Fluorescent molecule for rapidly measuring spot size and light intensity distribution of exciting light and preparation method of fluorescent molecule

Abstract

The invention discloses a method for simply and quickly measuring the size of a light spot, which adopts the principle that high-fluorescence quantum efficiency molecules with relatively poor light stability shown in a formula I are coated on a two-dimensional substrate, and a fluorescent molecular film with high luminescence and easy photobleaching is irradiated by a light spot to be measured, so that the size of the light spot is measured. And secondary imaging is carried out under a fluorescence microscope by utilizing the fluorescence quenching phenomenon of the irradiated fluorescent molecules, so that rapid measurement of the spot size and the light intensity distribution is realized. # imgabs0 #

Description

Technical Field

[0001] The invention relates to the field of organic optoelectronics and optical measurement technology, and in particular to the development of highly luminescent and easily photobleached fluorescent molecules, which can be used for rapid measurement of excitation light spot size and light intensity distribution. Technical Background

[0002] The spot size is one of the important factors that determine the light power density, optical imaging quality and laser processing accuracy. Accurately measuring the spot size of different light sources can help optimize the fluorescence imaging results and improve the laser processing resolution. At present, the commonly used methods for measuring the spot size include diffraction pattern analysis, knife-edge method and beam camera direct imaging method. The diffraction pattern analysis method is a method commonly used to measure the spot size in laser systems. Its principle is that the light beam is guided through a small hole or a small slit and its diffraction pattern is projected on the detector. By analyzing and processing the diffraction pattern on the detector, the spot size can be calculated. The test accuracy of the beam quality analyzer developed based on this method can reach the micron level, but the measurement range of the spot size can only reach a few hundred microns at most. In addition, the price of the beam quality analyzer is relatively expensive and the cost of use is high. The knife-edge analysis method measures the spot size by scanning the light beam with a knife edge while measuring the light power of the light passing through with a photodiode. First, measure the total beam power, then gradually cut the beam with a knife edge to measure the position corresponding to the light intensity of different intensities. Then, twice the distance between the two positions is equal to 1 / e 2 The diameter of the spot can be calculated from the diameter. Although the knife-edge analysis method has a relatively simple device and algorithm, it can only measure the diameter of a circular Gaussian beam and cannot obtain the laser intensity profile information. The camera direct imaging method requires the use of a beam camera, which can measure the size and intensity profile of any beam in principle. However, since the high-sensitivity sensor chip needs to prevent the imaging intensity from being oversaturated and damaging the instrument, and background light compensation is required during measurement, the subsequent spot size calculation requires a large amount of data processing, which greatly increases the difficulty and cost of measuring the spot size. Summary of the invention

[0003] One of the purposes of the present invention is to provide a class of fluorescent molecules for rapid measurement of excitation light spot size and light intensity distribution.

[0004] The second object of the present invention is to provide a method for preparing a type of fluorescent molecules and a uniform luminescent film for rapidly measuring the spot size and light intensity distribution of an excitation light.

[0005] A third object of the present invention is to provide a method for rapidly measuring the spot size and light intensity distribution of an excitation light.

[0006] The present invention first provides a compound shown in the following formula I:

[0007]

[0008] Wherein, R is the same or different and is independently selected from H, C 1-12 Alkyl, C 1-12 Alkoxy, the C 1-12 Alkyl and C 1-12 The alkoxy group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-12 Alkyl, C 1-12 Alkoxy;

[0009] R1 is a non-hydrogen substituent selected from halogen, OH, C 1-12 Alkoxy or C 1-12 Alkyl, the C 1-12 Alkoxy or C 1-12 The alkyl group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-12 Alkoxy; the number of the non-hydrogen substituents R1 is 0, 1, 2, 3 or 4;

