JPS63151913A - Immersion oil for microscope - Google Patents

Abstract

PURPOSE:To obtain immersion oil for a microscope having low fluorescence and other satisfactory properties by blending specified phthalic esters as base oil with a specified liq. polymer, etc. CONSTITUTION:Phthalic esters are blended with (A) a liq. diene polymer and liq. paraffin or halogenated paraffin or with (B) polybutene to obtain immersion oil for a microscope. Phthalic esters which are liq. at ordinary temp. and pressure or mixed phthalic esters which are liq. at ordinary temp. and pressure are used as the phthalic esters without any special restriction. A liq. diene polymer having 300-25,000, preferably 500-10,000 number average mol.wt. is generally used as the liq. diene polymer. Therefore, phthalic esters [(C) component], diene polymer [(A1) component] and liq. paraffin [(A2) component] or halogenated paraffin [(A3) component] are blended or the component (C) and polybutene [(B) component] are blended to obtain the titled oil.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an immersion oil for microscopes, and more particularly to an immersion oil that has low fluorescence and is particularly suitable for use in fluorescence microscopes.

[Conventional technology]

One method of increasing the magnification of a microscope is to use immersion oil, which increases the numerical aperture of the objective lens. The immersion oil used in this case is one consisting of benzylbutyl phthalate and chlorinated paraffin (U.S. Pat. No. 4,465
, 621), and one consisting of a liquid diene polymer and liquid paraffin (Japanese Patent Application Laid-Open No. 61-7438).

However, although these immersion oils have almost all the properties required of immersion oils for microscopes, such as refractive index, Abbe's number, viscosity, and resolution, their fluorescence is not comparable when measured using a spectrophotometer. It has drawbacks such as being harsh on fluorescent light, and is not satisfactory as an immersion oil for fluorescence microscopes, which is generally used for observing objects that emit fluorescence.

[Problem that the invention seeks to solve]

It is an object of the present invention to solve the above-mentioned problems and to provide an immersion oil for microscopes that has low fluorescence itself and has good other properties, and is particularly suitable as an immersion oil for fluorescence microscopes. It is something to do.

[Means for solving the above problems]

Based on the above circumstances, the inventor of the present invention has conducted repeated studies to develop an immersion oil that has low fluorescence and is also excellent in other properties. It has been discovered that the above object can be achieved by blending a specific liquid polymer, etc., and the present invention has been completed based on this knowledge.

That is, the present invention provides phthalate esters and (A
The present invention provides an immersion oil for a microscope, which is characterized by comprising: (a) a liquid diene polymer and a liquid paraffin or a halogenated paraffin; or (B) a polybutene.

The phthalate esters used in the present invention are not particularly limited as long as they are phthalate esters that are liquid at room temperature and pressure, or mixed phthalate esters that are liquid at room temperature and pressure (usually, the following General formula (Formula [1]), R3゜(Here, R1, R2, R3 are each an alkyl group,
A hydrocarbon group such as a cycloalkyl group, an alkenyl group, an aralkyl group, or an aralkyl group; a halogen atom, a hydroxyl group, an alkoxy group, an alkoxycarbonyl group,
Represents a substituted hydrocarbon group substituted with an acyloxy group. n represents an integer of 0 to 4, preferably 0. The p+, R2 is c+ -Cm. alkyl group of C1 to CIO, cycloalkyl group of C1 to CIO, aryl group of 06 to CI+1, C?
~C1! Aralkyl groups such as methyl, ethyl, propyl, butyl, hexyl, octyl, decyl, undecyl, dodecyl, tridecyl, benzyl and the like are particularly preferred. As R3,
Lower alkyl groups such as methyl and ethyl groups are preferred.

The substitution position of “-COOR” in the formula [1] is as follows:
Those in the ortho position of the -COOR' group are preferred.

