CN109612970B - Method and device for enhancing fluorescence intensity of gold nanospheres - Google Patents
Method and device for enhancing fluorescence intensity of gold nanospheres Download PDFInfo
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- CN109612970B CN109612970B CN201811403930.6A CN201811403930A CN109612970B CN 109612970 B CN109612970 B CN 109612970B CN 201811403930 A CN201811403930 A CN 201811403930A CN 109612970 B CN109612970 B CN 109612970B
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
Abstract
The present invention belongs to the field of nanometer material and optics. A method for enhancing fluorescence intensity of gold nanospheres is characterized in that continuous laser is focused on agglomerated gold nanosphere particles (4) with the diameter of 150-170 nanometers to excite the agglomerated gold nanosphere particles (4) to obtain gold nanosphere fluorescence, the gold nanosphere fluorescence is enhanced along with the increase of laser irradiation time, the optimal enhancement effect is obtained when the laser irradiation time reaches 4.5 seconds, and then the gold nanosphere fluorescence is weakened along with the increase of the laser irradiation time. The invention also relates to a device for enhancing the fluorescence intensity of the agglomerated gold nanospheres through continuous laser irradiation. According to the invention, the agglomerated gold nanospheres are irradiated by near ultraviolet continuous laser, so that adjacent gold nanospheres generate a welding effect under the action of a photo-thermal effect, and further a strong local surface plasmon enhancement effect is generated, so that the fluorescence intensity of the gold nanospheres is enhanced by more than 150 times.
Description
Technical Field
The invention belongs to the field of nano materials and optics, and particularly relates to a method and a device for enhancing fluorescence intensity of agglomerated gold nanospheres through continuous laser irradiation.
Background
Noble metal nanoparticles, such as gold nanospheres, have wide applications in many fields due to their unique physicochemical properties, such as excellent photo-thermal properties, excellent stability, bio-nontoxicity, efficient catalytic properties, and biocompatibility. Particularly, the local surface plasmon enhancement effect of the gold nanoparticles enables the gold nanoparticles to greatly enhance the linear and nonlinear optical response of materials (usually about 10 nm) close to the gold nanoparticles, and the gold nanoparticles are currently applied to scientific research and daily production such as surface-enhanced Raman spectroscopy, two-photon fluorescence, frequency doubling and the like.
The gold nanoparticles also have single-photon and two-photon fluorescence emission characteristics, so that the gold nanoparticles have excellent application prospects in the aspects of biological imaging, medical diagnosis and treatment, preparation of photoelectric devices and the like. However, the extremely low fluorescence quantum yield of gold nanoparticles limits their wide application in various fields. In order to improve the quantum yield of the fluorescence emission of the gold nanoparticles, the most common method at present is agglomeration induction enhancement, namely, a local surface plasmon enhancement effect of the gold nanoparticles is enhanced by forming a gold nanoparticle dimer, and the fluorescence emission intensity is improved. Another method for enhancing the fluorescence of the gold nanoparticles is to enhance the electronic state of the gold nanoparticles, which occupies the highest molecular orbit, by adding other suitable noble metals, so that the visible optical transition is enhanced, and the quantum yield is improved. These methods all have certain limitations, such as dimer acquisition requires a large scientific instrument for precise operation, the cost is high, the operation is complex, and the method is difficult to be used in industrial production; the addition of other noble metals has a limited effect on enhancing the fluorescence of the gold nanoparticles, and the fluorescence intensity of the gold nanoparticles can be increased by only 20 times.
According to the invention, the agglomerated gold nanospheres are irradiated by continuous laser, so that the gold nanospheres deform based on the excellent photo-thermal characteristics of the agglomerated gold nanospheres, a welding effect is generated between adjacent gold nanospheres, and a strong surface local plasma enhancement effect is generated around a welding point, so that the fluorescence of the gold nanospheres is greatly enhanced. The invention uses 405 nanometer continuous laser to irradiate the gold nanosphere with the diameter of 160 nanometers for 4.5 seconds continuously, so that the fluorescence of the gold nanosphere is enhanced by more than 150 times. The invention also realizes the continuous adjustment of the fluorescence spectrum of the gold nanospheres.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: how to provide a method and a device for rapidly and controllably realizing fluorescence enhancement of gold nanospheres through continuous laser, which can realize continuous adjustment of fluorescence spectra of the gold nanospheres.
The technical scheme adopted by the invention is as follows: a method for enhancing fluorescence intensity of gold nanospheres is characterized in that continuous laser is focused on agglomerated gold nanosphere particles (4) with the diameter of 150-170 nanometers to excite the agglomerated gold nanosphere particles (4) to obtain gold nanosphere fluorescence, the gold nanosphere fluorescence is enhanced along with the increase of laser irradiation time, the optimal enhancement effect is obtained when the laser irradiation time reaches 4.5 seconds, and then the gold nanosphere fluorescence is weakened along with the increase of the laser irradiation time.
