JP2007033090A - Optical detecting substrate and its manufacturing method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To precisely apply a sample solution on a substrate at a predetermined position so as to uniformize the size and shape of a spot in an optical detecting substrate used in a method for detecting the detection light emitted from the substance deposited on the substrate. <P>SOLUTION: Hydrophilic regions 2 are cyclically provided on the surface of the optical detecting substrate 1 and the cycle of the hydrophilic regions 2 is set to 5,000 nm or below. The optical detecting substrate 1 is manufactured by a process for forming a thin film, which changes the affinity with water by irradiation with an active energy beam, on the surface of the substrate and by a step for selectively irradiating the thin film with the active energy beam. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an optical detection substrate having a surface structure suitable for optical detection of, for example, fluorescence and a method for manufacturing the same.

In recent years, development of devices called biochips or analysis methods represented by DNA chips has become active. The biochip is used for performing analysis or the like by allowing a biopolymer such as DNA or protein to be adsorbed or supported at a specific position on a substrate and reacting with the biopolymer interacting with the biopolymer.
As a representative biochip, there is a DNA chip in which various types of DNA fragments (detection DNA) are fixed on a substrate as hundreds to tens of thousands of minute spots. The DNA chip is allowed to be evaluated (hybridization) with human or animal DNA to be evaluated (hybridization) to detect and evaluate a large number of DNA sequences at one time. Differences in gene expression due to
Hereinafter, DNA will be described as a representative example, but the same applies to RNA, protein, sugar chain, and the like.

In a DNA chip, a method for immobilizing DNA on a substrate is roughly classified into a solid phase synthesis method using photolithography and a Stanford (stamping) method in which many kinds of prepared DNAs are arranged on the substrate. .
Whichever method is used, as a detection method, a mixed DNA fragment (hereinafter referred to as sample DNA) is hybridized to DNA immobilized on a substrate (hereinafter referred to as probe DNA), and A technique is generally used in which the amount of DNA having individual sequences present in the sample DNA is estimated by reading the fluorescence from the fluorescently labeled molecules attached in advance to the sample DNA.

A typical method for immobilizing probe DNA on a substrate by the Stanford (stamping) method is to first prepare a sample solution in which probe DNA is dissolved in a buffer solution or the like at a specific position and a specific position on the substrate by a stamp. Spot the area. The fixing region of the substance fixed on the substrate in this way is hereinafter referred to as a spot. Also, this spotting operation is sometimes called a spot. Then, the probe DNA in the spot is adsorbed or covalently bonded onto the substrate (hereinafter referred to as immobilization). As a typical method for stamping a sample solution, there is a method in which a sample solution is attached to the tip of a stainless steel pin and brought into contact with the substrate surface to transfer the sample solution to the substrate surface. Thus, after the probe DNA is immobilized on the substrate and further hybridized, fluorescence detection is performed, and a detection step is performed in which the fluorescence intensity between the spots is compared.
In the detection step, generally, a method of automatically recognizing a large number of spots by an image processing program is used. Therefore, if each spot does not exist accurately at a predetermined position, a predetermined area, and a predetermined shape, spot recognition is performed. The correction operation becomes complicated, and a sufficiently accurate result cannot be obtained.
That is, in the DNA chip, the accuracy of the spot is an important point for obtaining good analysis results.

For example, Patent Document 1 described below describes a method of obtaining a biochip by forming a site having a hydrophilic surface at a high density on a substrate and attaching a biological substance to the site.
JP2003-121442A

  However, in the method described in Patent Document 1, since the spot shape and arrangement are the same as the site shape and arrangement, a substrate on which a desired site pattern is formed is prepared each time the spot shape or arrangement is changed. There was a need to do.

  The present invention is for optical detection capable of spotting a sample solution at a predetermined position on a substrate with high accuracy so that the spot size and shape are uniform, and easily adapting to changes in spot shape and arrangement. A substrate and a manufacturing method thereof are provided.

In order to solve the above problems, an optical detection substrate of the present invention is a substrate used in a method for detecting detection light emitted from a substance fixed on a substrate, and has a hydrophilic region on the surface of the substrate. It is provided periodically, and the period of the hydrophilic region is 5000 nm or less and is smaller than the diameter of the fixing region of the substance.
The method for producing an optical detection substrate of the present invention comprises a step of forming a thin film whose affinity with water is changed by irradiation of active energy rays on the surface of the substrate, and selective selection of active energy rays with respect to the thin film. It has the process of irradiating to.

  According to the present invention, the sample solution can be spotted at a predetermined position on the substrate with high accuracy so that the size and shape of the spot are uniform, and can be easily adapted to changes in the shape and arrangement of the spot. A detection substrate is obtained.

FIG. 1 is a plan view showing a first embodiment of an optical detection substrate (hereinafter sometimes simply referred to as a substrate) of the present invention. The drawing schematically shows an enlarged part of the surface of the substrate 1. Two directions perpendicular to each other in a plane perpendicular to the thickness direction of the substrate 1 are defined as an X direction and a Y direction, respectively.
In the substrate 1 of this embodiment, a rectangular hydrophilic region 2 is periodically formed on the surface along two repeating directions of the X direction and the Y direction. The periphery of the hydrophilic region 2 is surrounded by the hydrophobic region 3 and has a sea-island structure.
In the present embodiment, the shape and size of the hydrophilic region 2 are uniform. That is, the width Wx of the hydrophilic region 2 in the X direction is constant, and the width Wy of the hydrophilic region 2 in the Y direction is also constant.
The interval Sx between the hydrophilic regions 2 in the X direction is constant, and the interval Sy between the hydrophilic regions 2 in the Y direction is also constant.

The hydrophilic area | region 2 in this invention means the area | region whose surface contact angle with respect to water is 80 degrees or less. The contact angle with respect to water in the hydrophilic region 2 is preferably 70 degrees or less, and more preferably 60 degrees or less.
The hydrophobic region 3 refers to a region having a surface contact angle with water exceeding 80 degrees. The contact angle with respect to water in the hydrophobic region 3 is preferably 100 degrees or more, and more preferably 110 degrees or more. In this specification, the case where the contact angle with respect to water is 100 degrees or more may be said to have “water repellency”.
In obtaining the effect of the present invention, the difference in contact angle with respect to water between the hydrophilic region 2 and another region in contact with the hydrophilic region 2 (the hydrophobic region 3 in this embodiment) is 30 degrees or more. Is preferable, and 50 degrees or more is more preferable.

The repeating direction of the hydrophilic region 2 in the present invention is determined as follows. First, paying attention to an arbitrary first hydrophilic region 2, the second hydrophilic region 2 having the shortest distance among the adjacent hydrophilic regions 2 is selected. A first repetition of a direction from an arbitrary point A1 in the first hydrophilic region 2 to a corresponding point A2 in the second hydrophilic region 2 (a point at the same position overlapping by parallel movement in one direction) The direction. A distance P1 from the point A1 to the point A2 is the first period.
Next, of the hydrophilic regions 2 adjacent to the first hydrophilic region 2, the third hydrophilic region 2 having the shortest distance is selected except for those in the first repeating direction. A direction from an arbitrary point A1 in the first hydrophilic region 2 to a corresponding point A3 in the third hydrophilic region 2 is defined as a second repeating direction. A distance P2 from the point A1 to the point A3 is the second period.
There are two cycles of the hydrophilic region 2 in the present embodiment: a first cycle in the X direction (P1 = (Wx + Sx) value) and a second cycle in the Y direction (P2 = (Wy + Sy) value). To do.