The groups pl, R2, and R1 may be of the same type or different types. )
Or, the following general formula (Formula [2]), R'a (where R5 is a divalent hydrocarbon group such as an alkylene group, an alkenylene group, an arylene group; 2 substituted with a halogen atom, an alkoxy group, etc.) represents a substituted hydrocarbon group.R
4 represents the same group as R3 in the above formula [1], and represents an integer from 0 to 4, preferably 0. The two -COO- groups in the formula [2] are preferably substituted at ortho positions. In the formula [2], -R8- is an ethylene group (-CLCL-), a propylene group (C)I (CL)-C)It-), a trimethylene group,
Tetramethylene group, methylene can be formed. R4 is preferably a lower alkyl group such as a methyl group or an ethyl group. ) or a mixture of two or more phthalate esters, which are liquid at normal temperature and pressure.

Among these phthalate esters, preferred ones are shown merely for the purpose of illustration and not limitation, for example, 1, 2
-Dimethyl benzenedicarboxylate, diethyl 1,2-benzenedicarboxylate, dipropyl 1,2-benzenedicarboxylate, di-n-butyl 1,2-benzenedicarboxylate, diisobutyl 1,2-benzenedicafrefonate, 1,
Di-n-hexyl 2-benzenedicarboxylate, diisohexyl 1.2-benzenedicarboxylate, di-n-octyl 1.2-benzenedicarboxylate, diisooctyl 1,2-benzenedicarboxylate, di-n-decyl 1.2-benzenedicarboxylate , diisodecyl 1,2-benzenedicarboxylate, di-n-dodecyl 1,2-benzenedicarboxylate, 1
．． Diisodecyl 2-benzenedicarboxylate, di-n,-)lysosyl 1,2-benzenedicarboxylate, diisotridecyl 1,2-benzenedicarboxylate, benzylmethyl 1,2-benzenedicarboxylate, benzylethyl 1,2-benzenedicarboxylate, Examples include benzylpropyl 1,2-benzenedicarboxylate, benzyl n-butyl 1,2-benzenedicarboxylate, and benzylisobutyl 1,2-benzenedicarboxylate. In addition, these phthalate esters can be used as a single compound as long as they are liquid at normal temperature and pressure, or two or more types can be used as a liquid mixture. can.

The liquid diene polymer used in the present invention is not particularly limited, but usually has a number average molecular weight of 300 to 25,000,
Preferably, a liquid diene polymer having a molecular weight of 500 to 10,000 is used.

Examples of these liquid diene polymers include diene polymers and diene copolymers having 4 to 12 carbon atoms, and copolymers of these diene monomers and α-olefinic addition polymerizable monomers having 2 to 22 carbon atoms. There is. Specific examples include butadiene homopolymer, isoprene homopolymer, butadiene-styrene copolymer, butadiene-isoprene copolymer, butadiene-acrylonitrile copolymer, butadiene-2-ethylhexyl acrylate copolymer, butadiene-n-octadecyl acrylate copolymer, and the like. .

Here, the liquid diene polymers such as these exemplary liquid diene polymers may not contain a hydroxyl group, may have a hydroxyl group within the molecule and/or at the end of the molecule, or , these may be a mixture, and the content of hydroxyl groups is usually 0.1 to 10 mm!
/g, preferably 0.3 to 7 milliequivalents/g, can be particularly preferably used.

These hydroxyl group-containing liquid diene polymers are produced by, for example, converting a conjugated diene monomer into a liquid reaction medium in the presence of hydrogen peroxide.
It can be produced by heating reaction.

Note that these liquid diene polymers may be used alone or in combination of two or more.

The liquid paraffin used in the present invention is not particularly limited as long as it is so-called liquid paraffin, but usually,
For example, relatively light lubricating oil fractions such as spindle oil fractions, refrigeration machine oil fractions, machine oil fractions, dynamo oil fractions, turbine oil fractions, etc., preferably spindle oil fractions, are washed with sulfuric acid, treated with white clay, etc. Liquid hydrocarbon oils mainly composed of alkylnaphthenes obtained through highly refined oils are preferred.

Among these, the density is 0.860-0.905
A material of about g/cd can be particularly preferably used.

Specifically, for example, industrial liquid paraffin, pharmacopeia liquid paraffin, etc. can be mentioned.

