A temperature measurement composition comprising biodegradable periodic mesoporous organic silica and at least two dyes.

Abstract

This specification provides a temperature-measuring composition comprising a plurality of biodegradable periodic mesoporous organosilica, B-PMO, and particles, wherein - each B-PMO particle is loaded with a first dye and a second dye, - both the first and second dyes are fluorescent, - fluorescence emission from the first dye triggers emission excitation of the second dye, and - the first and / or second dyes exhibit temperature-dependent fluorescence. Furthermore, methods for preparing such a temperature-measuring composition and methods for obtaining temperature data of cells, tissues, or objects using such a temperature-measuring composition are provided.

Description

[Technical Field]

[0001] Field of Invention The present invention is in the field of thermal monitoring and thermal mapping, and therefore more specifically, in the field of temperature measuring compositions that can be used. These temperature measuring compositions are preferably suitable for use as (nano)thermometers that can be used in vivo, preferably subcutaneously. [Background technology]

[0002] Background of the Invention Temperature is a fundamental quantity associated with many natural biological processes. Consequently, thermal monitoring plays a crucial role in disease diagnosis. While conventional techniques for temperature readout (e.g., thermocouples, thermistors, IR cameras) are reliable in many applications, they are either invasive or unable to detect temperatures below the skin surface, thus failing to adequately perform advanced in vivo and in vitro temperature mapping. The use of luminescence for in vivo temperature assessment has garnered considerable interest over the past decade. Since some materials possess luminescence properties that are strongly dependent on temperature, it is possible to develop luminescent (nano)thermometers. This allows for the design of sensors capable of nanoscale readout that can operate non-contact and minimally invasively. Such luminescent nanothermometers are promising novel tools for the early diagnosis and treatment control of inflammatory conditions or cancer (such cells have slightly higher temperatures) (e.g., during photothermal ablation, where the heating process must be carefully controlled to avoid damage to nearby healthy tissue). [Overview of the project] [Problems that the invention aims to solve]

[0003] However, considering the ultimate goal of clinical applications, many developed nanomaterials have potential toxicity and bioaccumulation in the human body, and therefore, despite their great potential and impact, luminescent nanothermometers have not yet progressed beyond the preclinical stage. Since most reported nanothermometers are currently based primarily on the incorporation of heavy metal ions rather than degradable materials, these aspects remain a real concern in the fields of nanomedicine, nanobiotechnology, nanotoxicology, and nanopharmacology.

[0004] Therefore, there is a need for temperature measuring compositions that are less toxic and preferably do not accumulate in the body, preferably suitable for use as nanothermometers. However, these temperature measuring compositions need to be highly sensitive and exhibit a high signal-to-noise ratio. Preferably, these temperature measuring compositions are easy to manufacture and allow for large variations in the wavelength, detection instrument, and excitation mode used. There is a particular need for temperature measuring compositions that can be used through the skin or through tissue. [Means for solving the problem]

[0005] Summary of the Invention The temperature measurement compositions and methods disclosed herein can satisfy one or more of the above requirements. The inventors have found that loading two fluorescent dyes (the emission of one dye exciting the other) into biodegradable PMO particles can satisfy at least some of the requirements formulated herein. Having two dyes in B-PMO particles ensures efficient energy transfer between the two dyes, making the entire system highly sensitive.

[0006] More specifically, the present invention provides a temperature measurement composition comprising a plurality of biodegradable periodic mesoporous organic silicas, B-PMO, and particles, wherein -Each B-PMO particle is loaded with the first dye and the second dye. -Both the first and second dyes are fluorescent. -Fluorescence emission from the first dye triggers emission excitation of the second dye, - The first dye and / or the second dye exhibit temperature-dependent fluorescence.

[0007] In some embodiments, both the first and second dyes are organic. In some embodiments, each B-PMO particle comprises multiple polymer chains of alkoxysilane of formula I.

[0008] -(RO)3Si-R'- (Equation I) (In the formula, R is CH3- or CH2CH3-, and, R' is the substrate of the enzyme.

[0009] In some embodiments, R' is [-(CH2)x-(SS)n-(CH2)x-] (In the formula, x is 1 or greater, and, n is 1 or greater.

[0010] In some embodiments, the first dye has an emission peak in the wavelength range of 500 to 1200 nm, and the second dye has an absorption peak in the wavelength range of 500 to 1200 nm.

[0011] In some embodiments, the absorption band, preferably the absorption peak, of the first dye has a shorter wavelength compared to the emission band, preferably the emission peak, of the second dye.

[0012] In some embodiments, the first dye is Absorption peaks in -500~750nm, -Emission peak at least 600nm and Selected to have, The second pigment is, - The absorption spectrum overlapping with the emission spectrum of the first dye, - An emission peak at least 20 nm away from the emission peak of the first dye Selected to have.

[0013] In some embodiments, the emission peak of the second dye has a wavelength of 600 - 950 nm.

[0014] In some embodiments, the first dye and the second dye are each independently selected from the list comprising: - Rhodamine derivatives, - Coumarin derivatives, - Acridine derivatives, - Carbopyronine derivatives, - Oxazine derivatives, - Cyanine dyes, or - Xanthene dyes.

[0015] In some embodiments, the ratio between the first dye and the second dye is equal to the ratio between their respective emission band intensities with a deviation from said ratio within ±20%, preferably within ±15%, preferably within ±10%, preferably within ±5%.

[0016] The present invention further provides a method for preparing a composition for temperature measurement according to the embodiments described herein, comprising: - Immersing unloaded B - PMO particles in a combination of the first dye and the second dye.

[0017] In some embodiments, the method involves applying a lipid bilayer on the surface of the loaded B - PMO particles.

[0018] The present invention further provides a method for obtaining temperature data of a cell, tissue or subject, comprising: - Obtaining emission data of the subject using an optical sensor, wherein the optical sensor is disposed outside the cell, tissue or subject, and - Determining the temperature data of the subject from the emission data, and A method is provided in which a cell, tissue, or object is provided with a temperature measuring composition according to any one of the preceding claims, and a light sensor is configured to detect light emission from the temperature measuring composition.

[0019] In some embodiments, the method involves irradiating cells, tissues, or subjects with light having a wavelength within the absorption band of a first dye.

[0020] In some embodiments, the method is performed in vitro. Legend for the figure [Brief explanation of the drawing]

[0021] [Figure 1] Figure 1 provides a sketch of possible layouts of absorption and emission bands of a first dye and a second dye used in a temperature measuring composition according to an embodiment of the present invention. [Figure 2] Figure 2 shows the various temperature-dependent fluorescence of the first and second dyes. [Modes for carrying out the invention]

[0022] Detailed description of the invention Before describing the systems and methods of the present invention, it should be understood that the present invention is not limited to the specific systems and methods or combinations described, for this reason that such systems and methods and combinations may naturally vary. It should also be understood that the scope of the present invention is limited only by the appended claims, and therefore the terms used herein are not intended to be limiting.