[0010] R2 is selected from -C 6-12 Aryl-5-12 membered heteroaryl, -5-12 membered heteroaryl-5-12 membered heteroaryl, -C 6-12 Aryl-C 6-12 Aryl or -5-12 membered heteroaryl-C 6-12 Aryl; said C 6-12 Aryl and 5-12 membered heteroaryl are optionally substituted by one, two or more of the following groups: halogen, OH, C 1-12 Alkyl, C 1-12 Alkoxy;

[0011] (x+z) is the number of fluorenyl or substituted fluorenyl groups, which is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0012] y is the number of benzene rings or substituted benzene rings, which is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0013] According to an embodiment of the present invention, R is the same or different and is independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy; the C 1-6 Alkyl and C 1-6 The alkoxy group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy;

[0014] R1 is a non-hydrogen substituent selected from halogen, OH, C 1-6 Alkoxy or C 1-6 Alkyl, the C 1-6 Alkoxy or C1-6 The alkyl group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-6 Alkoxy; the number of the non-hydrogen substituents R1 is 0, 1 or 2;

[0015] R2 is selected from -phenyl-5-12 membered heteroaryl, -5-12 membered heteroaryl-5-12 membered heteroaryl, -phenyl-phenyl or -5-12 membered heteroaryl-phenyl; the phenyl and 5-12 membered heteroaryl are optionally substituted by one, two or more of the following groups: halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy;

[0016] (x+z) is the number of fluorenyl or substituted fluorenyl groups, which is 1, 2, 3, 4 or 5;

[0017] y is the number of benzene rings or substituted benzene rings, which is 0, 1, 2 or 3.

[0018] According to an embodiment of the present invention, R is the same or different and is independently selected from H or C 1-6 alkyl.

[0019] According to an embodiment of the present invention, (x+z) is the number of fluorenyl groups or substituted fluorenyl groups, which is 1, 2 or 3;

[0020] According to an embodiment of the present invention, y is the number of benzene rings or substituted benzene rings, which is 0 or 1.

[0021] In some embodiments, the compound represented by Formula I is selected from the following structures represented by Formula IA or IB:

[0022]

[0023] Wherein R is the same or different and is independently selected from H or C 1-6 alkyl;

[0024] R2 are the same or different and are independently selected from -phenyl-imidazole, -phenyl-benzimidazole, -phenyl-indole, -phenyl-oxazole, -phenyl-benzoxazole, -phenyl-pyrrole, -phenyl-indene or -phenyl-benzothiophene;

[0025] x is 1, 2, or 3.

[0026] In some specific embodiments, one R is H and the other is hexyl; or both R are hexyl; or both R are H.

[0027] In some specific embodiments, x is 3.

[0028] In some specific embodiments, R2 is the same and is selected from a substituent consisting of a benzene ring and the following groups:

[0029]

[0030] Among them, the * is the site connected to the phenyl group.

[0031] In some specific embodiments, R2 is the same and is selected from -phenyl-benzimidazole.

[0032] In some specific embodiments of the present invention, the compound represented by formula I is selected from the following structures:

[0033]

[0034] According to an embodiment of the present invention, the compound shown in Formula I is a highly luminescent, easily photobleached fluorescent molecule, and its absorption range is 300-410 nanometers. When it is used to measure the excitation light spot size and / or light intensity distribution, the wavelength range of the light source to be measured should include the absorption range of the compound shown in Formula I, so that the fluorescent molecule is photobleached under the irradiation of the light source to be measured.

[0035] According to an embodiment of the present invention, the fluorescence quantum efficiency of the compound shown in Formula I is greater than 90%. The compound has high fluorescence emission efficiency and can increase the contrast between the photobleached area and the non-photobleached area during fluorescence imaging.

[0036] According to an embodiment of the present invention, the emission range of the compound represented by Formula I is 400-480 nanometers. The emission wavelength of the compound is within the visible light range, which enables direct observation of fluorescence imaging under a fluorescence microscope.