In the present invention, the period of the hydrophilic region 2 is 5000 nm or less, preferably 1000 nm or less.
When there are a plurality of cycles of the hydrophilic region and the plurality of cycles are different from each other, the value of at least one cycle is 5000 nm or less, preferably 1000 nm or less. More preferably, the entire period is within the above range.
By setting the period of the hydrophilic region 2 within the above range, it is possible to suppress the occurrence of gathering at the outer edge of the spot shape. This is preferable for obtaining a spot shape that is circular, preferably a perfect circle. Further, if the period of the hydrophilic region 2 is within the above range, the pattern formed on the surface of the substrate 1 becomes a finer pattern than the general reading resolution in the optical detection device. An increase in background noise in optical detection can be prevented.

  The lower limit value of the period of the hydrophilic region 2 is not particularly limited. However, in order to sufficiently obtain the effect of the present invention by providing the distribution of the hydrophilic region 2 and the hydrophobic region 3 on the surface of the substrate 1, hydrophilicity is required. It is preferable that the period of the region 2 is larger than the molecule contributing to the expression of hydrophilicity and / or water repellency. From this point, the lower limit of the period of the hydrophilic region 2 is preferably 10 nm or more, more preferably 30 nm or more, and further preferably 100 nm or more.

The widths of the hydrophilic regions 2 in different repeating directions, that is, in the present embodiment, the width Wx in the X direction and the width Wy in the Y direction may be the same or different. Further, the spacing between the hydrophilic regions 2 in different repeating directions (the width of the hydrophobic region 3), that is, the spacing Sx in the X direction and the spacing Sy in the Y direction may be the same or different in this embodiment.
In order to make the spot shape of the sample solution spotted on the substrate into a circular shape, preferably a perfect circular shape, hydrophilic regions 2 are periodically provided in two different repeating directions, It is preferable that the period of the hydrophilic region 2 in the repeating direction and the period in the second repeating direction are the same (P1 = P2 in the present embodiment). In this specification, this state is referred to as the two-dimensional isotropic property of the periodic structure of the hydrophilic region 2.
In the periodic structure having the two-dimensional isotropic property, the width of the hydrophilic region 2 in the first repeating direction is the same as the width of the hydrophilic region 2 in the second repeating direction (Wx = Wy in this embodiment). In addition, it is spotted on the substrate that the interval between the hydrophilic regions 2 in the first repeating direction and the interval between the hydrophilic regions 2 in the second repeating direction are the same (Sx = Sy in this embodiment). It is more preferable for the spot shape of the sample solution to be a circle, preferably a perfect circle.

In the example of FIG. 1, the planar shape of the hydrophilic region 2 is a rectangle, but the shape is not limited to this and can be changed as appropriate. 2 and 3 show a modification. Arrows (1) and (2) in the figure indicate two different repeating directions perpendicular to the thickness direction of the substrate, arrow (1) is the first repeating direction, and arrow (2) is Each of the second repetition directions is shown.
The example of FIG. 2 is an example in which the planar shape of the hydrophilic region 2 is circular. That is, the circular hydrophilic region 2 is periodically formed with a constant period along the first repeating direction (1) and the second repeating direction (2). The periphery of the hydrophilic region 2 is surrounded by the hydrophobic region 3 and has a sea-island structure. The shape and size of the hydrophilic region 2 are uniform.
In FIG. 2, the period in the first repetition direction is indicated by P1, and the period in the second repetition direction is indicated by P2.
1 and 2, in a sea-island structure in which the hydrophilic region 2 is surrounded by the hydrophobic region 3, the first repeating direction (X direction, (1) direction) and the second repeating direction (Y direction). , (2) direction), the width (Wx, Wy, W1, W2) of the hydrophilic region 2 is preferably 50 nm or more, and more preferably 100 nm or more, when the sample solution is brought into close contact with the substrate. The width (Sx, Sy, S1, S2) of the hydrophobic region 3 is preferably 10 nm or more, more preferably 30 nm or more, and further preferably 50 nm or more, in order to prevent unexpected wetting and spreading of the sample solution.

In the example of FIG. 3, the planar shape of the hydrophilic region 2 is rectangular, the hydrophilic regions 2 are in contact with each other at points, and the four sides of the hydrophilic region 2 are in contact with the rectangular hydrophobic region 3, respectively. It is. The shape and size of the hydrophilic region 2 and the hydrophobic region 3 are uniform.
In FIG. 3, the period in the first repetition direction is indicated by P1, and the period in the second direction is indicated by P2.

The sea-island structure shown in FIGS. 1 and 2 in which the hydrophobic region 3 is present all around the hydrophilic region 2 has a circular shape, preferably a perfect circular shape, of the sample solution spotted on the substrate. More preferable.
On the other hand, the structure shown in FIG. 3 where the hydrophilic regions 2 are in contact with each other at a point is more preferable in that it is easy to increase the ratio of the area of the hydrophilic region 2 to the hydrophobic region 3.
As the repetitive pattern, a simple square pattern in which the first repetitive direction and the second repetitive direction are orthogonal as in the examples of FIGS. 1 and 3, and the first repetitive direction and the second as in the example of FIG. A hexagonal filling pattern in which the angle formed by the repeating direction is 60 ° is preferable, but other patterns can be employed.

The material constituting the substrate 1 is not particularly limited as long as it can form a pattern composed of the hydrophilic region 2 and the hydrophobic region 3 on the surface. The substrate 1 may be transparent or opaque in a range of 450 nm to 700 nm, which is a fluorescent wavelength region of a fluorescent label typically used for a biochip.
Specific examples of the transparent material include glass materials such as quartz glass, soda lime glass, and borosilicate glass; a fluororesin; and a silicon-containing resin.
Examples of opaque materials include metals such as stainless steel.
Among these, borosilicate glass or soda lime glass is more preferable particularly in terms of low fluorescence background, flatness, and availability.

  As a method of manufacturing the optical detection substrate of the present invention, a step of forming a thin film whose affinity with water is changed by irradiation of active energy rays on the surface of the substrate 1, and an active energy ray is applied to the thin film. A method having a step of selectively irradiating is preferable.

  Specifically, a water-repellent thin film is formed on the surface of the substrate 1 using a water-repellent composition containing a fluorine-containing compound represented by the following formula 1 (hereinafter also referred to as compound 1) and an organic solvent. Then, by irradiating the water-repellent thin film with ultraviolet rays, the irradiated portion of the fluorine-containing compound (compound 1) is decomposed, and the decomposed product is removed to form a pattern comprising a hydrophilic region and a water-repellent region. The forming method is preferred.

