecond order or wideer, and its repetition frequency is easy to regulate.Compare with the high repetition frequency of mode-locked laser, utilize the low-repetition-frequency characteristic of Q-switched laser can obtain the pulse laser of high-peak power and low average power.This just multiphoton excitation to study the multiphoton excitation research of biological living especially necessary.
Embodiment
In the said Excited Fluorescence Combined method based on adjusting Q pulse laser of the present invention, said excitation pulse laser is produced by Q-switched pulse laser.
Said Q-switched pulse laser can be a solid state laser, also can dye laser.YAG laser instrument for example, laser instruments such as ti sapphire laser and rhodamine laser instrument.
The pulsewidth of said Q-switched pulse laser is a nanosecond order or wideer.
The repetition frequency of said Q-switched pulse laser can be regulated from 1Hz to 1MHz.
Claims (7)
1. Excited Fluorescence Combined method based on adjusting Q pulse laser, its step that realizes Excited Fluorescence Combined is: 1) adopt Q-regulating technique to obtain the pulse laser of high-peak power; 2) utilize adjusting Q pulse laser to excite object, produce Excited Fluorescence Combined.
2. according to the said Excited Fluorescence Combined method of claim 1, it is characterized in that excitation pulse laser is produced by Q-switched pulse laser.
3. according to the said Excited Fluorescence Combined method of claim 1, it is characterized in that said Q-switched laser is a solid state laser.
4. according to the said Excited Fluorescence Combined method of claim 1, it is characterized in that said Q-switched laser is a dye laser.
5. according to the said Excited Fluorescence Combined method of claim 1, it is characterized in that the pulsewidth of said Q-switched pulse laser is a nanosecond order.
6. according to the said Excited Fluorescence Combined method of claim 1, it is characterized in that the repetition frequency of said Q-switched pulse laser can be regulated from 1Hz to 1MHz.
7. according to the said Excited Fluorescence Combined method of claim 1, it is characterized in that multiphoton excitation is to liking the biological sample that maybe can produce autofluorescence with the fluorescence probe mark.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011066539A CN1166939C (en) | 2001-04-23 | 2001-04-23 | Multi-photon stimualting fluorescence method based on Q-regulating pulse laser |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011066539A CN1166939C (en) | 2001-04-23 | 2001-04-23 | Multi-photon stimualting fluorescence method based on Q-regulating pulse laser |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1317688A CN1317688A (en) | 2001-10-17 |
| CN1166939C true CN1166939C (en) | 2004-09-15 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011066539A Expired - Fee Related CN1166939C (en) | 2001-04-23 | 2001-04-23 | Multi-photon stimualting fluorescence method based on Q-regulating pulse laser |
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| CN (1) | CN1166939C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6075963B2 (en) * | 2012-03-26 | 2017-02-08 | オリンパス株式会社 | Fluorescence observation method and fluorescence observation apparatus |
| WO2017012555A1 (en) * | 2015-07-20 | 2017-01-26 | 徐敏 | Photon structure and chemometrics pathologic system |
| CN106568754A (en) * | 2016-11-06 | 2017-04-19 | 浙江大学 | Optical system used for measuring liquid sample multiphoton fluorescence spectrum |
| CN108051413A (en) * | 2017-11-30 | 2018-05-18 | 百色学院 | A kind of light activated photoluminescence spectra measuring system of pulse |
| CN111537444B (en) * | 2020-05-25 | 2021-07-06 | 南京航空航天大学 | Laser ultrasonic nondestructive testing method and system with repetition frequency virtually regulated |
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2001
- 2001-04-23 CN CNB011066539A patent/CN1166939C/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| CN1317688A (en) | 2001-10-17 |
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