[0037] The present invention also provides a method for preparing the compound represented by formula I, comprising the following steps:

[0038]

[0039] 1) Compound IA reacts with compound IB to obtain compound IC;

[0040] 2) Compound IA reacts with compound ID to obtain compound IE;

[0041] 3) Compound IC, IE react with compound IF to obtain compound I;

[0042] Wherein, R, R1, R2, x, y, z have the above definitions;

[0043] L is a halogen.

[0044] The present invention also provides the use of the compound shown in the above formula I in preparing fluorescent molecular films.

[0045] The present invention also provides a method for preparing a fluorescent molecular film using the compound shown in the above formula I, comprising:

[0046] Dissolving the compound represented by formula I in an organic solvent, dropping the obtained solution on a two-dimensional substrate, and drying to obtain a fluorescent film;

[0047] The organic solvent is alcohol or chloroform, or a mixed solvent of alcohol and chloroform.

[0048] According to an embodiment of the present invention, the concentration of the compound represented by Formula I in the solution is 0.001 to 10.0 mg / ml, and the sampling volume is 0.01 to 10.0 μl.

[0049] According to an embodiment of the present invention, the alcohol is any one or a mixed solvent of volatile alcohols such as ethanol, n-propanol, isopropanol, n-butanol, isobutanol, n-pentanol, isopentane and n-hexanol.

[0050] According to an embodiment of the present invention, the two-dimensional substrate may be one of a glass sheet, a quartz sheet, a silicon sheet, a mica sheet or graphite.

[0051] According to an embodiment of the present invention, in the method for preparing the fluorescent film, the key to obtaining a dense and uniform fluorescent film with high luminescence and easy photobleaching is to quickly remove the solvent in the uniformly dispersed droplets.

[0052] The present invention also provides a method for using the fluorescent film as described above to measure the spot size of excitation light, and the measurement method includes the following steps: irradiating the fluorescent molecular film as described above with the light source to be measured, performing fluorescence imaging using the fluorescence quenching phenomenon of the fluorescent molecules, and then performing grayscale analysis on the fluorescence imaging image to obtain a two-dimensional distribution of grayscale values, and obtaining the spot size of the light source to be measured based on the distribution of grayscale values.

[0053] The present invention also provides a method for using the fluorescent film as described above to measure the intensity distribution of excitation light, and the measurement method includes the following steps: irradiating the fluorescent molecular film as described above with the light source to be measured, performing fluorescence imaging using the fluorescence quenching phenomenon of the fluorescent molecules, performing grayscale analysis on the fluorescence imaging image after photobleaching to obtain axial grayscale changes, and obtaining the intensity distribution of the light source to be measured based on the axial changes in grayscale.

[0054] According to the embodiment of the present invention, the optical parameter conditions of the light source to be measured are: the wavelength range includes 300-410 nanometers, so that the fluorescent molecule undergoes photobleaching under the irradiation of the light source to be measured; the light power range is 10 -3 -10 3 mW, single point irradiation time 10 -2 -10 2 Second.

[0055] According to an embodiment of the present invention, when the wavelength of the light source to be measured meets the requirements, the smaller the optical power density, the longer the time required for photobleaching. The optical power density threshold of the light source to be measured is required to be 10 milliwatts per square millimeter to achieve rapid measurement of the spot size and / or intensity distribution of the excitation light.

[0056] According to an embodiment of the present invention, the light source to be tested can achieve photobleaching of fluorescent molecules with high luminescence and easy photobleaching above the energy intensity threshold, and can be any one of LED light source, halogen lamp, laser, mercury lamp, xenon lamp, etc.

[0057] The method for rapidly measuring the spot size and / or intensity distribution of the present invention irradiates a highly luminescent, easily photobleached fluorescent molecular film with a light source to be measured (LED light source, halogen lamp, laser, mercury lamp, xenon lamp, etc.), and uses the fluorescence quenching phenomenon of the irradiated highly luminescent, easily photobleached fluorescent molecules to perform fluorescence imaging, thereby realizing rapid measurement of the spot size and / or intensity distribution of the light source to be measured.