[R ¹ , R ² , R ³ , and R ^{4 in} Formula 1 are each independently a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, or a monovalent organic group, and R ¹ , R ² , R ³ , R ^At least one of ⁴ is an organic group having a fluorine atom. R ⁵ is a hydrogen atom or a monovalent organic group, R ⁶ is a monovalent organic group having a hydrolyzable group, and A is an oxygen atom or —NH—. ]

In the compound 1, at least one of R ¹ , R ² , R ³ , and R ⁴ is a monovalent organic group containing a fluorine atom, and the monovalent organic group containing a fluorine atom includes R ^F —B A group represented by-(R ^F is a polyfluoroalkyl group (hereinafter referred to as R ^F group) and B represents a divalent group or a covalent bond not containing a fluorine atom) is preferable. The ^RF group refers to a group in which two or more hydrogen atoms of an alkyl group are substituted with fluorine atoms. R ^F number of carbon atoms of the group is preferably 1 to 20, more preferably 4 to 16, 6 to 12 being most preferred. When the number of carbon atoms is within this range, the formed thin film is excellent in water repellency.
The ^RF group may have a linear structure, a branched structure, or a ring structure, and a linear structure is preferable. In the case of a branched structure, the number of carbon atoms in the branched portion is preferably 1 to 4. Further, the R ^F group may have an unsaturated bond. The ^RF group may have a halogen atom other than the fluorine atom, and the chlorine atom is preferred as the other halogen atom. Also, carbon in the R ^F group - is between carbon bond, an etheric oxygen atom or thioetheric sulfur atom may be inserted.
The number of fluorine atoms in R ^F group is represented by a (R ^F number of fluorine atoms in the group) / (R ^F number of hydrogen atoms in the alkyl group corresponding to the same number of carbon atoms and groups) × 100 (%) 60% or more is preferable, and 80% or more is particularly preferable.

Specific examples of the R ^F group are listed below, but are not limited thereto.
_{CF 3 -, F (CF 2} ) 2 -, F (CF 2) 3 -, F (CF 2) 4 -, F (CF 2) 5 -, F (CF 2) 6 -, F (CF 2) 7 _{-, F (CF 2) 8} -, F (CF 2) 9 -, F (CF 2) 10 -, F (CF 2) 11 -, F (CF 2) 12 -, F (CF 2) 13 -, _{F (CF 2) 14 -,} F (CF 2) 15 -, F (CF 2) 16 -, (CF 3) 2 CF -, (CF 3) 2 CFCF 2 -, (CF 3) 2 CFCF 2 CF 2 _{-, (CF 3) 2 CFCF} 2 CF 2 CF 2 CF 2 -, (CF 3) 2 CFCF (CF 3) -, (CF 3) 2 CFCF (CF 3) CF 2 -, (CF 3) 2 CFCF 2 _{CF (CF 3) -, (} CF 3) 2 CFCF 2 CF (CF 3) CF 2 CF 2 -, _{F 3 CF 2 CF (CF 3} ) -, CF 3 CF 2 CF (CF 3) CF 2 CF 2 -, CF 3 CF 2 CF (CF 2 CF 3) -, CF 3 CF 2 CF (CF 2 CF 3) _{CF 2 CF 2 -, (CF} 3) 3 C -, (CF 3) 3 CCF 2 -, (CF 3) 3 C (CF 2 CF 3) -, (CF 2 CF 3) C (CF 3) 2 - _{, (CF 2 CF 3) 2} CCF 3 -.

_{CF 2 = CF-, CF 3 CF} = CF-, CF 3 CF 2 CF = CF-, CF 3 CF 2 CF 2 CF = CF-, CF 3 CF 2 CF 2 CF 2 CF = CF-, CF 3 CF 2 _{CF 2 CF 2 CF 2 CF =} CF-, CF 3 CF 2 CF 2 CF 2 CF 2 CF 2 CF = CF-, CF 3 C (CF 3) = CF-, CF 3 C (CF 3) = C (CF ₃ )-.
HCF _2- , HCF ₂ CF _2- , HCF ₂ CF ₂ CF _2- , HCF ₂ CF ₂ CF ₂ CF _2- , HCF ₂ CF ₂ CF ₂ CF ₂ CF ₂ CF _2- , HCF ₂ CF ₂ CF ₂ CF _{2 CF 2 CF 2 CF 2 CF 2} -.

_{ClCF 2 -, ClCF 2 CF 2} -, ClCF 2 CF 2 CF 2 -, ClCF 2 CF 2 CF 2 CF 2 -, ClCF 2 CF 2 CF 2 CF 2 CF 2 -, ClCF 2 CF 2 CF 2 CF 2 CF 2 _{CF 2 -, ClCF 2 CF 2} CF 2 CF 2 CF 2 CF 2 CF 2 -, ClCF 2 CF 2 CF 2 CF 2 CF 2 CF 2 CF 2 CF 2 -.

_{CF 3 OCF 2 -, CF 3} CF 2 OCF 2 -, CF 3 OCF 2 CF 2 -, CF 3 CF 2 OCF 2 CF 2 -, CF 3 CF 2 OCF 2 CF 2 OCF 2 -, CF 3 CF 2 OCF 2 _{CF 2 OCF 2 CF 2 -,} CF 3 CF 2 OCF 2 CF 2 OCF 2 CF 2 OCF 2 -, CF 3 CF 2 OCF 2 CF 2 OCF 2 CF 2 OCF 2 CF 2 -, CF 3 CF 2 OCF 2 CF 2 OCF ₂ CF ₂ OCF ₂ CF ₂ OCF _2- , CF ₃ CF ₂ OCF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF _2- , CF ₃ CF ₂ CF ₂ OCF _2- , CF ₃ CF ₂ CF _{2 CF 2 OCF 2 -, CF 3} CF 2 CF 2 CF 2 CF 2 CF 2 OCF 2 -, CF 3 CF 2 CF 2 OCF _{CF 2 -, CF 3 CF 2} CF 2 CF 2 OCF 2 CF 2 -, CF 3 CF 2 CF 2 CF 2 CF 2 CF 2 OCF 2 CF 2 -, CF 3 CF 2 CF 2 CF 2 OCF 2 CF 2 OCF 2 _{-, CF 3 CF 2 CF 2} CF 2 OCF 2 CF 2 OCF 2 CF 2 -, CF 3 CF 2 CF 2 CF 2 CF 2 CF 2 OCF 2 CF 2 OCF 2 -. CF ₃ CF ₂ CF ₂ CF ₂ CF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF _2- , CF ₃ CF ₂ CF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF ₂ OCF _2- , CF ₃ CF ₂ CF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF _2- , CF ₃ CF ₂ CF ₂ CF ₂ CF ₂ CF ₂ OCF ₂ CF ₂ OCF ₂ CF ₂ OCF _2- , CF ₃ CF ₂ CF ₂ CF ₂ CF ₂ CF _{2 OCF 2 CF 2 OCF 2 CF 2} OCF 2 CF 2 -, CF 3 CF 2 CF 2 OCF (CF 3) -, CF 3 CF 2 CF 2 OCF (CF 3) CF 2 -, CF 3 CF 2 CF 2 OCF ( _{CF 3) CF 2 OCF 2 CF} 2 -, CF 3 CF 2 CF 2 OCF (CF 3) CF 2 OCF (CF 3) -, CF 3 _{F 2 CF 2 OCF (CF 3} ) CF 2 OCF (CF 3) CF 2 -.