[0023] As used herein, the singular forms "a," "an," and "the" include both singular and plural referents unless otherwise explicitly indicated by the context.

[0024] As used herein, the terms “comprising,” “comprises,” and “comprised of” are synonymous with “including,” “includes,” or “containing,” and are comprehensive or open-ended, not excluding additional unenumerated members, elements, or steps of method. As used herein, the terms “comprising,” “comprises,” and “comprised of” will be understood to include the terms “consisting of,” “consists,” and “consists of.”

[0025] The enumeration of numerical ranges by endpoints includes all numbers and fractions contained within each range, as well as the enumerated endpoints.

[0026] As used herein, the terms “about” or “approximately” when referring to measurable values ​​such as parameters, quantities, and durations mean that variations of a given value and from that value of + / - 10%, preferably + / - 5%, more preferably + / - 1%, and even more preferably + / - 0.1%, are included, insofar as such variations are appropriate for performing the disclosed invention. It should be understood that the values ​​themselves referred to by the modifiers “about” or “approximately” are also specifically and preferably disclosed.

[0027] The terms "one or more" or "at least one," such as one or more members or at least one member of a group of members, are self-explanatory, but to illustrate further, the terms include references to any one of the members, or any two or more of the members, for example, any ≥3, ≥4, ≥5, ≥6, or ≥7 of the members, and at most all of the members.

[0028] All references cited herein are incorporated herein by reference in their entirety. In particular, the teachings of all references specifically mentioned herein are incorporated herein by reference.

[0029] Unless otherwise defined, all terms used in the disclosure of this invention, including technical and scientific terms, have meanings that are generally understood by those skilled in the art to which this invention pertains. Further guidance includes definitions of terms to better understand the teachings of this invention.

[0030] The following sections define different embodiments of the present invention in more detail. Each of these embodiments may be combined with any other embodiment unless otherwise explicitly stated. In particular, any feature indicated as preferred or advantageous may be combined with any other feature indicated as preferred or advantageous.

[0031] Throughout this specification, any reference to “one embodiment” or “embodiment” means that a particular feature, structure, or characteristic described in relation to that embodiment is included in at least one embodiment of the present invention. Therefore, the occurrence of the phrase “in one embodiment” or “in one embodiment” in various places throughout this specification does not necessarily all refer to the same embodiment, but may do so. Furthermore, certain features, structures, or characteristics may be combined in one or more embodiments in any suitable manner, as will be apparent to those skilled in the art from this disclosure. Moreover, some embodiments described herein include some features included in other embodiments, but not others, and at the same time, combinations of features from different embodiments are within the scope of the present invention and form different embodiments, as will be understood to those skilled in the art. For example, in the appended claims, any of the claimed embodiments may be used in any combination.

[0032] In this specification of the present invention, attached drawings are shown only as examples of specific embodiments that form part of this specification and can carry out the invention. The reference numbers in parentheses or bold attached to each element are merely illustrative examples of the elements and are not intended to limit each element. Unless otherwise indicated, all figures and drawings in this specification are not to scale and have been selected for the purpose of illustrating different embodiments of the invention. In particular, the dimensions of various components are given in illustrative terms only, and unless otherwise indicated, the relationships between the dimensions of various components should not be inferred from the drawings.

[0033] It should be understood that structural or logical modifications may be made by utilizing other embodiments without departing from the scope of the present invention. Therefore, the following detailed description should not be construed as restrictive, and the scope of the present invention is defined by the appended claims.

[0034] This specification provides a temperature measurement composition comprising multiple biodegradable periodic mesoporous organic silicas, B-PMO, and particles, where, -The first dye and the second dye are loaded into each B-PMO particle (of multiple B-PMO particles). -Both the first and second dyes are fluorescent. - Preferably, both the first and second dyes are organic, preferably free of heavy metals, and preferably free of metals. -Fluorescence emission from the first dye triggers emission excitation of the second dye, and - The first dye and / or the second dye exhibit temperature-dependent fluorescence.

[0035] Such temperature-measuring compositions may have the advantage of being biodegradable and therefore not accumulating in the subject, making them suitable for in vivo use. Furthermore, temperature-measuring compositions may be free of metals, especially heavy metals, thereby reducing or even eliminating the toxicity of the temperature-measuring composition. Since the two dyes are confined within the B-PMO, the dyes are in close proximity to each other, thereby ensuring efficient energy transfer between the two dyes. This makes it possible for ratiometric thermometers to be based on temperature-measuring compositions. Because two different dyes are used, a high correlation between temperature and fluorescence is possible, enabling the detection of very small temperature differences. Furthermore, because two dyes acting together are used, the signal-to-noise ratio is high and the sensitivity is high. This may allow the use of small amounts of temperature-measuring composition, which is an advantage because the addition of small amounts of temperature-measuring composition does not interfere with the biological process being investigated. The temperature measurement composition allows for further flexibility in the (commercially available) dyes used, thereby enabling the temperature measurement composition to be tailored to desired conditions, such as the assumed temperature range, assumed wavelength, available detection method, available irradiation source, etc. The B-PMO particles used in this invention are particularly easy to synthesize and / or upscale compared to any metal-based nanoparticles. Furthermore, since commercially available dyes may be used, the temperature measurement composition is readily available.

[0036] In some embodiments, the temperature measuring composition is a ratiometric thermometer. In some embodiments, the temperature measuring composition is used as a ratiometric thermometer, preferably in a biological system. As used herein, the term “ratiometric thermometer” means a thermometer whose measurement is based on the intensity change of two emission bands. These ratiometric thermometers may be considered more reliable temperature probes than those based on only a single emission band. In a ratiometric thermometer, the intensity ratio between the two transitions is not impaired by experimental drawbacks such as small material heterogeneity, variations in sensor concentration, or photoelectronic drift of the excitation light source and detector.

[0037] In some embodiments, one of the first or second dyes exhibits temperature-dependent fluorescence, while the other dye exhibits temperature-independent fluorescence. In such embodiments, the fluorescence of the temperature-independent dye can be used as a reference emission signal that can be used for calibration.