[0058] The above method for quickly measuring the spot size and intensity distribution of a light source can be specifically as follows: when the light source to be measured (LED light source, halogen lamp, laser, mercury lamp, xenon lamp, etc.) is irradiated on a highly luminescent, easily photobleached fluorescent molecular film, the highly luminescent, easily photobleached fluorescent molecules will undergo fluorescence quenching under the irradiation of the light source to be measured, forming an obvious dark quenching area; then, fluorescence imaging of the highly luminescent, easily photobleached fluorescent molecular film is performed under the microscope fluorescence mode and grayscale analysis is performed to obtain the spot size and intensity distribution of the light source to be measured.

[0059] Beneficial Effects

[0060] Compared with the existing methods for measuring spot size and intensity distribution, the present invention provides a simple and rapid method for measuring spot size and light intensity distribution. The principle is to coat a high fluorescence quantum efficiency molecule with relatively poor photostability as shown in Formula I on a two-dimensional substrate, irradiate the highly luminescent and easily photobleached fluorescent molecule film with a light spot to be measured, and perform secondary imaging under a fluorescence microscope by utilizing the fluorescence quenching phenomenon of the irradiated fluorescent molecules, thereby realizing rapid measurement of the spot size and light intensity distribution.

[0061] The method of the invention is simple and quick to operate and has certain application prospects.

[0062] Definitions and explanations of terms

[0063] In this document, the term "halogen" means fluorine, chlorine, bromine and / or iodine. Correspondingly, the term "halo" refers to fluoro, chloro, bromo and / or iodo. Within the scope of this document, when an atom, a radical or a part is halogenated, the atom at the halogenated position may be monosubstituted, disubstituted or polysubstituted up to full substitution by halogen atoms, for example, "halo C 1-12 alkyl".

[0064] The term "C 1-12 "Alkyl" is understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 12 carbon atoms, preferably C 1-6 Alkyl. "C 1-6 “Alkyl” is understood as meaning preferably a linear or branched, saturated, monovalent hydrocarbon radical having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl radical is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers. In particular, the radical has 1, 2 or 3 carbon atoms (“C 1-3 alkyl"), for example methyl, ethyl, n-propyl or isopropyl.

[0065] The term "C 3-12 "Cycloalkyl" should be understood to mean a saturated monovalent monocyclic, bicyclic hydrocarbon ring or polycyclic hydrocarbon ring (also called condensed hydrocarbon ring) having 3-12 carbon atoms. Bicyclic or polycyclic cycloalkyl groups include paracyclic cycloalkyl, bridged cycloalkyl and spirocyclic cycloalkyl; the paracyclic refers to a condensed ring structure formed by two or more cyclic structures sharing two adjacent ring atoms (i.e., sharing one bond). The bridged ring refers to a condensed ring structure formed by two or more cyclic structures sharing two non-adjacent ring atoms. The spirocyclic refers to a condensed ring structure formed by two or more cyclic structures sharing one ring atom. For example, the C 3-20 The cycloalkyl group may be C 3-8 Monocyclic cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, or C 7-12 Cycloalkyl ring, such as decalin ring.

[0066] The term "3-12 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring containing 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclyl may be connected to the rest of the molecule through any one of the carbon atoms or the nitrogen atom (if present). In particular, the heterocyclyl may include, but is not limited to, a 4-membered ring such as azetidinyl, oxetanyl, a 5-membered ring such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl, or a 7-membered ring such as diazepanyl. Optionally, the heterocyclyl may be benzo-fused. The heterocyclic group may be bicyclic, for example, but not limited to, a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrol-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The ring containing the nitrogen atom may be partially unsaturated, i.e., it may contain one or more double bonds, for example, but not limited to, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 4,5-dihydrooxazolyl, or 4H-[1,4]thiazinyl, or it may be benzo-fused, for example, but not limited to, dihydroisoquinolinyl.