_{CF 3 CF 2 OCF = CF-,} CF 3 CF 2 OCF 2 CF 2 OCF = CF-, CF 3 CF 2 CF 2 OCF = CF-, CF 3 CF 2 CF 2 CF 2 OCF = CF-, CF 3 CF 2 _{CF 2 CF 2 OCF 2 CF 2} OCF = CF-, CF 3 CF 2 CF 2 CF 2 CF 2 CF 2 OCF = CF-, CF 3 CF 2 CF 2 CF 2 CF 2 CF 2 OCF 2 CF 2 OCF = CF- , CF ₃ CF ₂ CF ₂ OCF (CF ₃ ) CF ₂ OCF═CF—.

B in R ^F —B— is a divalent group or a covalent bond. The divalent group may contain an oxygen atom, a nitrogen atom, a sulfur atom, a phosphorus atom, a chlorine atom, a bromine atom, or an iodine atom, and may be a saturated group, an unsaturated group, or a linear structure. A branched structure or a ring structure may be used. The site | part couple | bonded with a benzene ring can take a resonance structure, and an oxygen atom, a nitrogen atom, a double bond, and a carbonyl group are preferable as a resonance site | part.

Specific examples of R ^F —B— are listed below, but are not limited thereto.
R ^F — (CH ₂ ) _q O— (q is an integer of 1 to 10), R ^F —NH—, R ^F —CH ₂ NH—, R ^F —CH ₂ CH ₂ NH—, R ^F —CH ₂ CH ₂ CH ₂ NH—, R ^F — (CH ₂ ) ₄ NH—, R ^F —N (CH ₃ ) —, R ^F —CH ₂ N (CH ₃ ) —, R ^F —CH ₂ CH ₂ N (CH ₃ )-, R ^F —CH ₂ CH ₂ CH ₂ N (CH ₃ ) —, R ^F —CH ₂ CH ₂ CH ₂ CH ₂ N (CH ₃ ) —.

R ^F —OC (═O) —, R ^F —CH ₂ OC (═O) —, R ^F —CH ₂ CH ₂ OC (═O) —, R ^F —CH ₂ CH ₂ CH ₂ OC (═O) −, R ^F —CH ₂ CH ₂ CH ₂ CH ₂ OC (═O) —, R ^F —C (═O) —, R ^F —CH ₂ C (═O) —, R ^F —CH ₂ CH ₂ C (═O) —, R ^F —CH ₂ CH ₂ CH ₂ C (═O) —, R ^F —CH ₂ CH ₂ CH ₂ CH ₂ C (═O) —, R ^F —C (═O) O— , R ^F —CH ₂ C (═O) O—, R ^F —CH ₂ CH ₂ C (═O) O—, R ^F —CH ₂ CH ₂ CH ₂ C (═O) O—, R ^F —CH _{2 CH 2 CH 2 CH 2 C} (= O) O-.

R ^F —C (═O) NH—, R ^F —CH ₂ C (═O) NH—, R ^F —CH ₂ CH ₂ C (═O) NH—, R ^F —CH ₂ CH ₂ CH ₂ C ( ═O) NH—, R ^F —CH ₂ CH ₂ CH ₂ CH ₂ C (═O) NH—, R ^F —CH═CH—, R ^F —CH ₂ CH═CH—, R ^F —CH ₂ CH ₂ CH = CH-.
Among these, R ^F — (CH ₂ ) _q O— (q is an integer of 1 to 10) is preferable, R ^F — (CH ₂ ) ₃ O— is more preferable, and C ₈ F ₁₇ (CH ₂ ) ₃ O -Is most preferred.

In R ¹ , R ² , R ³ , and R ⁴ in Compound ¹ , the monovalent organic group other than the monovalent organic group having a fluorine atom is preferably a monovalent organic group having an oxygen atom, and more preferably an alkoxy group. , Nitrogen atom, sulfur atom, phosphorus atom, chlorine atom, bromine atom, iodine atom. Further, it may be a saturated group or an unsaturated group, and may have any of a straight chain structure, a branched structure, and a ring structure. The monovalent organic group is preferably a group capable of having a resonance structure with a heteroatom or a benzene ring because it excels in light absorption characteristics of the benzene ring. The number of monovalent organic groups in R ¹ , R ² , R ³ and R ⁴ is preferably 1 or more.
It is not particularly restricted but includes specific _{examples, -OCH 3, -OCH 2 CH 3} , -OCH 2 CH 2 CH 3, -OCH (CH 3) 2, -NH (CH 3), - N (CH 3) 2, _{-CN, -C (= O) OH} , -C (= O) OCH 3, -OC (= O) CH 3, -NHC (= O) CH 3, -N (CH 3) C (= O) CH ₃ is preferred.

One or more of R ¹ , R ² , R ³ , and R ⁴ are preferably hydrogen atoms, and more preferably 2 or more are hydrogen atoms.

In the compound 1, R ⁵ is a hydrogen atom or a monovalent organic group, preferably a hydrogen atom. When it is a monovalent organic group, a lower alkyl group is preferable, and a methyl group or an ethyl group is more preferable.
In compound 1, A is more preferably an oxygen atom. When A is an oxygen atom, the thin film after decomposition by ultraviolet rays is excellent in hydrophilicity.

In Compound 1, R ⁶ is a monovalent organic group having a hydrolyzable group, and may contain a silicon atom, an oxygen atom, a nitrogen atom, a sulfur atom, a phosphorus atom, a chlorine atom, a bromine atom, or an iodine atom, A saturated group or an unsaturated group may be used, and any of a linear structure, a branched structure, and a ring structure may be sufficient.
The R ^6, preferably a group having a silicon _{^{atom, -R 7 -Si (R 8)}} 3-m X m or ^{-R 7} -SH ^(R 7 is a divalent organic group, ^{R 8} is a monovalent organic is a group, X is a hydrolyzable group, m is more preferably a group represented by an integer of 1 to ^3.), and most preferably _{^{-R 7 -Si (R 8) 3}} -m X m . R ⁷ is preferably an alkylene group having 1 to 10 carbon atoms, and —C (═O) — or —NH— may be inserted between carbon-carbon bonds of the alkylene group. R ⁸ is preferably a lower alkyl group. X is preferably an alkoxy group, an acyloxy group, a ketoxime group, an alkenyloxy group, an amino group, an aminoxy group, an amide group, an isocyanate group or a halogen atom, more preferably a halogen atom (especially a chlorine atom) or an alkoxy group, and an alkoxy group Is most preferred. As an alkoxy group, a C1-C3 alkoxy group is preferable, and a methoxy group or an ethoxy group is more preferable. As m, 2 or 3 is preferable and 3 is more preferable. The more m, the better the adhesion with the substrate.