[0038] As used herein, the term “periodic mesoporous organosilica” or the abbreviation PMO refers to a type of silica containing organic groups that give rise to a periodic mesoporous structure. PMO may be obtained from organic crosslinked alkoxysilanes by a sol-gel method in the presence of a structure-directing agent, such as a surfactant. The porous framework of PMO can be obtained based on organic functional groups covalently bonding siloxane domains. After synthesis, the structure-directing agent is removed from the pores of the PMO. Generally, PMO can have a well-ordered mesoporous structure, high thermal stability, and / or mechanical stability. Typically, PMO can have a fairly uniform distribution of organic functional groups in the pore walls.

[0039] In some embodiments, the PMO is - A mixture of organically crosslinked alkoxysilanes, or - A mixture of organic crosslinked alkoxysilanes mixed with a silica source such as tetraethyl orthosilicate (TEOS). It may also be made from materials derived from [unspecified material].

[0040] In some embodiments, the mixture may further contain silsesquioxane. As used herein, the term "silsesquioxane" refers to the chemical formula [RSiO 3 / 2 ] n This can refer to organosilicon compounds having (R=H, alkyl, aryl, alkenyl, or alkoxyl). Silsesquioxanes can take the form of cage-like or polymer structures having Si-O-Si bonds and tetrahedral Si vertices, with an inorganic silicate (SiO2) core formed together with an organic outer shell or shell.

[0041] As used herein, the term “biodegradable” refers to the property of a compound or material that is broken down by biological processes, preferably into non-toxic compounds. Preferably, these biological processes occur in the body, preferably in the human body.

[0042] In some embodiments, the biodegradable periodic mesoporous organic silica particles are metabolizable PMO particles, preferably metabolizable by animal or human bodies.

[0043] In some embodiments, the biodegradable periodic mesoporous organic silica particles are removable from animal or human bodies. In some embodiments, the elimination half-life of the biodegradable periodic mesoporous organic silica particles from animal or human bodies is up to 72.0 hours, preferably up to 48.0 hours, preferably up to 24.0 hours, preferably up to 18.0 hours, preferably up to 12.0 hours, and preferably up to 6.0 hours.

[0044] As used herein, the term "fluorescence" refers to the property of a compound, such as a dye, to re-emit light after photoexcitation. The re-emitted light can be any light, such as ultraviolet, visible, or infrared light.

[0045] As used herein, the term “dye” refers to a compound, preferably an organic molecule, having extended conjugation, including a framework of sigma bonds and associated pi systems, that interacts only with specific wavelengths of the light spectrum. Thus, this specific interaction allows the dye to exhibit a particular color. Dyes may differ from organic pigments in that the dye is soluble.

[0046] As used herein, the term "temperature-dependent fluorescence" may refer to a change in the fluorescence behavior of a compound due to the influence of temperature, such that the wavelength of emitted light can be changed under the influence of temperature, the emission intensity can be changed under the influence of temperature, and / or the fluorescence lifetime can be changed under the influence of temperature. Therefore, compounds exhibiting such temperature-dependent fluorescence may be used for emission temperature measurements. Preferably, temperature-dependent fluorescence appears in the visible or infrared portion of the light spectrum.

[0047] In some embodiments, at least one of the first or second dyes is a thermochromic dye, and preferably both the first and second dyes are thermochromic dyes.

[0048] As used herein, the term "thermochromic" refers to the property of a substance to change color in response to a change in temperature.

[0049] As used herein, the term “dye-loaded B-PMO particles” means that the dye is partially or completely impregnated into the B-PMO particles, or that the dye is non-covalently bound to the B-PMO particles. In other words, B-PMO particles may be doped with dye. The dye may be (reversibly) encapsulated in the B-POM pores, or (reversibly) (non-covalently) bound to the walls of the B-PMO particles.

[0050] In some embodiments, the first and second dyes are non-covalently bonded to the B-PMO particles or impregnated into the B-PMO particles.

[0051] In some embodiments, a lipid bilayer is applied to the surface of the loaded B-PMO particles. This lipid bilayer can prevent premature leakage of the dye from the B-PMO particles.

[0052] In some embodiments, the temperature measuring composition is a dispersion in an aqueous phase, preferably an isotonic solution, and preferably a dispersion of B-PMO particles.

[0053] In some embodiments, each B-PMO particle comprises multiple polymer chains of alkoxysilane of formula I.

[0054] -(RO)3Si-R'- (Equation I) (In the formula, R is CH3- or CH2CH3-, and, R' is a substrate for an enzyme, preferably a cellular enzyme, preferably an intracellular enzyme. Since R' is an enzyme substrate, R' can provide biodegradability for B-PMO. Preferably, R' contains at least one disulfide crosslink.

[0055] In some embodiments, R' is [-(CH2) x -(SS) n -(CH2) x -] (In the formula, x is 1 or greater, preferably less than 15, preferably less than 10, preferably less than 5, and, n is 1 or greater, preferably 2 or greater, for example n=2, 4, or 6. Preferably n is up to 10, preferably up to 8, preferably up to 6.

[0056] In some embodiments, 90% of the B-PMO particles have a particle size (d90) of at least 20.0 nm to a maximum of 650.0 nm, preferably at least 30.0 nm to a maximum of 600.0 nm, preferably at least 50.0 nm to a maximum of 500.0 nm, preferably at least 70.0 nm to a maximum of 400.0 nm, preferably at least 100.0 nm to a maximum of 300.0 nm, and preferably at least 150.0 nm to a maximum of 250.0 nm. Such particle sizes are advantageous because the particles are used in the pharmaceutical field and are small enough to reach the desired target, and large enough to hold a sufficient amount of dye.

[0057] In some embodiments, the average wall thickness of B-PMO is at least 5.0 to a maximum of 100.0 nm, preferably at least 7.0 to a maximum of 75.0 nm, preferably at least 10.0 to a maximum of 50.0 nm, and preferably at least 20.0 to a maximum of 40.0 nm. Such wall thickness can be a compromise between rigidity, which provides mechanical stability, and softness and flexibility, which is beneficial for the purpose of delivery and the ability to enter cells.

[0058] In some embodiments, 90% of the pores in the B-PMO have a pore diameter of at least 0.5 nm, preferably at least 0.7 nm, preferably at least 1.0 nm, preferably at least 1.0 nm, and preferably at least 2.0 nm. In some embodiments, 90% of the pores in the B-PMO have a pore diameter of at least 0.5 nm to a maximum of 10.0 nm, preferably at least 0.7 nm to a maximum of 7.0 nm, preferably at least 1.0 nm to a maximum of 5.0 nm, preferably at least 1.0 nm to a maximum of 4.0 nm, and preferably at least 2.0 nm to a maximum of 3.0 nm. Such pore diameters allow for easy uptake of the dye.