[0067] The term "C 6-12 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 or a ring having 9 carbon atoms ("C9 aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 aryl), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 aryl), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 "aryl"), for example anthracenyl.

[0068] The term "5-12 membered heteroaryl" is to be understood as including monovalent mono-, bi- or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which contain 1 to 5, preferably 1 to 3 heteroatoms which are each independently selected from N, O and S and which, in addition, in each case may be benzo-fused. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and the like and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl and the like; or azinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl and the like.

[0069] Unless otherwise indicated, heterocyclyl, heteroaryl or heteroarylene includes all possible isomeric forms thereof, such as positional isomers thereof. Thus, for some illustrative non-limiting examples, pyridyl or pyridylene includes pyridine-2-yl, pyridine-2-yl, pyridine-3-yl, pyridine-3-yl, pyridine-4-yl and pyridine-4-yl; thienyl or thienylene includes thien-2-yl, thien-2-yl, thien-3-yl and thien-3-yl.

[0070] The above term “C 1-12 The definition of "alkyl" also applies to compounds containing "C 1-12 Other terms for "alkyl", such as the term "halo-C 1-12 Alkyl" or "C 1-12 Alkoxy" and so on. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Figure 1 a) One-dimensional nuclear magnetic hydrogen spectrum of the highly luminescent and easily photobleached fluorescent molecule I-1 of Example 1 of the present invention. b) Mass spectrometry data spectrum, the relative molecular mass is 1127.321 g / mol. c) In a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) UV-visible absorption spectrum. d) in a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) in the fluorescence emission spectrum.

[0072] Figure 2a) One-dimensional nuclear magnetic hydrogen spectrum of the highly luminescent and easily photobleached fluorescent molecule I-2 of Example 2 of the present invention. b) Mass spectrometry data spectrum, the relative molecular mass is 1383.891 g / mol. c) In a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) UV-visible absorption spectrum. d) in a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) in the fluorescence emission spectrum.

[0073] Figure 3 a) Fluorescence optical microscope image of the highly luminescent and easily photobleached fluorescent molecule I-1 film of Example 3 of the present invention and b) grayscale distribution analysis. c) Fluorescence optical microscope image of the highly luminescent and easily photobleached fluorescent molecule I-1 film after irradiation with the light source to be tested of Example 4 of the present invention and d) grayscale distribution analysis.

[0074] Figure 4 a) Fluorescence optical microscope image of the highly luminescent and easily photobleached fluorescent molecule I-2 film of Example 5 of the present invention and b) grayscale distribution analysis. c) Fluorescence optical microscope image of the highly luminescent and easily photobleached fluorescent molecule I-2 film after irradiation with the light source to be tested of Example 5 of the present invention and d) grayscale distribution analysis. DETAILED DESCRIPTION

[0075] The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[0076] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0077] Example 1

[0078] Preparation and synthesis of highly luminescent and easily photobleached fluorescent molecule compound I-1

[0079] The synthetic route is as follows:

[0080]

[0081] 1) Synthesis steps of molecule 1: Weigh 2.00 g (1 equivalent) of 2-(4-bromophenyl)benzimidazole, 2.79 g (1.5 equivalents) of bis-pinacolato diboron, 2.87 g (4 equivalents) of potassium acetate and 0.54 g (0.1 equivalent) of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, add to 30 ml of 1,4-dioxane solution, introduce argon gas for 30 minutes, react at 80°C for 8 hours, remove the solvent under vacuum, extract and wash the organic phase with 3×40 ml of ethyl acetate and 80 ml of saturated brine, respectively, add anhydrous sodium sulfate for drying, and purify by column chromatography to obtain 2.01 g of product molecule 1.