  As specific examples of compound 1, the following compounds are preferably exemplified. Hereinafter, the methyl group may be expressed as Me and the ethyl group as Et.

Compound 1 contains a monovalent organic group having a fluorine atom, and a thin film formed using Compound 1 is excellent in water repellency. The compound 1 is decomposed by ultraviolet rays, and the contact angle of the thin film with water is reduced by ultraviolet irradiation. This is considered to be because when Compound 1 is irradiated with ultraviolet rays, Compound 1 is decomposed into -A-R ⁶ and other moieties, and -A-R ⁶ is changed to a hydroxyl group or an amino group. .

The water repellent composition used to form a water repellent thin film on a substrate contains Compound 1 and an organic solvent. The water repellent composition may contain one type of compound 1 or two or more types.
The organic solvent is not particularly limited, and alcohols, ketones, aromatic hydrocarbons, and paraffinic hydrocarbons are preferable, and lower alcohols such as ethyl alcohol and 2-propyl alcohol or paraffinic hydrocarbons are more preferable. preferable. One organic solvent may be used, or two or more organic solvents may be mixed and used to adjust polarity, evaporation rate, and the like.
The content of the organic solvent in the water repellent composition is preferably 0.01 to 1000 parts by mass, more preferably 0.1 to 100 parts by mass with respect to 1 part by mass of the compound 1. Within this range, a uniform thin film is formed and excellent water repellency is exhibited.
The water-repellent composition may be further diluted with an organic solvent as necessary in consideration of workability, the thickness of the thin film to be formed, and the like.

The water repellent composition may contain components other than Compound 1 and the organic solvent. The other component is preferably an acid, preferably an inorganic acid such as sulfuric acid, hydrochloric acid, nitric acid or phosphoric acid, or an organic acid such as acetic acid, formic acid, p-toluenesulfonic acid or methanesulfonic acid. When R ⁶ has a hydrolyzable group bonded to a silicon atom, by adding an acid to the water-repellent composition, the hydrolyzable group is converted to a silanol group (Si—OH), Si-O-Si is formed by dehydration condensation reaction. At this time, if a Si—OH group is present on the substrate, it reacts with it to improve the adhesion to the substrate.
The content of the acid in the water-repellent composition is preferably 0.0001 to 0.1 parts by weight, more preferably 0.001 to 0.01, with respect to 1 part by weight of the compound 1. When it is within this range, the hydrolysis effect is sufficiently exhibited, and the composition is excellent in stability.

In order to form the hydrophilic region 2 on the substrate 1 using the water-repellent composition containing the compound 1, first, the water-repellent composition is applied onto the substrate 1 to form a water-repellent thin film.
It is preferable that the substrate 1 is appropriately pretreated in advance, such as a cleaning process using an abrasive.
Examples of the method for applying the water-repellent composition include various known methods such as brush coating, flow coating, spin coating, dip coating, squeegee coating, spray coating, and hand coating. The water repellent composition is applied onto the substrate 1 and then dried in the air or in a nitrogen stream. The drying is preferably performed at room temperature. When drying by heating, the heating temperature and time are set in consideration of the heat resistance of the substrate 1.
The thickness of the water repellent thin film is not particularly limited, but is preferably 100 nm or less, more preferably 20 nm or less, and most preferably 5 nm or less. The thickness of the water repellent hydrophilic thin film is preferably about the same as the thickness (about 1 to 3 nm) of a self-assembled monolayer (hereinafter referred to as SAM; SAM = Self-Assembled Monolayer). When the film thickness of the water-repellent thin film is in the above range, in the subsequent ultraviolet irradiation step, ultraviolet rays are efficiently irradiated to the compound, so that the decomposition reaction proceeds efficiently.

Next, the water repellent thin film is selectively irradiated with ultraviolet rays to decompose the irradiated portion of the compound 1.
The wavelength of the irradiated ultraviolet light is preferably 200 nm or more, more preferably 250 nm or more, and most preferably 300 nm or more. Ultraviolet rays having a wavelength of less than 200 nm may decompose the material constituting the substrate 1 itself. As the ultraviolet light source, a low-pressure mercury lamp, a high-pressure mercury lamp, an ultrahigh-pressure mercury lamp, a xenon lamp, a sodium lamp, a gas laser such as nitrogen, a liquid laser of an organic dye solution, a solid containing rare earth ions in an inorganic single crystal A laser etc. are mentioned preferably. In addition, as a light source other than a laser capable of obtaining monochromatic light, a broadband line spectrum or continuous spectrum can be used. In this case, an ultraviolet filter having a necessary wavelength is used by using an optical filter such as a bandpass filter or a cutoff filter. It is preferable to take out and use. In particular, a high-pressure mercury lamp or an ultrahigh-pressure mercury lamp is preferable as the light source because a large area can be irradiated at one time.
Examples of the method of selectively irradiating ultraviolet rays include a method of irradiating ultraviolet rays through a photomask having a predetermined pattern and a method using laser light. Since a large area can be irradiated at a time, a method of irradiating ultraviolet rays through a photomask is preferable.

After the irradiation with ultraviolet rays, it is preferable to wash the substrate 1 in order to remove decomposition products of the compound 1. As the cleaning liquid used for the cleaning, for example, acetone and dichloropentafluoropropane (R-225) are preferably used in this order.
Of the water-repellent thin film formed on the substrate 1, the region where the compound 1 is decomposed by being selectively irradiated with ultraviolet light is more water-soluble than the water-repellent thin film (hydrophobic region 3) which is not irradiated with ultraviolet light. The contact angle with respect to is lowered, and the hydrophilic region 2 is obtained. Thus, the optical detection substrate of the present invention is obtained.
The water contact angle of the water-repellent thin film (hydrophobic region 3) formed using the water-repellent composition containing Compound 1 is preferably 100 degrees or more, and more preferably 110 degrees or more. Further, the contact angle with water in the region (hydrophilic region 2) where the decomposition of the compound 1 is caused by ultraviolet irradiation is preferably 80 degrees or less, more preferably 70 degrees or less, and most preferably 60 degrees or less.
Further, the difference in the contact angle of the water-repellent thin film with respect to water before and after being decomposed by ultraviolet irradiation (the difference in contact angle between the hydrophilic region 2 and the hydrophobic region 3) is preferably 30 ° or more, and more preferably 50 ° or more.

  Since a hydroxyl group or an amino group is present on the surface of the hydrophilic region 2 formed by the above method, it is easy to adsorb and fix a biopolymer or the like. Further, the substrate 1 and the biopolymer (for example, a probe DNA in a DNA chip) are reacted with an organic substance having reactivity with a hydroxyl group or an amino group and having a high affinity with the biopolymer. ) And / or binding specificity is improved, and fixing of the biopolymer to the substrate 1 is promoted.