[0059] In some embodiments, the BET specific surface area of ​​the B-PMO particles before dye loading is at least 100 m². 2 / g, preferably at least 200m 2 / g, preferably at least 400m 2 / g, preferably at least 600m2 / g, preferably at least 800 m 2 / g, preferably at least 1000 m 2 / g, preferably at least 1200 m 2 / g, preferably at least 1400 m 2 / g, preferably at least 1500 m 2 / g. After loading the dye, these BET specific surface areas can be significantly lower, for example 20 m 2 / g or 50 m 2 / g.

[0060] In some embodiments, the B-PMO particles do not aggregate. In some embodiments, the B-PMO particles may be non-hollow particles. In some embodiments, the B-PMO particles may be hollow B-PMO particles. Hollow particles can be more easily deformed compared to non-hollow particles and thus may have the advantage of being easily taken up by cells.

[0061] In some embodiments, the B-PMO particles are preferably at least the following precursors in the presence of a structure directing agent: - For forming core particles, preferably selected from the list including tetraethyl orthosilicate (TEOS), tetramethyl orthosilicate, tetrapropyl orthosilicate, tetrabutyl orthosilicate, preferably tetraethyl orthosilicate, tetraalkyl orthosilicate, and / or, - A mixture containing: ○ A disulfide silica precursor or a tetrasulfide silica precursor, preferably where the disulfide silica precursor or the tetrasulfide silica precursor is bis(3-trialkoxysilylalkyl) disulfide and / or bis(3-trialkoxysilylalkyl) tetrasulfide, ○In some cases, preferably selected from a list including bis(trialkoxysilyl)ethylene, bis(trialkoxysilyl)methane, bis(trialkoxysilyl)ethane, bis(trialkoxysilyl)ethine, and bis(trialkoxysilyl)benzene, bis(trialkoxysilyl) hydrocarbons Synthesized from, Preferably, the alkoxy group is independently a methoxy group, an ethoxy group, or a propoxy group, Preferably, the alkyl group is independently a methyl group, an ethyl group, a propyl group, or a butyl group. A B-PMO shell is formed around the core particle, or if the core particle is not synthesized, B-PMO particles are formed.

[0062] The ratio of bis(trialkoxysilyl) hydrocarbons to disulfide silica precursors or tetrasulfide silica precursors in the mixture may be modified to achieve a certain biodegradation rate. These precursors have the advantages of not decomposing into toxic compounds, not being too bulky so as to lose the porosity of the PMO, and / or not exhibiting luminescence properties that could interfere with the dye.

[0063] In some embodiments, the volume ratio of bis(trialkoxysilyl) hydrocarbons to the disulfide silica precursor or tetrasulfide silica precursor in the mixture is at least 0.0 to at most 10.0, preferably at least 0.1 to at most 7.5, preferably at least 0.3 to at most 5.0, preferably at least 0.4 to at most 3.0, preferably at least 0.5 to at most 2.5, preferably at least 0.6 to at most 2.0, preferably at least 0.7 to at most 1.5, preferably at least 0.8 to at most 1.3, preferably at least 0.9 to at most 1.1, for example, about 1.0.

[0064] In some embodiments, the core particles are etched away after synthesizing a B-PMO shell around them, preferably using an etching agent such as Na2CO3, NaOH, or polyvinylpyrrolidone (PVP). This can result in hollow B-PMO particles.

[0065] In some embodiments, B-PMO particles are preferably formed in the presence of a structure-directing agent, and contain at least the following precursors: - Mixture containing the following: Preferably, the disulfide silica precursor or tetrasulfide silica precursor is bis(3-trialkoxysilylalkyl)disulfide and / or bis(3-trialkoxysilylalkyl)tetrasulfide, ○In some cases, preferably selected from a list including bis(trialkoxysilyl)ethylene, bis(trialkoxysilyl)methane, bis(trialkoxysilyl)ethane, bis(trialkoxysilyl)ethine, and bis(trialkoxysilyl)benzene, bis(trialkoxysilyl) hydrocarbons Synthesized from, Preferably, the alkoxy group is independently a methoxy group, an ethoxy group, or a propoxy group, Preferably, the alkyl group is independently a methyl group, an ethyl group, a propyl group, or a butyl group. Such B-PMO particles may be non-hollow particles.

[0066] A preferred embodiment of bis(trialkoxysilyl) hydrocarbons is bis(triethoxysilyl)ethylene, which has the formula [A]:

[0067] [ka]

[0068] It can be represented as follows. Bis(triethoxysilyl)ethylene has CAS number 87061-56-1.

[0069] A preferred embodiment of the disulfide silica precursor or tetrasulfide silica precursor is bis(3-triethoxy-silylpropyl) disulfide, which has the formula [B]:

[0070] [ka]

[0071] It can be represented as follows. Bis(3-triethoxysilylpropyl) disulfide has CAS number 56706-10-6.

[0072] A preferred embodiment of the disulfide silica precursor or tetrasulfide silica precursor is bis(3-triethoxy-silylpropyl)tetrasulfide, which has the formula [C]:

[0073] [ka]

[0074] It can be represented as follows. Bis(3-triethoxysilylpropyl)tetrasulfide has CAS number 40372-72-3.

[0075] In some embodiments, B-PMO particles are synthesized from at least the following: - Tetraethyl orthosilicate (TEOS), -Bis(3-triethoxysilylpropyl) disulfide and / or bis(3-triethoxysilylpropyl) tetrasulfide, as well as - In some cases, bis(triethoxysilyl)ethylene.

[0076] In some embodiments, the synthesis of B-PMO can be assisted by the use of a structure-directing agent, preferably a surfactant, preferably cetyltrimethylammonium bromide. The structure-directing agent is preferably removed from the particles after the synthesis of the B-PMO particles.

[0077] In some embodiments, other reagents such as ammonia, ethanol, distilled water, HCl, and / or Na2CO3 may be used in the synthesis of B-PMO particles.

[0078] In some embodiments, hollow B-PMO particles are prepared by a multi-step synthesis, in which, -In the first step, an SiO2 core is prepared, preferably starting from tetraalkyl orthosilicate, in the presence of an aqueous base such as aqueous ammonia. -In the second step, preferably in an aqueous solvent, preferably the SiO2 core is brought into contact with a structure-directing agent and the mixture described herein to provide a disulfide-based organic shell around the SiO2 core, -In the third step, preferably the shell-core structure is brought into contact with an acidifying alcohol, such as acidifying ethanol, to remove the structure-directing agent from the shell-core structure obtained in the second step. -In the fourth step, the SiO2 core is etched away using an etching agent.