[0082] 2) Synthesis steps of molecule 2: Take 2.00 g (1 equivalent) of the product molecule 1 obtained in step 1), 4.61 g (1.5 equivalents) of 2,7-dibromo-9,9-dihexylfluorene and 0.72 g (0.1 equivalents) of tetrakis(triphenylphosphine)palladium, add them to 30 ml of 1,4-dioxane solution, add 6 ml of 2.0 mmol / L potassium carbonate aqueous solution under stirring, pass argon for 30 minutes, heat to 80°C, and stir for 8 hours. After removing the solvent under vacuum, extract and wash the organic phase with 3×40 ml of chloroform and 80 ml of saturated brine, respectively, add anhydrous sodium sulfate to dry, and purify by column chromatography to obtain 3.08 g of product molecule 2.

[0083] 3) Synthesis steps of molecule 3: 1.00 g (1 equivalent) of 1,4-dibromobenzene, 2.08 g (5 equivalents) of potassium acetate, 3.23 g (3 equivalents) of bis-pinacolyl diboron and 0.31 g (0.1 equivalent) of [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium were added to 40 ml of 1,4-dioxane solution, argon was introduced for 30 minutes, heated to 80°C, and stirred for 8 hours. After removing the solvent under vacuum, the organic phase was extracted and washed with 3×45 ml of ethyl acetate and 100 ml of saturated brine, respectively, and dried by adding anhydrous sodium sulfate. Purification by column chromatography gave 1.01 g of product molecule 3.

[0084] 4) Synthesis steps of molecule I-1: Take 1.00 g (2.5 equivalents) of the product molecule 2 obtained in step 2), 0.22 g (1 equivalent) of the product molecule 3 obtained in step 3) and 0.08 g (0.1 equivalent) of tetrakis(triphenylphosphine)palladium, add to 30 ml of 1,4-dioxane solution, add 6 ml of 2.0 mmol / L potassium carbonate aqueous solution under stirring, pass argon for 30 minutes, heat to 80°C, and stir for 8 hours. After removing the solvent under vacuum, extract and rinse the organic phase with 3×35 ml of chloroform and 40 ml of saturated saline, respectively, add anhydrous sodium sulfate to dry, and purify by column chromatography to obtain 0.61 g of the target high-luminescence, easy-to-photobleach fluorescent molecule I-1. Its nuclear magnetic resonance data is shown in the figure below Figure 1a; mass spectrum data as shown in Figure 1 b; in a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) in the UV-visible absorption spectrum as shown in Figure 1 c; the fluorescence emission spectrum is shown in Figure 1 As shown in d.

[0085] Example 2

[0086] Preparation and synthesis of highly luminescent and easily photobleached fluorescent molecule compound I-2

[0087] The synthetic route is as follows:

[0088]

[0089] 5) Synthesis steps of molecule 5: 1.00 g (1 equivalent) of 2,7-dibromo-9,9-dihexylfluorene, 1.00 g (5 equivalents) of potassium acetate, 1.55 g (3 equivalents) of bis-pinacolyl diboron and 0.15 g (0.1 equivalent) of [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium were added to 40 ml of 1,4-dioxane solution, argon was introduced for 30 minutes, heated to 80°C, and stirred for 8 hours. After removing the solvent under vacuum, the organic phase was extracted and washed with 3×45 ml of ethyl acetate and 100 ml of saturated brine, respectively, and dried by adding anhydrous sodium sulfate. Purification by column chromatography gave 0.96 g of product molecule 5.