In the optical detection substrate of the present invention, the surface of the hydrophilic region 2 may be subjected to a surface treatment that promotes fixing of a substance fixed on the substrate.
That is, after forming a thin film on the entire surface of the substrate 1 using the water repellent composition and forming the hydrophilic region 2 by partially irradiating with ultraviolet rays, preferably after removing decomposition products after irradiating with ultraviolet rays. The hydrophilic region 2 may be subjected to a surface treatment.
The surface treatment agent used for the surface treatment preferably has a function of accelerating the fixing of a substance (for example, probe DNA immobilized on the substrate) to the substrate 1 to the substrate 1, and the kind of the substance It is appropriately selected depending on.

The optical detection substrate of the present invention is used in a method for detecting detection light emitted from a substance fixed on the substrate 1. As detection, qualitative analysis, quantitative analysis, and the like can be performed.
In the present specification, a substance to be fixed on the substrate 1 (hereinafter sometimes referred to as a substance to be fixed).
) Refers to a substance fixed on the substrate 1 at the time of detection. In addition to the substance immobilized on the substrate 1 in advance, such as probe DNA in a DNA chip, It is a concept that includes a substance to be detected such as a sample DAN to be hybridized, and a substance in which the substance to be detected and another substance are combined.

The substance (fixed substance) to be fixed on the substrate is not particularly limited, and examples thereof include cells, DNA, RNA, proteins, organic compounds, and inorganic compounds.
Specific examples of cells include human cells, cells derived from animals and plants, bacteria and archaea such as viruses and molds.
Specific examples of the protein include water-soluble protein, keratin, collagen, fibroin, sericin, glycoprotein, membrane protein such as bacteriorhodopsin, coat protein, albumin, and serum.
Specific examples of DNA and RNA include cDNA extracted from biological material and amplified, and artificially synthesized oligo DNA.
Specific examples of the organic compound include silicon-containing compounds containing amino groups, silicon-containing compounds containing alkoxy groups, silicon-containing compounds such as silicon-containing compounds containing fluorine; sulfonic acid groups, carboxylic acid groups, hydroxyl groups, ester groups, and Fluorine compounds containing alkyl groups, etc .; fluorescent dyes; fullerenes and fullerene derivatives; organometallic compounds containing transition metals (especially organometallic compounds containing rare earth elements); macrocyclic compounds such as crown ethers, phenanthrolines, and porphyrins; Can be mentioned.
Specific examples of the inorganic compound include metal oxide fine particles such as titanium oxide and silicon dioxide; metal nitride such as titanium nitride; metal fine particles such as platinum, gold, silver, copper, and germanium; and carbon compounds.
The substance fixed on the substrate may be one kind or two or more kinds. The optical detection substrate of the present invention is suitably used as, for example, a DNA chip, a protein chip, a cell chip and the like.

The substrate of the present invention is suitable for a method of spotting droplets of a sample solution containing a material to be fixed on the substrate.
The sample solution may be any of a solution, a colloid solution, a suspension, a dispersion, and the like. As the solvent and the dispersion medium, water or an aqueous solution such as a buffer solution is used.
As a specific spotting method, for example, a method is used in which a droplet of a sample solution is attached to the tip of a stainless steel pin and the droplet is moved to the substrate surface by contacting the droplet with the substrate surface. be able to.
Alternatively, the sample solution may be spotted at a predetermined position on the substrate by a method of discharging a certain amount of droplets such as an ink jet.

The spot shape is preferably circular because image processing in optical detection is easy, and the closer to a perfect circle, the more preferable.
In addition, the size of the spot when viewed from above, that is, the size of the fixing region, is sufficient to secure a sufficient number of spots on the substrate while ensuring a sufficient amount of the material to be fixed to one spot. In view of securing the thickness, the largest diameter (major axis) is preferably in the range of 50 to 500 μm, and more preferably in the range of 80 to 200 μm.

If necessary, the sample solution (droplet) spotted on the substrate may be dried to distill off the solvent or the like in the droplet. That is, the substance fixed on the substrate may be a dried product from which the solvent is removed.
In addition, the sample solution spotted on the substrate 1 is also subjected to a treatment called deactivating a chemical active point that is no longer necessary on the substrate, which is called blocking, with respect to a substance fixed on the substrate. You may give it.

  The method for detecting the detection light emitted from the substance fixed on the substrate is not particularly limited, and various known methods can be used. For example, a method of scanning the substrate by moving the substrate in a plane direction in a measurement apparatus equipped with an optical system of a confocal microscope, or a part or all of the substrate plane is taken into the measurement apparatus at once by an image sensor such as a CCD. Method, etc.

Since the optical detection substrate of the present invention is periodically provided with fine hydrophilic regions on the surface, the sample solution is placed at a predetermined position on the substrate so that the spot size and shape are uniform. Can spot well.
For example, the preferred spot size when spotting the sample solution on the substrate is in the above range, whereas the period of the hydrophilic region provided on the substrate of the present invention is sufficiently (1 Smaller than digits) For this reason, the sample solution spotted on the substrate spreads so as to cover the plurality of hydrophilic regions 2 and the adjacent hydrophobic regions 3 due to surface tension. At this time, since it is easy to spread on the hydrophilic region 2 and difficult to spread on the hydrophobic region 3, the spread of the sample solution is regulated thereby. Therefore, when the composition of the sample solution is the same, a spot having a uniform size and shape can be obtained, and the size and shape can be determined by the end face radius of the stainless steel pin, for example.
The period of the hydrophilic region 2 provided on the substrate 1 is smaller than the size of the spot to be formed (the diameter of the fixing region), for example, 1/10 or less, preferably 1/50 or less. Since it is small, the spot is formed by a method in which a sample solution forming one spot is spotted so as to cover a plurality of hydrophilic regions 2 at once. Therefore, by using the same substrate and appropriately selecting the position and number of hydrophilic regions 2 included in one spot, the spot shape and arrangement can be freely set. Can easily handle changes.
In addition, since the periodic pattern of the hydrophilic region 2 is a finer pattern than the general reading resolution in an optical detection device, detection is performed with improved spot accuracy without increasing background noise in optical detection. Accuracy can be improved.

By the way, when the entire surface of the substrate is hydrophobic, the spotted sample solution is bounced and approaches a spherical shape (the contact angle with the substrate surface increases), so that the contact area with the substrate decreases, and the probe DNA It becomes difficult to fix the material to be fixed on the substrate surface. Further, when the sample solution is bounced, the spot position is likely to fluctuate and / or the spot shape is easily deformed.
On the other hand, when the entire surface of the substrate is hydrophilic, the sample solution wets and spreads more than necessary on the substrate, resulting in poor spot position and / or shape accuracy. In addition, since the specific surface area of the sample solution increases, the sample solution may be quickly dried. As a result, immobilization of the material to be fixed may be incomplete, or the distribution of the material to be fixed may be uneven in the spot. . Furthermore, when the spot positions of different sample solutions are close, the sample solutions may come into contact with each other and mix.
On the other hand, since the substrate of the present invention is periodically provided with fine hydrophilic regions on the surface, the spotted sample solution is not repelled and becomes almost spherical, and the amount of the sample solution Depending on the, it spreads with good reproducibility. In addition, it is possible to secure a moderately large contact area between the sample solution and the substrate, and it does not wet and spread more than necessary, so that the material to be fixed is well fixed on the substrate, and the material to be fixed in the spot is fixed. The material distribution tends to be uniform. Also, unexpected contact between adjacent spots is prevented.