[0079] Unless otherwise stated, the wavelengths (for emission bands, emission peaks, absorption bands, and absorption peaks) referred herein for the dyes are present within the B-PMO particles, which are suspended in an aqueous environment (preferably at a temperature midway through the target temperature range). Those skilled in the art will recognize that the emission bands, emission peaks, absorption bands, and absorption peaks of dyes in an organic environment may differ from those in an aqueous environment. When selecting dyes to be included in B-PMO, those skilled in the art can make selections based on reported values ​​in an aqueous environment. Alternatively or additionally, one or more experiments may be performed to confirm the wavelengths of the emission bands, emission peaks, absorption bands, and absorption peaks of dyes in B-PMO suspended in an aqueous environment to confirm the dyes selected to be included in the embodiments of the present invention.

[0080] In some embodiments, the first dye has an emission peak in the wavelength range of 500 to 1200 nm, and the second dye has an absorption peak in the wavelength range of 500 to 1200 nm.

[0081] In some embodiments, both the first and second dyes have both absorption and emission peaks within the wavelength range of 500 nm to 1200 nm.

[0082] In some embodiments, the first dye has an emission peak that partially overlaps with the absorption peak of the second dye.

[0083] In some embodiments, the absorption band, preferably the absorption peak, of the first dye has a shorter wavelength compared to the emission band, preferably the emission peak, of the second dye.

[0084] In some embodiments, the emission band, preferably the emission peak, of the first dye has a shorter wavelength compared to the emission band, preferably the emission peak, of the second dye.

[0085] Preferably, the first and second dyes are different, preferably have different chemical structures, and preferably have different absorption and / or emission spectra. Preferably, the first and second dyes are photoluminescent dyes.

[0086] In some embodiments, the first dye and the second dye are: -Water stability means that when dissolved in water, the absorption and emission spectra of the dye do not change over time. - pH stability means that the absorption and emission spectra of the dye do not change when exposed to different pH values. - Low toxicity, which can be determined by cell viability assays. - In particular, the dye does not decompose in a temperature range of at least 20°C to a maximum of 60°C, preferably at least 25°C to a maximum of 50°C, and is thermally stable over that temperature range. -Photostable, meaning that the absorption and emission spectra of the dye do not change after light irradiation, and / or, - High quantum yield.

[0087] In some embodiments, the first dye is suspended in an aqueous solvent (preferably within B-PMO) at a temperature midway through the desired temperature range of the assumed temperature measurement composition, below: Absorption peaks in -500~750nm, - Preferably, an emission peak in the far-red to near-infrared spectrum at least 600 nm, preferably 600 nm to 800 nm, preferably 650 nm to 800 nm. Selected to have.

[0088] On the other hand, the second dye is suspended in an aqueous solvent (preferably within B-PMO) at a temperature midway through the desired temperature range of the assumed temperature measurement composition, below: - The absorption spectrum overlapping with the emission spectrum of the first dye, - An emission peak located at least 20 nm, preferably at least 40 nm, preferably at least 60 nm, preferably at least 80 nm, preferably at least 90 nm away from the emission peak of the first dye Selected to have.

[0089] An example of such a first pigment is Rh800®, which in an aquatic environment, An absorption spectrum in the range of -550 to 720 nm, having a maximum value at 680 nm. - Emission spectrum with peak at 705 nm, from 650 to 850 nm. It has, This is combined with a second pigment, ICG®, which, in an aqueous environment, Absorption spectrum from -600 to 750 nm, -815nm emission peak This system is preferably excited at a wavelength of 624 nm or 660 nm.

[0090] Preferably, the temperature measuring composition is excited by irradiation with light having a wavelength of ±50 nm, preferably ±35 nm, preferably ±25 nm, preferably ±20 nm, preferably ±15 nm, preferably ±10 nm, preferably ±5 nm, preferably ±2 nm, which is equal to the peak of the absorption spectrum of the first dye.

[0091] In some embodiments, the emission peak of the second dye has a wavelength of 500 to 2100 nm. This allows for the detection of synchrotron radiation using existing technologies. Preferably, the emission peak of the second dye has a wavelength of 500 to 900 nm. This allows for the detection of synchrotron radiation using conventional CCD cameras.

[0092] In some embodiments, the emission peak of the second dye has a wavelength of 600–950 nm. This is a “first biological window” that can allow light to penetrate and diffuse into tissue. Both emission in the infrared spectrum or emission through the biological window can be difficult to achieve with lanthanide nanomaterials (prior art).

[0093] In some embodiments, the emission peak of the second pigment has a wavelength of 780–1400 nm, which can allow light to travel through the skin.

[0094] In some embodiments, the emission peak of the second dye has a wavelength of 500 to 2100 nm, preferably 500 to 1000 nm, preferably 500 to 900 nm, preferably 600 to 900 nm, and preferably 780 to 900 nm.

[0095] In some embodiments, both the first and second dyes are water-soluble. In some embodiments, the first and second dyes are dissolved in a solvent and encapsulated in B-PMO, the solvent being preferably an alcohol, preferably ethanol.

[0096] In some embodiments, the first and second dyes are not covalently bonded together.

[0097] In some embodiments, the ratio between the first and second dyes depends on the intensity of their respective emission bands. The ratio between the first and second dyes is equal to the ratio between their respective emission band intensities, with a deviation from that ratio of ±20%, preferably ±15%, preferably ±10%, and preferably ±5%. Such a ratio has the advantage that the emission of one dye does not render the emission of the other dye undetectable. Therefore, preferably, the emission of both the first and second dyes is distinguishable from the noise level.

[0098] In some embodiments, the molar ratio of the first dye to the second dye is 1 / 99 to 99 / 1, preferably 5 / 95 to 95 / 5, preferably 10 / 90 to 90 / 10, preferably 25 / 75 to 75 / 25, and preferably about 50 / 50. Such ratios have the advantage that the emission of one dye does not render the emission of the other dye undetectable. Therefore, preferably, the emission of both the first and second dyes is distinguishable from the noise level.

[0099] In some embodiments, the absorption wavelength band peak of the first dye differs from the emission wavelength band peak of the second dye, preferably having a different height and more preferably a different wavelength range.

[0100] In some embodiments, the first dye and / or the second dye are fluorescent dyes, preferably meaning that luminescence by the dye ceases after the excitation of the dye stops.

[0101] Throughout this specification, the following pigments are referred to: -Rh800(registered trademark), xanthene dye, CAS 137993-41-0 (can be represented by formula [X]), -Atto610(registered trademark), carvopyronine derivative, CAS 303952-45-6 (may be represented by formula [XI]), -IR806(registered trademark), cyanine dye, CAS76783-59-0 (can be represented by formula [XII]), and, -ICG®, or indocyanine green, cyanine dye, CAS 3599-32-4 (may be represented by formula [XIII]).

