JP2014226163A - Composite-type optical fiber equipped with functional probe - Google Patents

Abstract

PROBLEM TO BE SOLVED: To solve such a problem that it is difficult to obtain high spatial resolution while an outside diameter maintains 1 mm or 1 mm or less in an existing optical fiber, and further there exists no such a multifunctional diagnostic device as enabling biofunction observation while having an ultrafine diameter.SOLUTION: A composite-type extra-fine diameter optical fiber bundle is capable of observing a visible light and a fluorescent light at the same time, and has a high spatial resolution capable of observing cells. The irradiation using a laser and an LED light source is made possible along with the fluorescent light observation at the same time. An environment-responsive sensor is equipped on a tip part. An ultrafine form is maintained by optimizing a fiber component and bundle structure, and holds multi-functions. Vital observation and function measurement are made possible by mounting a functional probe on a fiber tip. The functional probe seals a low molecular substance in nanoparticles, and bonds it to fibers.

Description

  The present invention relates to a device for measuring / analyzing in-vivo temperature and the like, diagnosing an affected area, and a technique for manufacturing the device.

  Various techniques such as endoscopy, X-ray photography, CT examination, and ultrasonic examination have been put to practical use as methods for obtaining biological information. Of these, endoscopic technology is one of the most important measurement methods that can be applied to various parts of humans. Various diagnostics and treatments that do not impose a burden on the patient by installing various functions at the distal end of the endoscope. The technology can be established and has a wide range of applications such as robotic surgery. Furthermore, diagnosis and treatment under MRI has the advantage of reducing the burden on the patient and monitoring the dynamic situation in real time during the operation, which can greatly contribute to improving the quality of diagnosis and treatment. However, with regard to endoscopes, the wall with a 1.7 mm outer diameter cannot be easily exceeded, and the spatial resolution of current ultra-thin endoscopes (around 2 mm outer diameter) and other noninvasive diagnostic methods is always sufficient. In addition, there is a demand for improvement in the accuracy of identifying a lesion, and establishment of a diagnosis and treatment method for diagnosis and treatment at the cellular level.

  Body temperature measurement is a basic technique for knowing the state of a living body that has been used for a long time as one of vital signs. Although it has been scientifically proven that there are temperature changes in many reactions at the cellular molecular level, temperature measurement at the cellular level is useful for diagnosis and treatment, and is applied to biological function measurement using environmental responsiveness. It has not reached.

  In the present invention, among the environmentally responsive dyes, the temperature sensitive dye is encapsulated (encapsulated) in an organic polymer, and the encapsulated dye is fixed to the tip of the optical fiber, so that the temperature at the contact site is increased. Measurement is possible, and for example, the temperature of the brain surface or deep part in the living body can be measured. Temperature range and sensitivity can be controlled by changing the type of organic polymer. Sensitivity and color tone can be changed by changing the type of chromophore. When this measuring instrument can detect temperature changes below 0.1 ° C, the structure of tissues and cells can be visualized through a temperature map. In addition, it is possible to measure neural activity in the cranial nervous system.

  The composite optical fiber device equipped with the function of sensing biological tissue information according to the present invention is characterized in that an optical fiber in which an organic polymer containing an environmentally responsive dye is immobilized at the tip is used as a part. Yes. In vivo information given to the environmentally responsive dye can be analyzed through an optical fiber.

1) Environmentally Responsive Dye As the environmentally responsive dye according to claims 1 to 5, for example, a temperature sensitive fluorescent dye, a pressure sensitive fluorescent dye, an oxygen sensitive fluorescent dye, a metal (metal ion) sensitive fluorescent dye, pH (acid, Base) sensitive fluorescent dyes and the like. A temperature sensitive fluorescent dye will be described as a representative example.

[Temperature sensitive fluorescent dye]
Examples of the temperature-sensitive fluorescent dye used in the present invention include compounds whose fluorescence intensity changes linearly with respect to temperature. Preferably, a substance exhibiting a change in fluorescence intensity of 0.1% / 1 ° C. or more, more preferably 1.0% / 1 ° C. or more in a predetermined temperature range is preferable. It is preferably selected from europium (Eu), lanthanum (La), samarium (Sm), gadolinium (Gd), terbium (Tb), dysprosium (Dy), thulium (Tm), yttrium (Yb), and lutetium (Lu). A complex compound of a rare earth element and a β-diketone chelate or a complex compound of iridium (Ir) and an aromatic is used. Among these, a complex compound of europium and β-diketone chelates and a complex compound of iridium (Ir) and aromatics are preferable. Specifically, tris (thenoyltrifluoroacetonato) europium (III) (Eu (III) (TTA) ₃ ) or a derivative thereof (such as a hydrate), tris (benzoylacetonato) europium (III) (Eu ( III) (bac) ₃ ), tris (dibenzoylmethanato) europium (III) (Eu (III) (dbm) ₃ ), tris (hexafluoroacetylacetonato) europium (III) (Eu
(III) (hfacac) ₃ ), tris (acetylacetonato) europium (III) (Eu (III) (acac) ₃ ), (1,10-phenanthroline) tris (thenoyltrifluoroacetate) europium (III) (Eu (III) (TTA) ₃ PHEN), tris (2-phenylpyridine) iridium (III) (Ir (III) (ppy) ₃ ), and particularly because the yield of fluorescence near room temperature is high. Tris (thenoyltrifluoroacetonate) europium (III) (Eu (III) (TTA) ₃ ) or a derivative thereof (such as a hydrate) is most preferably used.