[0090] 6) Synthesis steps of molecule I-2: Take 1.00 g (2.5 equivalents) of the product molecule 2 obtained in step 2), 0.39 g (1 equivalent) of the product molecule 5 obtained in step 5) and 0.08 g (0.1 equivalent) of tetrakis(triphenylphosphine)palladium, add to 30 ml of 1,4-dioxane solution, add 6 ml of 2.0 mmol / L potassium carbonate aqueous solution under stirring, pass argon for 30 minutes, heat to 80°C, and stir for 8 hours. After removing the solvent under vacuum, extract and rinse the organic phase with 3×35 ml of chloroform and 40 ml of saturated saline, respectively, add anhydrous sodium sulfate to dry, and purify by column chromatography to obtain 0.68 g of the target high-luminescence, easy-to-photobleach fluorescent molecule compound I-2. Its nuclear magnetic resonance data is shown in the figure below Figure 2 a; mass spectrum data as shown in Figure 2 b; in a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) in the UV-visible absorption spectrum as shown in Figure 2 c; the fluorescence emission spectrum is shown in Figure 2 As shown in d.

[0091] Example 3

[0092] Preparation of highly luminescent and easily photobleached fluorescent molecular films

[0093] 1) 0.1 mg of the highly luminescent and easily photobleached fluorescent molecule compound I-1 prepared in Example 1 was completely dissolved in 5 ml of a mixed solvent of butanol and chloroform (V 丁醇：氯仿 =1:1) to obtain a solution with a mass concentration of 0.02 mg / ml;

[0094] 2) Take 2 μl of the above solution and drop it on a glass slide, and dry it on a hot plate at 65° C. to obtain a dense and uniform fluorescent molecular film that is highly luminescent and easily photobleached.

[0095] Fluorescence optical microscope image and grayscale distribution analysis of the high fluorescence luminescent molecular film before irradiation by the light source to be tested Figure 3 a and 3b.

[0096] Example 4

[0097] The high-luminescence, easily photobleached fluorescent molecular film of Example 3 was irradiated with the LED light source to be tested at 100% intensity for 20 seconds. The wavelength was 365 nanometers. The light source was emitted through the 40x objective lens of the microscope. The actual measured light power was 10.27 milliwatts. Photobleaching occurred at the irradiation point of the light source to form dark spots.

[0098] Under the microscope fluorescence mode (mercury lamp light source), the fluorescent molecular film with high luminescence and easy photobleaching after photobleaching is imaged and grayscale analyzed to obtain the spot size and intensity distribution of the light source to be tested. Figure 3 c and 3d. After analysis and calculation, the measured excitation light diameter is about 533 microns, and the light power density is about 46 milliwatts per square millimeter.

[0099] Example 5

[0100] The fluorescent molecular film of compound I-2 was prepared by referring to the method of Example 3 above. The fluorescence optical microscope image and grayscale distribution analysis of the highly fluorescent luminescent molecule I-2 film before irradiation by the test light source are as follows: Figure 4 a and 4b.

[0101] The same light source is used to illuminate the fluorescent molecular film in Reference Example 4. The fluorescence optical microscope image and grayscale distribution analysis of the fluorescent molecule I-2 film with high luminescence and easy photobleaching after irradiation by the test light source are as follows: Figure 4 c and 4d. After analysis and calculation, the measured excitation light diameter is about 548 microns, and the light power density is about 44 milliwatts per square millimeter.

[0102] The above is an explanation of the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. The compound shown in formula I: in, R are the same or different and are independently selected from H, C 1-12 Alkyl, C 1-12 Alkoxy, the C 1-12 Alkyl and C 1-12 The alkoxy group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-12 Alkyl, C 1-12 Alkoxy; R1 is a non-hydrogen substituent selected from halogen, OH, C 1-12 Alkoxy or C 1-12 Alkyl, the C 1-12 Alkoxy or C 1-12 The alkyl group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-12 Alkoxy; the number of the non-hydrogen substituents R1 is 0, 1, 2, 3 or 4; R2 is selected from -C 6-12 Aryl-5-12 membered heteroaryl, -5-12 membered heteroaryl-5-12 membered heteroaryl, -C 6-12 Aryl-C 6-12 Aryl or -5-12 membered heteroaryl-C 6-12 Aryl; said C 6-12 Aryl and 5-12 membered heteroaryl are optionally substituted by one, two or more of the following groups: halogen, OH, C 1-12 Alkyl, C 1-12 Alkoxy; (x+z) is the number of fluorenyl or substituted fluorenyl groups, which is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; y is the number of benzene rings or substituted benzene rings, which is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