[0102] [ka]

[0103] [ka]

[0104] Other dyes disclosed herein are as follows:

[0105] [Table 1]

[0106] In some embodiments, the first dye and the second dye can be independently: -Rhodamine derivatives, - Coumarin derivatives, - Acridine derivatives, -Carbopyronine derivatives, -Oxazine derivatives, - Cyanine pigment, or - Xanthene pigment.

[0107] In this field, several dyes derived from rhodamine, coumarin, acridine, carbopyronine, or oxazine are sold by Atto-Tec, and are therefore called Atto dyes, and are particularly suitable for use as a primary or secondary dye.

[0108] In some embodiments, the first dye is a xanthene dye or a derivative of rhodamine, coumarin, acridine, carbopyronine, or oxazine, while the second dye is a cyanine dye.

[0109] In some embodiments, the first dye is a derivative of rhodamine, coumarin, acridine, carbopyronine, or oxazine, while the second dye is a cyanine dye. In some embodiments, the first dye is a derivative of carbopyronine, while the second dye is a cyanine dye, for example, Atto610® as the first dye and IR806® as the second dye.

[0110] In some embodiments, the first dye is a xanthene dye, while the second dye is a cyanine dye, such as Rh800® as the first dye and ICG® as the second dye.

[0111] In some embodiments, the combination of the first and second dyes is selected from the following pairs:

[0112] [Table 2]

[0113] Based on the teachings herein, those skilled in the art can select other suitable combinations of the first and second dyes disclosed herein, and therefore the present invention is not limited to the dyes disclosed herein.

[0114] In some embodiments, the temperature measuring composition is - 1.0 mol% Atto610 (registered trademark) as the first dye and 99.0 mol% IR806 (registered trademark) as the second dye. - 1.5 mol% Atto610 (registered trademark) as the first dye and 98.5 mol% IR806 (registered trademark) as the second dye. - 50.0 mol% Rh800 (registered trademark) as the first dye and 50.0 mol% ICG (registered trademark) as the second dye. - 25.0 mol% Rh800 (registered trademark) as the first dye and 75.0 mol% ICG (registered trademark) as the second dye, or -Contains 75.0 mol% Rh800 (registered trademark) as the first dye and 25.0 mol% ICG (registered trademark) as the second dye. Here, mol% is expressed in comparison to the total number of moles of the dye mixture in the temperature measurement composition.

[0115] A method for preparing a temperature measuring composition according to embodiments disclosed herein, -The present specification provides a method comprising immersing unloaded B-PMO particles in a combination of a first dye and a second dye, preferably as a solution, more preferably as an aqueous solution.

[0116] In some embodiments, ultrasound is used during immersion. This ensures good dispersion of B-PMO particles in the solution.

[0117] In some embodiments, the B-PMO particles and the solutions of the first and second dyes are stirred, preferably agitated, during immersion.

[0118] In some embodiments, the immersion lasts for at least 1 hour to a maximum of 72 hours, preferably at least 2 hours to a maximum of 36 hours, preferably at least 6 hours to a maximum of 48 hours, and preferably at least 12 hours to a maximum of 24 hours.

[0119] In some embodiments, the loaded B-PMO particles can be recovered from the solution by filtration, preferably by centrifugation.

[0120] In some embodiments, the recovered, loaded B-PMO particles are preferably washed with water. In some embodiments, the recovered, loaded B-PMO particles or the washed, loaded B-PMO particles may be dried.

[0121] In some embodiments, the method involves applying a lipid bilayer to the surface of the loaded B-PMO particles. This lipid bilayer can prevent the leakage of dye from the B-PMO particles and can improve the dispersibility of the particles in aqueous solution. Preferably, the lipid bilayer is a 1,1-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayer. Preferably, the loaded B-PMO particles are brought into contact with a lipid solution in an aqueous alcohol (preferably ethanol) solution. Ultrasound may be applied during this contact step. The coated and loaded B-PMO particles may be recovered by filtration, preferably by centrifugation, and optionally dried.

[0122] A method for obtaining temperature data of cells, tissues, or objects, - Acquiring light emission data of a target using a light sensor, wherein the light sensor is located outside the cell, tissue, or target. - Determining the target temperature data from the emission data, Includes, A method is provided in which a temperature measuring composition according to embodiments disclosed herein is provided for cells, tissues, or subjects, and a light sensor is configured to detect light emission from the temperature measuring composition.

[0123] In some embodiments, the emission is infrared light. This may have the advantage that this data can penetrate skin or tissue material, and temperature data can be obtained from within the tissue or object.

[0124] In some embodiments, the method involves irradiating cells, tissues, or subjects with light having wavelengths within the absorption band of a first dye, preferably light having wavelengths equal to the peak of the absorption spectrum of the first dye ±50 nm, preferably ±35 nm, preferably ±25 nm, preferably ±20 nm, preferably ±15 nm, preferably ±10 nm, preferably ±5 nm, preferably ±2 nm. Preferably, the light having wavelengths within the absorption band of the first dye is infrared light. This may have the advantage of being able to penetrate deeply into tissues and skin.

[0125] In some embodiments, the method is performed in vitro, preferably to obtain temperature data of cells and tissues.

[0126] In some embodiments, the light sensor may also be part of a spectrofluorometer, which may also include a light source for illuminating cells, tissues, or objects.

[0127] In some embodiments, the light sensor includes an imaging sensor such as a CCD camera. In some embodiments, the thermochromic composition is delivered to the target by intravenous injection.

[0128] In some embodiments, the temperature data is targeted to the area beneath the skin. In some embodiments, the target is a (benign or malignant) tumor or a temperature abnormality resulting from another health condition, such as an inflammatory condition or arthritis. Preferably, the target is related to melanoma, breast cancer, head cancer, and / or neck cancer. These cancers can be particularly selected as targets because they do not grow too deep below the shin and are well within the probing range of the temperature measuring compositions according to the embodiments disclosed herein.

[0129] In some embodiments, determining the temperature data of a target from emission data is done via a calibration curve, preferably a calibration curve specific to the temperature measuring composition used. In some embodiments, the calibration curve is a curve of the wavelength of the emission peak of a second dye as a function of temperature.

[0130] The present invention is included solely for illustrative purposes of specific aspects and embodiments of the invention and will be more readily understood by referring to the following examples, which are not intended to limit the invention.