A device for enhancing fluorescence intensity of gold nanospheres comprises a near ultraviolet continuous laser (1), a dichroic mirror (2), an objective lens (3), aggregated gold nanosphere particles (4), a cover glass (5), a beam splitter (6), a photodiode (7) and a spectrometer (8), wherein after passing through the objective lens (3), continuous laser emitted by the near ultraviolet continuous laser (1) is focused on the aggregated gold nanosphere particles (4) with the diameter of 150 plus 170 nanometers to excite the aggregated gold nanosphere particles (4) to obtain fluorescence of the gold nanospheres; the fluorescence of the gold nanospheres reversely passes through the objective lens (3) to form divergent fluorescence, the divergent fluorescence is detected by the photodiode (7) and the spectrometer (8), the fluorescence detected by the photodiode (7) and the spectrometer (8) is enhanced along with the increase of laser irradiation time, the fluorescence detected by the photodiode (7) and the spectrometer (8) begins to weaken after the fluorescence is increased to the optimal enhancement effect, and the fluorescence detected by the photodiode (7) and the spectrometer (8) obtains the maximum optimal enhancement effect when the laser irradiation time reaches 4.5 seconds.
As a preferred mode: the method for obtaining the agglomerated gold nanosphere particles (4) comprises the following steps of spin-coating a 150-170-nanometer gold nanosphere solution with the optical density of 0.005 on a cover glass with the thickness of 0.17 mm, wherein the spin-coating parameters are as follows: firstly spin-coating for 10 seconds at the rotating speed of 500 revolutions per minute, then spin-coating for 20 seconds at the rotating speed of 2000 revolutions per minute, and finally spin-coating for 10 seconds at the rotating speed of 500 revolutions per minute, wherein the distance between the gold nanospheres is 9-11 nanometers.
As a preferred mode: the power of the near ultraviolet continuous laser (1) is 2 milliwatts, the wavelength of the emitted continuous laser is 405 nanometers, the magnification of the objective lens (3) is 100 times, and the numerical aperture is 1.3.
The principle of the invention is as follows: the gold nanospheres have a local surface plasmon enhancement effect, the fluorescence intensity of the gold nanospheres can be enhanced, and the enhancement effect is influenced by the surface roughness of the gold nanospheres. The surface of the gold nanosphere chemically synthesized is relatively smooth, so that the enhancement effect is weak. When the gold nanospheres are irradiated by continuous laser, the gold nanospheres are rapidly heated due to the excellent photo-thermal conversion effect, so that the surfaces of the gold nanospheres are melted and deformed. When the adjacent gold nanospheres melt, the gold nanospheres contact with each other to form a sharp welding point, so that a strong local surface plasmon enhancement effect is generated, and the fluorescence intensity of the gold nanospheres is greatly enhanced.
The invention has the beneficial effects that: according to the invention, the agglomerated gold nanospheres are irradiated by near ultraviolet continuous laser, so that adjacent gold nanospheres generate a welding effect under the action of a photo-thermal effect, and further a strong local surface plasmon enhancement effect is generated, so that the fluorescence intensity of the gold nanospheres is enhanced by more than 150 times. Compared with the existing method for forming the gold nanosphere dimer, the method has the advantages of low cost, simple process, suitability for large-area industrial operation and the like. Compared with the method of adding other noble metal nano particles, the method has better fluorescence enhancement effect. The invention effectively solves the problem of low fluorescence efficiency of the gold nanospheres, so that the gold nanospheres with strong fluorescence emission capability can be applied to the aspects of biological imaging, medical diagnosis, treatment and the like, the imaging definition and the diagnosis accuracy are improved, and the treatment effect is enhanced. Compared with the fixed fluorescence enhancement effect obtained by adding gold nanosphere dimers or other noble metal nanoparticles, the enhancement effect of the invention on the fluorescence of the gold nanospheres changes along with the irradiation time, and the irradiation can be stopped when the required fluorescence intensity is obtained according to the actual requirement. Therefore, the method has higher flexibility for enhancing the fluorescence of the gold nanospheres, has wider application range and can be selected and used according to actual conditions. When the agglomerated gold nanospheres are irradiated by continuous laser to enhance the fluorescence intensity of the agglomerated gold nanospheres, the fluorescence spectrum of the gold nanospheres changes, and the degree of the change of the fluorescence spectrum depends on the time of continuous laser irradiation. Therefore, the fluorescence spectrum of the gold nanosphere with continuous adjustment can be obtained, and the method has important application in aspects such as color display and the like.