In addition, when the hydrophilicity of the entire surface of the substrate is uniform, the chemical affinity between the substrate and the material to be fixed and the physical properties can be improved by selectively performing a surface treatment that promotes fixing of the material to be fixed. In order to achieve both the easy spot and the ease, it is very difficult to control by strict production management. In addition, the hydrophilicity imparted to the substrate surface by the surface treatment is also reduced by a very small amount of contaminants adhering from the air. In order to maintain the properties at the same time, the packaging of the substrate must be strict, and the expiration date of the substrate must be set to a period shorter than the period in which the chemical affinity is impaired.
On the other hand, according to the method for manufacturing an optical detection substrate of the present invention, a desired hydrophilic region can be easily provided on the surface of the substrate by forming a specific thin film on the surface of the substrate. Is also relatively easy. Moreover, even if the hydrophilicity imparted to the substrate by the method is affected by contaminants from the air, the effect is stably exhibited as long as the difference in contact angle with the hydrophobic region is maintained. Good handling of the substrate.

Also, when the hydrophilicity of the entire substrate surface is uniform, the spot shape and size should be controlled uniformly by managing the sample solution spot and the immobilization environment (humidity and / or temperature, etc.). Then, control by strict production management is required. Further, each time a substrate having a different surface property is used, it is necessary to reexamine the surface property and the spot condition, which is complicated.
On the other hand, since the substrate of the present invention has a surface structure in which fine hydrophilic regions are periodically provided as surface properties of the substrate itself, it can be spotted with good reproducibility regardless of the environment. In addition, there is an advantage that the necessity of adjusting the spot condition according to the environmental change is low.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these. In the examples, as the dichloropentafluoropropane (hereinafter referred to as R-225), product name AK-225 manufactured by Asahi Glass Co., Ltd. was used. As the platinum catalyst, a 3% by mass xylene solution of a Pt-divinyltetramethyldisiloxane complex was used. The NMR spectrum data was shown as an apparent chemical shift range, and the integral value was expressed as a ratio. The contact angle with respect to water was determined as an average value of three measured contact angles by placing three droplets on the substrate using a sessile drop method.

[Synthesis Example 1]
(Synthesis Example 1-1)
In a 500 mL glass reaction vessel equipped with a thermometer, a stirrer, and a Dimroth cooler, the purity by C ₈ F ₁₇ CH ₂ CH ₂ CH ₂ Br (gas chromatography (hereinafter referred to as GC)) is 84.3 mol. %)), 22.7 g of vanillin, 39.0 g of potassium carbonate, 6.87 g of tetrabutylammonium bromide and 170 g of acetonitrile were heated to 82 ° C. and refluxed for 1.5 hours. The reaction crude liquid was obtained.
450 g of R-225 was added to the resulting reaction crude liquid, and then washed twice with 300 g of distilled water. The solvent R-225 was distilled off to obtain 109 g of the following compound a. The purity by GC was 79.5 mol%, and the yield was 99.5%. The measurement results of ¹⁹ F-NMR and ¹ H-NMR are shown below.

¹⁹ F-NMR (solvent: CDCl ₃ ) δ (ppm): −81.3 (3F), −114.7 (2F), −122.4 (6F), −123.2 (2F), −123. 9 (2F), -126.6 (2F).
¹ H-NMR (solvent: CDCl ₃ ) δ (ppm): 2.13-2.42 (4H), 3.90 (3H), 4.16 (2H), 6.93-7.44 (3H) , 9.84 (1H).

(Synthesis Example 1-2)
In a 1 L glass reaction vessel equipped with a thermometer, a dropping funnel, a stirrer, and a Dimroth condenser, 109 g of compound a obtained in Synthesis Example 1-1 and 330 g of acetic acid were placed, and 128 g of fuming nitric acid and acetic acid were added. A solution mixed with 105 g was added dropwise at room temperature over 30 minutes. Then, it was further reacted for 3 hours to obtain a reaction crude liquid.
300 g of R-225 was added to the obtained reaction crude liquid, and then washed with 500 g of distilled water, 500 g of an 8% by mass aqueous sodium bicarbonate solution, and 500 g of distilled water. The solvent R-225 was distilled off to obtain 111.8 g of the following compound b. The purity by GC was 73.6 mol%, and the yield was 82.7%. The measurement results of ¹⁹ F-NMR and ¹ H-NMR are shown.

¹⁹ F-NMR (solvent: CDCl ₃ ) δ (ppm): −81.2 (3F), −114.7 (2F), −122.4 (6F), −123.2 (2F), −123. 8 (2F), -126.6 (2F).
¹ H-NMR (solvent: CDCl ₃ ) δ (ppm): 2.17-2.43 (4H), 3.99 (3H), 4.21 (2H), 7.40 (1H), 7.58 (1H), 10.44 (1H).

(Synthesis Example 1-3)
In a 300 mL glass reaction vessel equipped with a thermometer, stirrer, and Dimroth condenser, 26.8 g of compound b obtained in Synthesis Example 1-2, 3.3 g of hydrazine monohydrate, and 150 g of ethanol were added. The mixture was heated to 78 ° C. and refluxed for 3 hours to carry out the reaction. The crude product precipitated upon cooling to room temperature was collected by filtration and washed 5 times with 50 g of ethanol. After drying under reduced pressure, 17.1 g of the following compound c was obtained. The purity by NMR was 100 mol%, and the yield was 85.1%. The measurement result of ¹ H-NMR is shown.

¹ H-NMR (solvent: CDCl ₃ ) δ (ppm): 2.12-2.39 (4H), 3.95 (3H), 4.13 (2H), 5.81 (2H), 7.48 (1H), 7.55 (1H), 8.41 (1H).

(Synthesis Example 1-4)
In a 200 mL glass reaction vessel equipped with a thermometer, stirrer and Dimroth condenser, 16.8 g of compound c obtained in Synthesis Example 1-3, 10.9 g of manganese (IV) oxide and 100 g of chloroform were added. The mixture was stirred and reacted at room temperature for 3 hours to obtain a reaction crude liquid. The obtained reaction crude liquid was filtered, and the filtrate was washed with 100 g of an 8% by mass aqueous sodium bicarbonate solution and dried using 10 g of magnesium sulfate to obtain a reaction liquid.
In a glass container containing 1.8 g of 3-buten-1-ol, 0.36 g of a 53% mass tetrafluoroboric acid aqueous solution and 100 g of chloroform, the whole amount of the obtained reaction solution was put under cold water over 45 minutes. It was dripped. Furthermore, after making it react at room temperature for 3 hours, chloroform of the solvent was distilled off and it refine | purified by column chromatography and obtained 3.04g of the following compound d. The yield was 17.0%. The measurement result of ¹ H-NMR is shown.

¹ H-NMR (solvent: CDCl ₃ ) δ (ppm): 2.10-2.44 (6H), 3.62 (2H), 3.93 (3H), 4.13 (2H), 4.88 (2H), 5.04-5.18 (2H), 5.80-5.94 (1H), 7.25 (1H), 7.67 (1H).