[0131] method The emission spectrum and peaks of the dye are recorded using a spectrofluorometer. The fluorescence emission spectrum shows the change in fluorescence intensity as a function of the emission wavelength. For measurement, the excitation wavelength is set to the wavelength of known absorption by the sample (usually the strongest absorption point is selected), and the wavelength of the emission monochromator is scanned over the desired emission range to record the fluorescence intensity as a function of the emission wavelength to the detector.

[0132] For all measurements here, an Edinburgh Instruments FLS980 spectrofluorometer was used.

[0133] The absorption spectra and peaks of the dye are recorded using a spectrophotometer. The absorption spectrum shows the change in the absorbance of the sample as a function of the wavelength of the incident light. The absorption spectrum is measured by varying the wavelength of the incident light using a monochromator and recording the intensity of the transmitted light to the detector. The intensity of the light transmitted through the sample is "I 試料 (Analytes dissolved in a solvent, etc.) and the intensity of light transmitted through the blank "I ブランク Record (solvent only) and A=log10(I ブランク / I 試料 The absorbance of the sample is calculated using ).

[0134] The excitation spectrum and peaks of the dye are recorded using a spectrofluorometer. The fluorescence excitation spectrum shows the change in fluorescence intensity as a function of the excitation light wavelength. The wavelength of the emission monochromator is set to the wavelength of known fluorescence emission from the sample (usually the strongest emission intensity), and the wavelength of the excitation monochromator is scanned over the desired excitation range to record the fluorescence intensity as a function of the excitation wavelength to the detector.

[0135] The particle size of the B-PMO particles here is determined using transmission electron microscopy (TEM). Alternatively, scanning electron microscopy (SEM) may also be used.

[0136] The sizes of multiple particles are measured using image processing software such as ImageJ or an equivalent. Typically, at least 50 to 100 particles are evaluated from different areas of a TEM grid. This is then plotted, usually in the form of a histogram, to show the average size and the variation from the average size.

[0137] The wall thickness of B-PMO is evaluated using transmission electron microscopy (TEM). Multiple images are taken, and then the wall thickness of multiple B-PMO particles (at least 50-100) is measured and averaged using visualization software (ImageJ or equivalent). [Examples]

[0138] Examples Example 1: Preparation of hollow B-PMO particles In this example, hollow B-PMO particles were synthesized in a three-step procedure. First, SiO2 template particles were prepared. For this purpose, 15 mL of aqueous ammonia (25%) was added to 135 mL of ethanol and 2 mL of distilled water. The mixture was stirred for 5 minutes, then 6 mL of tetraethyl orthosilicate (TEOS) was added dropwise, and the mixture was stirred overnight at room temperature. In the second step, to obtain disulfide-based B-PMO@SiO2 core-shell particles, 100 mL of the as-prepared SiO2 suspension was taken, placed in a new flask, and further diluted with 200 mL of distilled water. 30 mL of 0.12 M cetyltrimethylammonium bromide (CTABr) solution was added to the SiO2 suspension, and the flask was stirred for 30 minutes. Next, a mixture of 400 μL of bis(triethoxysilyl)ethylene precursor and 400 μL of bis(3-triethoxysilylpropyl) disulfide precursor was added dropwise and stirred for 24 hours (the flask was left at room temperature). The following day, the resulting powder was collected by centrifugation and washed three times with water. The powder was then dried in an oven at 80°C. CTABr was extracted from the PMO pores by adding the powder to an acidified ethanol solution (1 mL of 37% HCl in 100 mL of ethanol) under reflux, and this process was repeated three times. The resulting mixture was again centrifuged, washed with water, and left to dry in an oven at 80°C. Finally, in the third step, to obtain the final material, 0.1 g of dried B-PMO@SiO2 powder was placed in a flask containing 0.636 g of Na2CO3 and 20 mL of distilled water. This mixture was heated at 80°C for 1.5 hours. The suspension was centrifuged, washed twice with water, and dried in a drying oven at 80°C. The resulting material is referred to as B-PMO particles.

[0139] Example 2 Loading of the first and second dyes into hollow B-PMO particles 15 mg of the as-prepared dried B-PMO particles from Example 1 were weighed and dispersed in 3 mL of distilled water containing a dissolved dye mixture (the ratio of specific dyes varied, but as a standard, the total weight of the two dyes combined was 2.5 mg). When poorly water-soluble dyes were used, other solvents, such as a water:ethanol mixture, could be used in the loading process. The ratio of dyes depended on the dyes used and was related to their luminescence intensity.

[0140] An ultrasonic bath was used for 5 minutes to ensure proper dispersion of B-PMO in the solvent. The mixture was stirred in the dark at room temperature for 24 hours (the flask was covered with aluminum foil to prevent dye degradation). The stirring speed was set to low (200-300 rpm).

[0141] Next, the solid particles were separated by centrifugation, gently washed with water, and then subjected to further centrifugation. The centrifugal speed was set to 4500 rpm and the centrifugation time was set to 5 minutes as a standard (the centrifugation time may be extended if there are problems with sedimentation). The product was dried in an oven at 80°C.

[0142] In the final step, a lipid bilayer coating was constructed on the dye-loaded B-PMO particles to reduce the possibility of dye leakage and to improve the dispersibility of the particles in aqueous solution. For this purpose, a solution of 1,1-dioleoyl-sn-glycero-3-phosphocholine (DOPC) was first prepared (28 mg of DOPC dissolved in 6 mL of distilled water and 4 mL of ethanol). For every 5 mg of dye-loaded B-PMO dry powder, 500 μL of the DOPC solution was taken and added to a conical centrifuge tube. The mixture was dispersed using ultrasound (2-3 minutes). 4.5 mL of distilled water was added, and the mixture was immediately shaken vigorously (manually or using a vortex). The mixture was centrifuged (4500 rpm, 5 minutes) and redispersed twice in water for purification (redispersion in water using ultrasound and continuous centrifugation). The final product was dried in an oven at 80°C and stored in the dark in powder form.

[0143] Example 3: Optimization of the ratio of the first pigment to the second pigment The selection of the dye ratio is performed by experimental optimization. A known molar amount (0.005 mmol) of the first dye is encapsulated in 15 mg of B-PMO (from Example 1), and a lipid bilayer is constructed around it for water stability and to prevent dye leakage. The absorption spectrum and excitation and emission spectra are then recorded after redispersion in distilled water. The same molar amount of the second dye is separately encapsulated in B-PMO, and the lipid bilayer is wrapped around the particle. The absorption spectrum and excitation and emission spectra are then recorded.