  When the temperature-sensitive fluorescent dye-containing copolymer particles are encapsulated and fixed with a difference in concentration, the fluorescence intensity varies and an error occurs as a temperature detection element. The variation can be corrected by simultaneously enclosing a non-temperature sensitive fluorescent dye as a reference. The non-temperature sensitive fluorescent dye is preferably not overlapped with the fluorescence wavelength distribution of the temperature sensitive fluorescent dye used. When the Eu complex is excited at a wavelength near 300 nm, energy emission near 600 nm (red light) is observed. In this case, the reference is preferably a fluorescent dye whose color is observed at 500 nm or less (blue to green light), and examples thereof include fluorescein, its derivatives, rhodamine derivatives, cyanine derivatives, and the like. Specifically, fluorescein isothiocyanate (FITC). , Rhodamine B and the like.

2) Organic polymer Preferred examples of the organic polymer according to claim 2 include, for example, Poly (ethylene); Poly (propylene); Poly (ethylene-co-propylene); Poly (ethylene-co-butene); Poly ( (ethylene-co-hexene); Poly (ethylene-co-octene); Poly (4-methyl-1-pentene); Poly (acrylic acid); Poly (acrylamide); amphiphilic derivative of dextran; Poly (acrylonitrile); Poly ( butyl acrylate); Poly (n-butyl cyanoacrylate); Poly (butyl methacrylate); Poly (methacrylic acid); polymeric surfactant based on carboxymethyl cellulose and alkyl poly (etheroxy) acrylate; Carboxymethylated poly (ethylene glycol); Poly (chloromethyl styrene) Poly (styrene); Poly (methyl methacrylate); Poly (isobutyl methacrylate); Poly (dodecyl methacrylate); Poly ((dimethylamino) ethyl methacrylate); Poly (ethyl cyanoacrylate); Poly (ethylene glycol); Poly (glycidyl methacrylate) Poly (hydroxylethyl methacrylate); Poly (hexyl methacrylate); Poly (ethylene-co-butylene); Poly (ethylene-co-butylene) -b-poly (ethylene oxide); Poly (lauryl methacrylate) ); monomethoxy-poly (ethylene glycol); monomethoxy-poly (ethylene oxide) -poly (lactic acid); Poly (N-methylolacrylamide); Poly (N-vinyl pyrrolidone); Poly (oligo (ethylene oxide) monomethyl ether methacrylate) ; Pullulan acetate; Polyaniline-poly (styrenesulfonic acid); Poly (γ-benzyl-l-glutamate) -b-poly (ethylene oxide); Poly (ε-caprolactum); Poly (N, N-dimethylacrylamide) with a reactive trithiocarbonate Poly (oxyethylene) -poly (oxypropylene) copolymer; Poly (hydroxyl butyrate); Poly (heptadecafluorodecylacrylate); Poly (hydroxyethyl methacrylate); Poly (lactide-fumarate); Poly (d, l-lactic acid-co-glycolic acid) Poly (lactide-co-glycolide fumarate); Poly (l-lactic acid); Poly (α, β-l-malic acid); Poly (methacrylic acid-co-styrene); Poly (N-isopropylacrylamide); Poly ( N-isopropylacrylamide-co-methacrylic acid); Poly (ethylene oxide) -poly (propylene oxide) ethylene diamine co-polymer; Poly (organophosphazene); Poly [2- (3-thienyl) acetyl-3,3,4,4 , 5,5,6,6,7,7,8,8,8-tridecafluorooctanoate]; Poly (styrenesulfonic aci d); Poly (trimethylene carbonate); Poly (vinyl alcohol); SHOA, Poly (stearyl methacrylate); 5-sulfoisophthalic acid dimethyl ester sodium salt modified tetracarboxylic acid-terminated polyester; Poly (vinyl acetate) It is not limited.