The halogenated paraffin used in the present invention is a liquid halogenated paraffin or a liquid mixed halogenated paraffin. Specifically, liquid halogenated paraffins such as fluorinated paraffins, chlorinated paraffins, brominated paraffins, and iodinated paraffins, or fluorine, chlorine, bromine,
Examples include liquid composite halogenated paraffins or liquid mixed halogenated paraffins containing at least two types of halogen atoms selected from iodine.

Among these, chlorinated paraffins are preferred. The above halogenated paraffin usually has a halogen content of 1
0 to 80% by weight, preferably 20 to 70% by weight, acid value 0.01 to 0.50 KOHmg/g, viscosity 0.
5-40,000 bois (25℃), specific gravity is 1.10
~1.80 (25℃), chromaticity 5.50~350 (A
PHA) are preferably used, and liquid chlorinated paraffins having these properties are particularly preferred.

Note that if the properties of the halogenated paraffin such as the chlorinated paraffin or a mixture thereof are not within the range of properties described above, the fluorescence of the obtained immersion oil may become high, and the welding ratio, Atsube number, kinematic viscosity, In some cases, the properties such as transparency and chromaticity become poor.

The polybutene used in the present invention is polyisobutene obtained by polymerizing isobutyne or copolymerizing isobutyne with a small amount of n-butene, or a mixture thereof, and is a liquid polybutene having a number average molecular weight of 100 to 2,500. be. These polybutenes can be used as liquid polybutenes having various viscosities or liquid mixtures thereof, but usually have a viscosity of 4.5 centistokes (98.9°C) or more, 100 centistokes (98.9°C) or more.
Low viscosity polybutene (B+) with a viscosity of less than 8,9°C) and a viscosity of 100 to 4600 centistokes (98,9°C)
A liquid polybutene is obtained by blending high viscosity polybutene (B2) in a range of 20 to 70% by weight of the above (B1) and 80 to 30% by weight of the B2, and the resulting immersion oil It can be particularly suitably used in terms of properties such as kinematic viscosity.

These polybutenes are, for example, the residue obtained by extracting butadiene from the C4 fraction during naphtha decomposition, that is, isobutyne, a small amount of n-butene such as 1-butene, and an isobutyne mixture containing n-butane as a main component. The reaction conditions are usually 5 to 130°C and 1 to 22 kg/adG using aluminum chloride as a raw material and aluminum chloride as a catalyst. Polybutenes of various viscosities can be produced by adjusting the temperature and polymerizing.

The immersion oil for microscopes of the present invention comprises the phthalate ester [
(C) component], the liquid diene polymer ((AI) component), and the liquid paraffin ((A2) component) or the halogenated paraffin [(A3) component], or (C) It can be obtained by blending the component and the polybutene [component (B)].

The blending ratio of each of these components is as follows.

That is, the (C) component and the (A +)+ (A
the law of nature .

The ratio of (A,) is per 100 parts by weight of component (C).
The total amount of Aυ, (A2) and (A3), that is, the above (A)
The amount of the component is usually 50 to 300 parts by weight, preferably 70 parts by weight.
~200 parts by weight, and further the above (A,) and the above (AZ
) and the above (A, ), (A1), (A2) and (
When the total amount of A,) is 100% by weight, (AI
) is 20 to 70% by weight, and the total amount of (A2) and (A, ) is 80 to 30% by weight. Here, (A8) and (A,
) is an arbitrary value, i.e. (Az)/
It can be set in the range of ((Az) + (As)) - 1 to 1, but usually (Ax) / ((Ax) + (Az
))=O or one having this value of 1 can be particularly preferably used from the viewpoint of simplifying the manufacturing process.

Here, the component (C), the component (A,), and the component (A2)
And/or if the blending ratio with the component (A,) is not within the above range, the fluorescence of the obtained immersion oil may become high, and various properties such as pressure welding ratio, number of pistons, kinematic viscosity, etc. may deteriorate. There may be cases where

On the other hand, the ratio of the component (C) to the component (B) is (
C) 1. per 100 parts by weight of component. (B) Ingredients are usually 20%
~150 parts by weight, preferably 50-100 parts by weight.

Here, if the blending ratio of the component (C) and the component (B) is not within the above range, the fluorescence of the obtained liquid man oil may become high, the pressure welding rate, the number of pimples, the kinematic viscosity, etc. characteristics may deteriorate.