Drawings
FIG. 1 is a schematic structural diagram of a device for enhancing fluorescence intensity of gold nanospheres according to the present invention;
FIG. 2 is a transmission electron microscope characterization of gold nanosphere samples prepared by spin coating;
FIG. 3 is a graph showing the relationship between the fluorescence intensity of gold nanospheres under 405 nm continuous laser irradiation and the irradiation time;
FIG. 4 is a graph showing the relationship between the fluorescence enhancement factor of gold nanospheres under the irradiation of 405 nm continuous laser and the irradiation time;
FIG. 5 is a graph showing the relationship between the fluorescence spectrum of gold nanospheres and the change of the peak position with irradiation time under the irradiation of 405 nm continuous laser;
in the figure: the system comprises a 1-near ultraviolet continuous laser, a 2-dichroic mirror, a 3-objective lens, a 4-agglomerated gold nanosphere, a 5-cover glass, a 6-beam splitter, a 7-photodiode and an 8-spectrometer.
Detailed Description
As shown in fig. 1, the apparatus for enhancing fluorescence intensity of gold nanospheres in this embodiment includes a 1-laser, a 2-dichroic mirror, a 3-objective lens, 4-agglomerated gold nanospheres, a 5-cover glass, a 6-beam splitter, a 7-photodiode, and an 8-spectrometer. The laser 1 is a continuous laser with the wavelength of 405 nanometers, the dichroic mirror 2 is arranged on an emergent light path of the laser 1 and used for reflecting laser to the objective lens 3, and the objective lens 3 is positioned on a reflected light path of the dichroic mirror 2; the objective lens 3 collects fluorescence emitted by the agglomerated gold nanospheres 4 while focusing laser, the generated fluorescence is transmitted to the beam splitter 6 through the dichroic mirror 2, the beam splitter 6 is arranged on a transmission light path of the dichroic mirror 2, and the directions of a reflection light path and a transmission light path of the dichroic mirror 2 are opposite; fluorescence produced by the agglomerated gold nanospheres 4 is split by the beam splitter 6 and is detected by the photodiode 7 and the spectrometer 8 respectively, the photodiode 7 is arranged on a transmission light path of the beam splitter 6, the spectrometer 8 is arranged on a reflection light path of the beam splitter 6, and the transmission light path of the beam splitter 6 and the reflection light path form a right-angle relationship.
The operating power of the laser 1 is 2 milliwatts.
The dichroic mirror 2 can realize the reflection of 405 nm laser light and the transmission of fluorescence with the wavelength larger than 420 nm.
The magnification of the objective lens 3 is × 100, and the numerical aperture is NA = 1.3.
The beam splitting ratio of the beam splitter 6 to the fluorescence is 1:9, wherein the weaker fluorescence is positioned on a transmission light path, and the stronger fluorescence is positioned on a reflection light path.
The photodiode 7 is used for observing the fluorescence intensity of the gold nanometer in real time.
The spectrometer 8 is used for collecting the fluorescence spectrum of the gold nanospheres in real time.
Example 2
In this embodiment, a method for enhancing fluorescence intensity of gold nanospheres includes the following steps:
1) preparation of agglomerated gold nanosphere sample 4:
and (3) taking a cover glass, respectively cleaning the cover glass by using acetone, potassium hydroxide and deionized water, and spin-coating the gold nanosphere solution on the cover glass by using a spin-coating method to prepare an agglomerated gold nanosphere sample.
The thickness of the cover glass is 0.17 mm.
The gold nanospheres are prepared by a seed-mediated growth method, the diameter of the obtained gold nanospheres is about 160 nanometers, and the optical density of a gold nanosphere solution used for spin coating is 0.005.
The specific parameters of the spin-coating method are as follows: firstly, spin-coating for 10 seconds at a rotating speed of 500 revolutions per minute; secondly, spin-coating for 20 seconds at the rotating speed of 2000 rpm; thirdly, spin-coating for 10 seconds at a rotating speed of 500 rpm.
Fig. 2 shows a transmission electron microscope characterization result of the prepared aggregated gold nanosphere sample, which shows that a large number of gold nanospheres are gathered together in a visual field, the diameter of each gold nanosphere is about 160 nm, the distance between adjacent gold nanospheres is about 10 nm, the gold nanospheres are well dispersed on the surface of the cover glass, and the gold nanospheres are not stacked in the direction perpendicular to the cover glass.
2) Irradiating the agglomerated gold nanospheres 4 with a 405 nm continuous laser 1:
turning on the 405 nm continuous laser 1, and adjusting the laser power to 2 milliwatts; the laser emission direction is adjusted to pass through the objective lens. And opening the photodiode 7, adjusting the distance between the cover glass 5 and the objective lens 3, observing the signal intensity output by the photodiode 7, and when the signal intensity reaches the maximum, indicating that the laser is focused on the surface of the gold nanosphere on the cover glass.