3) Binder In the present invention, preferred examples of the binder according to claim 5 include organic polymer compounds, inorganic oxides such as silicon oxide, silicon-based polymer compounds, and the like, but are not limited thereto. .

  As a method for immobilizing organic polymer fine particles in which a stimulus-responsive dye is encapsulated in the fiber tip using a binder, the organic polymer fine particles in which a binder or a precursor thereof and an environment-responsive dye are encapsulated are dissolved or dispersed. There is a method in which a liquid is applied to the fiber tip and dried and solidified.

  Nanoparticles immobilized with the outer diameter maintained at 1 mm could visualize the structure of brain tissue and tract structure. By inserting into the deep brain, the brain structure can be easily visualized, and at the same time, the cell structure can be identified.

  According to the present invention, for example, an improvement in the speed and accuracy of endoscopic diagnosis can be expected by sensing a temperature change in the deep part of the body using a highly sensitive temperature-sensitive dye. In addition, by combining the present invention with a therapeutic endoscope or in combination with MRI, more accurate diagnosis and treatment than before can be achieved.

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the scope of the present invention is not limited to these examples.

<Adjustment of environmentally responsive fluorescent dye-containing copolymer particle aqueous dispersion>
15 parts by weight of organic polymer fine particles, 0.03 part by weight of tris (thenoyltrifluoroacetonato) europium (III) (Eu (III) (TTA) 3) which is a temperature sensitive fluorescent dye, fluorescein which is a non-temperature sensitive fluorescent dye 0.00001 parts by weight of isothiocyanate (FITC) and 85 parts by weight of distilled water were charged into a 100 ml autoclave, heated and stirred at 135 ° C. and 800 rpm for 30 minutes, and then cooled to room temperature while maintaining stirring. An environmentally responsive fluorescent dye-containing copolymer particle aqueous dispersion was obtained.

[Example 1]
Prepare a liquid prepared by mixing 50 parts by weight of the aqueous dispersion of environmentally responsive fluorescent dye-containing polymer particles obtained above, 25 parts by weight of 10% TMOS / methanol solution, and 3 parts by weight of 1% NH 3 solution. After coating on the tip of the optical fiber to be the probe of the apparatus, the environment-responsive fluorescent dye was immobilized on the tip of the optical fiber by heating in an oven at 100 degrees for 30 minutes.

[Example 2]
0.23 parts by weight of tris (thenoyltrifluoroacetonato) europium (III) (Eu (III) (TTA) 3), fluorescein isothiocyanate (FITC) as a non-temperature sensitive fluorescent dye 0.000077 parts by weight, 10% TMOS Prepare a mixture of 25 parts by weight of methanol / methanol solution and 3 parts by weight of 1% NH3 solution, apply it to the tip of the optical fiber that will be the probe of the diagnostic device, and then heat it in an oven at 100 degrees for 30 minutes. Fluorescent dye was immobilized on the tip of the optical fiber.

[Example 3]
Using nanoparticles encapsulating Eu-TTA and FITC, we measured the temperature of mouse acute brain sections. The leftmost is a cerebellar slice, and the other two are cerebral regions. The dark red part has a relatively high temperature and the deep green part has a low temperature. It can be seen that the tissue structure and nerve fibers can be observed by using the temperature map. By taking a relative ratio (ratio) of the fluorescence intensity of Eu-TTA relative to FITC that does not undergo a temperature-dependent fluorescence intensity change in the vicinity of 23 to 37 ° C., the temperature difference of the living body can be shown more clearly. Digitization is also possible.
Mouse sagittal sections were prepared with a thickness of 250 mm in Krebs-Ringer solution saturated with 95% O2 / 5% CO2. The microparticles used in Example 1 in which Eu-TTA and FITC were encapsulated were melted in ultrapure water, and the microparticles were applied directly to brain slices. The following is the result of observation under a microscope.

Claims (6)

A method for sensing biological tissue information using organic polymer particles encapsulating environmentally responsive dyes. A composite optical fiber containing organic polymer particles encapsulating environmentally responsive dyes. The composite optical fiber according to claim 2, wherein temperature sensing in the body is performed using the fact that an environmentally responsive dye exhibits temperature sensitivity. 4. The composite optical fiber according to claim 2, wherein the organic polymer particles are polymer particles having a 50% volume average particle diameter of 1 to 35 nm. 5. The composite optical fiber according to claim 2, wherein organic polymer particles encapsulating an environmentally responsive dye are fixed to the tip of the optical fiber using a binder. A method for immobilizing the organic polymer fine particles according to claim 5 on the tip of an optical fiber.