In the method of this invention, the above (C), (AI), (A
2), the order of blending (A,) and the above (C) and the above (
B) Or, there is no particular restriction on the order of blending the above (C), the above (B1), and the above (B), and at the same time or
They can be combined step by step in various orders.

Further, there is no particular restriction on the method of blending, and a method of blending by stirring and mixing at around room temperature is usually suitably used.

In the manner described above, the immersion oil for microscopes of the present invention can be obtained. If necessary, additives, compounding agents, etc. used in immersion oils for microscopes such as ordinary immersion oils for fluorescence microscopes can be added to this immersion oil for microscopes.

The immersion oil for microscopes produced as described above can be suitably used as an immersion oil for ordinary microscopes, especially as immersion oil for fluorescence microscopes.

〔Example〕

Production of Polybutene 6 (Production Example-1) Production Example 500 of Low Viscosity Polybutene 5 g of aluminum chloride was placed in the autoclave, the pressure was reduced using a vacuum pump, and 50 g of n-butane was added as a solvent. After stirring to sufficiently disperse aluminum chloride,
200g of raw materials (Isobutyne 42%, 1-
A mixture of 2.18% by weight of butene and 40% by weight of n-butane) was added dropwise over 20 minutes and reacted at 30°C for 2 hours.

After the reaction is completed, unreacted gas and n-butane are removed,
300 g of water was added under water cooling to decompose aluminum chloride. Next, it was extracted three times with 500 ml of N-hexane, and the n-
Hexane was distilled off to obtain 80 g of polybutene.

Note that under these reaction conditions, the viscosity is 38 centistokes (
A low viscosity polybutene with a temperature of 98.9° C.) was obtained.

(Production Example-2) Production side of high viscosity polybutene -
Example 1 was carried out in the same manner as in Example 1 except that the reaction temperature was changed to 15°C. Under these conditions, the viscosity is 290 centistokes (
Approximately 80 g of high viscosity polybutene (98.9°C) was obtained.

(Examples 1 to 4 and Comparative Examples 1 and 2) Each component shown in Table 1 was blended in the indicated amount, and 10
A microscope immersion oil was prepared by stirring and mixing for several minutes.

Various evaluations described below were performed on these items.

The results are shown in Tables 1 and 2.

Evaluation method in Table 2 Refer to the section on low fluorescence In Table 1, in the evaluation column, the mark ◯ indicates good or suitable, and the mark X indicates poor or inappropriate. However, regarding the evaluation of appearance, Q mark indicates no cloudiness, and Δ mark indicates slight cloudiness.

Moreover, *1 to *7 in Table 1 are as follows.

*1 Number average molecular weight 1600, viscosity 750 centipoise (25°C) *2 Number average molecular weight 2800, hydroxyl group content 0.80 milliequivalents/g *3 Lightco, manufactured by Chemical Co., Ltd., Pentol *4 Chlorinated n-paraffin, chlorine content 59.3% by weight Acid value 0.08 XOTImg/g *5 Obtained by the method of Production Example-1 above.

Viscosity 38 centistokes (98.9°C) *6 Obtained by the method of Production Example-2 above.

Viscosity: 290 centistokes (98.9°C) *7 High viscosity, not very desirable.

Review jL beggars 1 [! (n i!'), 77 he number ("It3)
All conformed to JIS K2101.

School skin (25℃) Compliant with JIS K2283.

lonesomeness Fluorescence microscopes use fluorescence as a light source to excite An ultra-high-pressure mercury lamp that emits ultraviolet light was used.

The excitation light used in this case includes U excitation, (2) excitation, B excitation, and G excitation depending on the length of the wavelength. For each excitation light, immersion oil, which generates a small amount of fluorescence, is desirable for a fluorescence microscope.

Table 2 shows the fluorescence intensity (relative intensity) for each excitation light.

(Measured by Fluorescence Spectrophotometer FP-5504 manufactured by JASCO ■) lJJ Raw JTS C2201 "Electrical Insulating Oil" 12. According to the evaporation test, a test was conducted at 30°C for 24 hours and evaluated in the following two stages. .