3) Observing the fluorescence intensity of the gold nanospheres, and collecting the fluorescence spectrum of the gold nanospheres:
and (3) opening the photodiode 7 and the spectrometer 8, and observing the change of the fluorescence intensity and the fluorescence spectrum of the gold nanospheres under the condition that laser is focused on the surface of the gold nanospheres and continuous laser irradiation of 405 nanometers.
FIG. 3 shows the trace of the fluorescence intensity of gold nanospheres under the irradiation of 405 nm continuous laser light, which is detected by the photodiode, as a function of the irradiation time. As can be seen from the figure, the fluorescence intensity of the gold nanospheres is not obviously changed and is stabilized at 0.55 multiplied by 10 in the first 0.5 seconds of the 405 nanometer continuous laser irradiation4One (fluorescence photon)/second; then the fluorescence intensity of the gold nanospheres is obviously enhanced, and the fluorescence becomes stronger with the increase of the irradiation time; at 4.5 seconds, the fluorescence intensity reached a maximum of about 8.5X 105One/second; with the further irradiation of the 405 nanometer continuous laser, the fluorescence of the gold nanospheres shows a certain attenuation trend; after the 405 nanometer continuous laser is turned off for 5.35 seconds, the fluorescence of the gold nanospheres is also immediately turned off.
To further illustrate the fluorescence enhancement effect of gold nanospheres, the fluorescence enhancement factor is defined herein as: 405 nm continuous laser irradiationtAfter time, the ratio of its fluorescence intensity to the initial fluorescence intensity. FIG. 4 shows the fluorescence enhancement factor of gold nanospheres as a function of 405 nm continuous laser irradiation time. As can be seen from the figure, the fluorescence intensity of the gold nanospheres showed a rapid increase between 1 second and 4 seconds after the 405 nm continuous laser irradiation: after laser irradiation for 2 seconds, the fluorescence intensity of the gold nanospheres increased by 30 timesThe above step (1); after the irradiation for 3 seconds, the fluorescence intensity is increased by more than 90 times; after 4 seconds of irradiation, the fluorescence intensity increased nearly 150-fold. The fluorescence intensity was relatively stable during the laser irradiation for 4 to 5 seconds, and then exhibited some attenuation.
Further, as the fluorescence intensity of the gold nanospheres is enhanced, the fluorescence spectrum of the gold nanospheres is obviously changed. FIG. 5a shows fluorescence spectra of gold nanospheres after 405 nm continuous laser irradiation for different time periods; it was found that the fluorescence peak (where the fluorescence spectrum is most intense) shifts to longer wavelengths as the illumination time increases. FIG. 5b shows the change of the fluorescence peak with irradiation time; it was found that the illumination time was between 0 and 3 seconds, and the fluorescence peak gradually increased from 645 nm to 710 nm; the irradiation time is 3 seconds to 5 seconds, and the position of the fluorescence peak is basically unchanged.
Claims (1)
1. A method for enhancing fluorescence intensity of gold nanospheres is characterized by comprising the following steps: focusing and exciting the agglomerated gold nanospheres (4) on the agglomerated gold nanospheres (4) by using continuous laser with the power of 2 milliwatts and the wavelength of 405 nanometers to obtain the fluorescence of the agglomerated gold nanospheres (4), wherein when the agglomerated gold nanospheres (4) are irradiated by the continuous laser, because the gold nanosphere particles (4) have excellent photo-thermal conversion effect, the temperature of the gold nanosphere particles is rapidly increased, so that the surfaces of the gold nanosphere particles are melted, further generates deformation, when the adjacent gold nanosphere particles (4) are melted, the gold nanosphere particles are contacted with each other to form a sharp welding point, further generating strong local surface plasma enhancement effect, greatly enhancing the fluorescence intensity of the agglomerated gold nanosphere particles (4), the fluorescence of the agglomerated gold nanosphere particles (4) is enhanced along with the increase of the laser irradiation time, the best enhancement effect is obtained when the laser irradiation time reaches 4.5 seconds, and then is weakened along with the increase of the laser irradiation time; the method for obtaining the agglomerated gold nanosphere particles (4) comprises the following steps of spin-coating a 150-170-nanometer gold nanosphere solution with the optical density of 0.005 on a cover glass with the thickness of 0.17 mm, wherein the spin-coating parameters are as follows: firstly spin-coating for 10 seconds at the rotating speed of 500 revolutions per minute, then spin-coating for 20 seconds at the rotating speed of 2000 revolutions per minute, and finally spin-coating for 10 seconds at the rotating speed of 500 revolutions per minute, wherein the distance between the gold nanospheres is 9-11 nanometers.
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