(Synthesis Example 1-5)
In a 50 mL stainless steel pressure resistant reactor equipped with a PTFE inner bag, 4.60 g of compound d obtained in Synthesis Example 1-4, 2.76 g of triethoxysilane, 29 mg of Pt catalyst, and 15 g of toluene. And reacted at 100 ° C. for 1.5 hours to obtain a reaction crude liquid.
The solvent, toluene, was distilled off from the resulting reaction crude liquid and purified by column chromatography to obtain 2.24 g of the following compound e. The purity by NMR was 98 mol%, and the yield was 75.3%. The measurement result of ¹ H-NMR is shown. The absorption spectrum is shown in FIG.

¹ H-NMR (solvent: CDCl ₃ ) δ (ppm): 0.66 (2H), 1.20 (9H), 1.52-1.76 (4H), 2.11-2.42 (4H) 3.59 (2H), 3.78 (6H), 3.95 (3H), 4.14 (2H), 4.86 (2H), 97.30 (1H), 7.69 (1H).

[Pretreatment of glass substrate]
The surface of a 10 cm square 2 mm thick soda lime glass substrate was polished and washed with an abrasive containing cerium oxide fine particles, rinsed with pure water and air-dried to obtain a cleaned glass. In the following examples, the cleaned glass was used as a glass substrate.

[Reference Example 1]
In a 300 mL glass container, 1 part by mass of compound e obtained in Synthesis Example 1, 100 parts by mass of 2-propanol and 0.5 part by mass of 0.1N hydrochloric acid were added, and the hydrolysis reaction was performed at room temperature for 12 hours. To obtain a water-repellent composition e. 1 mL of the obtained water-repellent composition e was dropped onto a glass substrate, spin-coated at 3000 rpm for 20 seconds, washed with R-225, and air-dried to prepare sample e-1.
The contact angle with respect to water of the obtained sample e-1 was 108.8 degrees, and the contact angle with respect to water of the untreated glass substrate was 10 degrees or less.
Sample e-1 was irradiated with ultraviolet rays using a high-pressure mercury lamp, washed with R-225, and air-dried to produce sample e-2. When the irradiation amount was 22 mJ / cm ^2, the contact angle of Sample e-21 with water was 102.2 degrees.
In addition, when the total irradiation amount was 629 mJ / cm ^{2 by} repeating light irradiation and washing 12 times, the contact angle of Sample e-22 with water was 79.8 degrees.
When the sample e-1 is irradiated with ultraviolet rays so that the single dose is 650 mJ / cm ² , the contact angle of the sample e-23 with water is 90.9 degrees, and the same dose ( When the total irradiation amount was 3250 mJ / cm ^{2 by} repeating light irradiation and cleaning at 650 mJ / cm ² ) 5 times, the contact angle with respect to water of sample e-24 was 62.3 degrees.

[Reference Example 2]
For comparison, irradiation with ultraviolet rays and washing were repeated 12 times on an untreated glass substrate to make the total irradiation amount 629 mJ / cm ² , but the contact angle with water was 10 degrees or less.

[Example 1]
In Reference Example 1, the hydrophilic region 2 having the pattern shown in FIG. 1 is formed on the surface of the substrate 1 by selectively performing ultraviolet irradiation in the process of creating the sample e-1 to the sample e-24 using a photomask. Formed. The contact angle of the hydrophilic region 2 is 62.3 degrees, and the contact angle of the hydrophobic region 3 is 108.8 degrees.
The width (Wx) of the hydrophilic region 2 in the X direction was 100 nm, and the width (Wy) of the hydrophilic region 2 in the Y direction was 100 nm. The width (Sx) of the hydrophobic region 3 in the X direction was 50 nm, and the width (Sy) of the hydrophobic region 3 in the Y direction was 50 nm.
Therefore, the period of the hydrophilic region 2 in the X direction is 150 nm, and the period of the hydrophilic region 2 in the Y direction is 150 nm.

On the substrate 1 obtained above, ink water colored with ink as a sample solution was spotted. That is, spots were formed by applying ink water to the tip of a stainless pin having a diameter of 100 μm and bringing it into contact with the surface of the substrate 1 for transfer. Similarly, 100 spots were formed on the same substrate 1 while being separated from each other.
Thereafter, after the ink water on the substrate 1 was dried, the shape of each spot indicated by the ink remaining on the substrate 1 was observed with an SEM (scanning electron microscope). Each spot was a circular shape that was almost a perfect circle. The ink density in the spot was uniform. The major axis of each spot was measured. The results are shown in Table 1 below.

[Comparative Example 1]
As a comparative example, the same ink water as in Example 1 was spotted on the surface of sample e-1 (contact angle 108.8 degrees) on which the water repellent thin film was formed on the entire surface of the substrate, and the same was true for the obtained spots. When observed, it was indefinite, such as punctate or crescent. Further, the major axis could not be measured due to the irregular shape. The results are shown in Table 1 below.

[Comparative Example 2]
As a comparative example, ink water similar to that in Example 1 was spotted on an untreated glass substrate (contact angle of 10 ° or less), and each spot obtained was similarly observed. Each spot had a substantially circular shape, but the closer to the outer peripheral part than the central part, the deeper the ink, and the non-uniform ink distribution. The major axis of each spot was measured. The results are shown in Table 1 below.

  From the results in Table 1, in the optical detection substrate obtained in Example 1, a spot that is almost a perfect circle is obtained, the spot shape is highly uniform, and there is little variation in size. In addition, the distribution of the ink in the spot is uniform, and from this, when the sample solution contains a material to be fixed, it is estimated that the uniformity of the material to be fixed is good.

It is a top view showing one embodiment concerning the present invention. It is a top view which shows the modification based on this invention. It is a top view which shows the modification based on this invention. 3 is an ultraviolet-visible light absorption spectrum of compound e in Synthesis Example 1.

Explanation of symbols

1 Substrate 2 Hydrophilic region 3 Hydrophobic region

Claims (6)

A substrate used in a method for detecting detection light emitted from a substance fixed on a substrate,
A substrate for optical detection, wherein hydrophilic regions are periodically provided on the surface of the substrate, and the period of the hydrophilic regions is 5000 nm or less and smaller than the diameter of the fixing region of the substance.   The optical detection substrate according to claim 1, wherein a period of the hydrophilic region is 1000 nm or less.   The substrate for optical detection according to claim 1, wherein the hydrophilic region is subjected to a surface treatment that promotes fixing of the substance.   The optical detection substrate according to any one of claims 1 to 3, wherein the hydrophilic regions have the same period in two different repeating directions perpendicular to the thickness direction of the substrate.   The optical detection substrate according to any one of claims 1 to 4, wherein a diameter of a fixing region of the substance is 50 to 500 µm. A method for producing a substrate for optical detection according to any one of claims 1 to 5, wherein a thin film whose affinity with water is changed by irradiation with active energy rays on the surface of the substrate is formed. And a method for selectively irradiating the thin film with active energy rays.

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