” Take the sample in a clean glass container and We investigated whether there was any

Cloudy None－・−・−・○ Some presence−・−−−−−
-△Bush Mosquito i Evaluation was made in the following two stages based on the results of the following light irradiation test and heat deterioration test, as well as changes in refractive index, number of pimples, and hue before and after the test.

A certain amount (40±0.5g) of the sample was placed in a Petri dish (φ9cm).
), and the light source (highlight white FL20W manufactured by Matsushita Electric Industrial ■) was used, and the distance between the lamp and the sample was set at 1.
5 cm) for a certain period of time (24, 72, 12
0 hours) Changes in refractive index after irradiation were observed. No change was evaluated as good (O).

A certain amount (40 ± 0.5 g) of the sample was taken into a 50-piece Erlenmeyer flask with a stopper, kept in a constant temperature bath at a constant temperature (40, 70°C) for 24 hours, and then the refractive index was determined. , the number of Atsubes, and changes in hue were observed. No change was evaluated as good (◯).

The criteria for evaluation are the same as in the weather resistance section above.

The presence or absence of corrosivity was investigated by measuring the total acid value of blood (JIS K 2501) and the effect on smear dye (JIS K 2400). Absence was marked as ○, and presence was marked as x.

Contrast Contrast was evaluated in the following two stages by looking at black and white lines cut on a black and white plate coated with chromium using a microscope using the immersion oil.

five-fold direction Evaluation was made in the following two stages based on the refractive index.

chromatic aberration Evaluated in the following two stages depending on the Atsube number. Ta.

Evaluation was made in the following two stages based on skin permeability (JIS K 0115).

〔Effect of the invention〕

According to the present invention, the immersion oil itself has low fluorescence and has good other properties such as refractive index, Abbe's number, viscosity, and resolution, and is therefore particularly excellent as an immersion oil for fluorescence microscopy. immersion oil can be provided.

Idebetsu Idemitsu Petrochemical Co., Ltd.

Claims (1)

[Scope of Claims] 1. An immersion oil for a microscope characterized by comprising a phthalate ester and (A) a liquid diene polymer and liquid paraffin or halogenated paraffin, or (B) polybutene. 2. The immersion oil for a microscope according to claim 1, wherein the phthalate ester is a liquid phthalate ester. 3. The immersion oil for a microscope according to claim 1 or 2, wherein the phthalate ester is a 1,2-benzenedicarboxylic acid ester. 4. The ratio of phthalate esters and polybutene is 20 to 20 parts by weight of polybutene per 100 parts by weight of phthalate ester.
150 parts by weight of the immersion oil for a microscope according to any one of claims 1 to 3. 5. Polybutene has a viscosity of 4.5 centistokes (98
．． 20 to 70% by weight of a low viscosity polybutene having a viscosity of at least 9°C and less than 100 centistokes (98.9°C) and a viscosity of 10
The immersion oil for a microscope according to any one of claims 1 to 4, which is a polybutene consisting of 80 to 30% by weight of high viscosity polybutene having a viscosity of 0 to 4,600 centistokes (98.9°C). 6. The blending ratio of phthalate esters and the above (A) is
The immersion oil for a microscope according to any one of claims 1 to 3, wherein the amount of (A) is 50 to 300 parts by weight per 100 parts by weight of the phthalate ester. 7. The above (A) is 20 to 70% by weight of a liquid diene polymer
The immersion oil for a microscope according to any one of claims 1 to 3 or 6, comprising 80 to 30% by weight of liquid paraffin. 8. The above (A) is 20 to 70% by weight of a liquid diene polymer
The immersion oil for a microscope according to any one of claims 1 to 3 or 6, comprising 80 to 30% by weight of halogenated paraffin. 9. The liquid diene polymer has a number average molecular weight of 300 to
The immersion oil for a microscope according to any one of claims 1 to 3 and 6 to 8, which is a liquid diene polymer of 25,000%. 10. Claims 1 to 3, 6, and 8, in which the halogenated paraffin is a chlorinated paraffin
Immersion oil for microscopes according to any one of Item 1 and Item 9.