2. The compound according to claim 1, characterized in that R are the same or different and are independently selected from H, C 1-6 Alkyl, C 1-6 Alkoxy; the C 1-6 Alkyl and C 1-6 The alkoxy group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy; R1 is a non-hydrogen substituent selected from halogen, OH, C 1-6 Alkoxy or C 1-6 Alkyl, the C 1-6 Alkoxy or C 1-6 The alkyl group is optionally substituted by one, two or more of the following groups: halogen, OH, C 1-6 Alkoxy; the number of the non-hydrogen substituents R1 is 0, 1 or 2; R2 is selected from -phenyl-5-12 membered heteroaryl, -5-12 membered heteroaryl-5-12 membered heteroaryl, -phenyl-phenyl or -5-12 membered heteroaryl-phenyl; the phenyl and 5-12 membered heteroaryl are optionally substituted by one, two or more of the following groups: halogen, OH, C 1-6 Alkyl, C 1-6 Alkoxy; (x+z) is the number of fluorenyl or substituted fluorenyl groups, which is 1, 2, 3, 4 or 5; y is the number of benzene rings or substituted benzene rings, which is 0, 1, 2 or 3.

3. The compound according to claim 2, characterized in that The compound represented by formula I is selected from the structure represented by formula IA or IB: Wherein R is the same or different and is independently selected from H or C 1-6 alkyl; R2 are the same or different and are independently selected from -phenyl-imidazole, -phenyl-benzimidazole, -phenyl-indole, -phenyl-oxazole, -phenyl-benzoxazole, -phenyl-pyrrole, -phenyl-indene or -phenyl-benzothiophene; x is 1, 2, or 3.

4. The compound according to any one of claims 1 to 3, characterized in that The compound represented by formula I is selected from the following structures:

5. A method for preparing the compound according to any one of claims 1 to 4, characterized in that: The steps include: 1) Compound IA reacts with compound IB to obtain compound IC; 2) Compound IA reacts with compound ID to obtain compound IE; 3) Compound IC, IE react with compound IF to obtain compound I; Wherein, R, R1, R2, x, y, and z have the definitions as described in any one of claims 1 to 4; L is a halogen.

6. Use of the compound according to any one of claims 1 to 4 in the preparation of fluorescent molecular films.

7. A method for preparing a fluorescent molecular film using the compound according to any one of claims 1 to 4, characterized in that: include: Dissolving the compound represented by formula I in an organic solvent, dropping the obtained solution on a two-dimensional substrate, and drying to obtain a fluorescent film; The organic solvent is alcohol or chloroform, or a mixed solvent of alcohol and chloroform.

8. The method for measuring the spot size of excitation light using the fluorescent film as claimed in claim 7, characterized in that: The following steps are involved: The light source to be tested illuminates the fluorescent molecular film, and fluorescence imaging is performed using the fluorescence quenching phenomenon of the fluorescent molecules. Grayscale analysis is then performed on the fluorescence imaging image to obtain a two-dimensional distribution of grayscale values, and the spot size of the light source to be tested is obtained based on the distribution of grayscale values.

9. The method for measuring the intensity distribution of excitation light using the fluorescent film as claimed in claim 7, characterized in that: The following steps are involved: The light source to be tested is irradiated onto the fluorescent molecular film, fluorescence imaging is performed using the fluorescence quenching phenomenon of the fluorescent molecules, grayscale analysis is performed on the fluorescence imaging picture after photobleaching to obtain axial grayscale changes, and the intensity distribution of the light source to be tested is obtained according to the axial grayscale changes.

10. The method according to claim 8 or 9, characterized in that: The light source to be tested is any one of an LED light source, a halogen lamp, a laser, a mercury lamp, and a xenon lamp.

Images