[0144] Based on this first data, the excitation and emission ranges and brightness of the first and second dyes are evaluated in a B-PMO environment and in the presence of water quenching. Depending on the performance of the dyes in this environment, different ratios of the first and second dyes are proposed. For example, if both the first and second dyes exhibit similar intensity in a B-PMO environment, ratios of 25:75, 50:50, and 75:25 are investigated to identify the highest sensitivity (Sr). In the synthesis of the mixed dye using 15 mg of B-PMO (Example 1), the target total amount of dye is 2.5 mg. If one of the dyes exhibits a much stronger emission intensity compared to the other, ratios such as 1:99, 5:95, and 10:90 are starting points for identifying the highest sensitivity. Feedback from experiments may lead to further optimization of the dye ratios.

[0145] Example 4: Layout of absorption and emission bands of the first and second dyes Figure 1 provides a sketch of possible layouts of absorption and emission bands of a first and second dye used in a temperature measuring composition according to an embodiment of the present invention. The absorption band (A1) of the first dye is located at a shorter wavelength and reaches an absorption peak (A1p). The emission band (E1) of the first dye is shifted to a longer wavelength compared to A1 and reaches an emission peak (E1p). At even longer wavelengths, an absorption band (A2) of the second dye can be found, reaching an absorption peak (A2p). Since the absorption band (A2) of the second dye overlaps (O) with the emission band (E1), emission from the first dye can excite the second dye. Finally, the emission band (E2) of the second dye is located at the upper end of the wavelength and reaches an emission peak (E2p). Preferably, the temperature measuring composition containing the first and second dyes should be irradiated with light having a wavelength of A1p, and the response should be monitored preferably around E2p.

[0146] Example 5: Temperature-dependent fluorescence. Figure 2 shows the various temperature-dependent fluorescence of the first and second dyes. Figure 2A shows the emission bands (I1) of the first dye and (I2) of the second dye. As the temperature increases, the fluorescence intensity of the first dye decreases, but the fluorescence of the second dye also decreases.

[0147] Figure 2B shows the emission bands (I1) of the first dye and (I2) of the second dye. As the temperature increases, the fluorescence intensity of the first dye decreases, while the fluorescence of the second dye remains constant.

[0148] Figure 2C shows the emission bands (I1) of the first dye and (I2) of the second dye. As the temperature increases, the fluorescence intensity of the first dye decreases, while the fluorescence of the second dye increases.

[0149] Comparative Examples and Examples As shown in Table 3, various conventional temperature measuring compositions (comparative examples) and temperature measuring compositions according to embodiments of the present invention (examples) were tested. In the examples, B-PMO prepared in Example 1 was used, and the dyes shown in Table 3 were loaded according to Example 2. In all examples in Table 3, the fluorescence of the temperature measuring compositions was investigated over the temperature range shown in the table. The emission range was determined according to the temperature within the indicated temperature range.

[0150] The relative sensitivity (Sr) was determined for each temperature measurement composition. Relative sensitivity S r This shows the relative change in temperature measurement parameters for each degree of temperature change, %℃ -1 It is expressed as follows. This is calculated using the following formula.

[0151]

number

[0152] During the ceremony,

[0153]

number

[0154] Here, I1 and I2 are the maximum peak intensity (or integrated area under the peak) at the selected wavelength.

[0155] For each temperature measurement composition, the temperature uncertainty or resolution (δT) was determined based on the signal-to-noise ratio.

[0156] The temperature uncertainty is calculated using the following formula.

[0157]

number

[0158] [Table 3]

Claims

1. A temperature measuring composition comprising multiple biodegradable periodic mesoporous organic silicas, B-PMO, and particles, - Each B-PMO particle is loaded with the first dye and the second dye. - Both the first dye and the second dye are fluorescent, - The fluorescence emission from the first dye triggers emission excitation of the second dye, - The first dye and / or the second dye exhibit temperature-dependent fluorescence, Composition for temperature measurement.

2. The temperature measuring composition according to claim 1, wherein both the first dye and the second dye are organic.

3. The temperature measuring composition according to claim 1 or claim 2, wherein each B-PMO particle comprises multiple polymer chains of alkoxysilane of formula I: -(EN) ３ Si-R'- (Equation I) (In the formula, R is CH ３ - or CH ２ CH ３ - and, R' is the substrate of the enzyme.

4. R' is [-(CH2)x-(S-S)n-(CH2)x-] (In the formula, x is 1 or greater, and, n is 1 or greater. The temperature measuring composition according to claim 3.

5. The temperature measuring composition according to any one of the preceding claims, wherein the first dye has an emission peak in the wavelength range of 500 to 1200 nm, and the second dye has an absorption peak in the wavelength range of 500 to 1200 nm.

6. The temperature measuring composition according to any one of the preceding claims, wherein the absorption band, preferably the absorption peak, of the first dye has a shorter wavelength than the emission band, preferably the emission peak, of the second dye.

7. The first pigment described above, Absorption peaks in -500 to 750 nm, - Emission peak at least 600 nm and Selected to have, The second dye described above, - The absorption spectrum overlapping with the emission spectrum of the first dye, - An emission peak at least 20 nm away from the emission peak of the first dye A temperature measuring composition according to any one of the preceding claims, selected to have the following characteristics.

8. The temperature measuring composition according to any one of the preceding claims, wherein the emission peak of the second dye has a wavelength of 600 to 950 nm.

9. The first dye and the second dye are independent of each other. - Rhodamine derivatives, - Coumarin derivatives, - Acridine derivatives, -Carbopyronine derivatives, -Oxazine derivatives, - Cyanine pigment, or - Xanthene pigments A temperature measuring composition according to any one of the preceding claims, selected from the list including the following:

10. The temperature measuring composition according to any one of the preceding claims, wherein the ratio between the first dye and the second dye is equal to the ratio between their respective emission band intensities, with a deviation from said ratio of within ±20%, preferably within ±15%, preferably within ±10%, and preferably within ±5%.

11. A method for preparing a temperature measuring composition according to any one of the preceding claims, - A method comprising immersing unloaded B-PMO particles in a combination of the first dye and the second dye.

12. The method according to claim 11, further comprising applying a lipid bilayer to the surface of the loaded B-PMO particles.

13. A method for obtaining temperature data of cells, tissues, or objects, - Acquiring light emission data of the target using a light sensor, wherein the light sensor is located outside the cell, tissue, or target. - Determining the temperature data of the target from the aforementioned light emission data, Includes, A method wherein the cell, tissue, or object is provided with the temperature measuring composition according to any one of the preceding claims, and the light sensor is configured to detect light emission from the temperature measuring composition.

14. The method according to claim 13, comprising irradiating the cells, tissue, or target with light having a wavelength within the absorption band of the first dye.

15. The method according to claim 13 or claim 14, performed in vitro.

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