JPH0640813B2 - Incubator - Google Patents

Description

TECHNICAL FIELD The present invention relates to an incubator, and more particularly to an incubator at least a part of which is a fluorine-containing molten resin.

[Prior Art] Since an incubator provides an environment for culturing cells, tissues, organs, etc. of animals and plants, or microorganisms, it satisfies various properties such as light permeability, low moisture permeability, durability, and chemical resistance. Required to do so. As an incubator satisfying these properties, a glass incubator or the like has been conventionally used. The opening of a glass incubator or the like is generally closed with a cotton plug, a rubber plug with a cotton plug, an aluminum foil, a paper stopper, or the like.
In addition, a stopper in which a rubber stopper body is hollowed out and the opening is closed with a sheet of open-cell foam has been developed.

However, in these stoppers, caps, etc., various bacteria may enter with the inflow of air due to temperature change. In addition, the shape of the incubator is limited to bottles, test tubes, etc. from the viewpoint that glass is fragile and processability is high. Therefore, there is a demand for an incubator which has a light-transmitting property comparable to that of glass, ensures good air permeability without invasion of various bacteria, and is not broken.

[Object of the Invention] A first object of the present invention is to provide an unbreakable incubator which does not allow invasion of germs, has good air permeability and low moisture permeability, and has light transmittance comparable to that of glass. Especially.

A second object of the present invention is to provide an incubator whose shape can be freely designed.

[Constitution of the Invention] The incubator of the present invention is characterized in that at least a part thereof is made of a fluorine-containing molten resin.

As the fluorine-containing molten resin, tetrafluoroethylene /
Hexafluoropropylene copolymer (hereinafter referred to as FEP polymer), tetrafluoroethylene / perfluoro (alkyl vinyl ether) copolymer (hereinafter referred to as PFA polymer), especially tetrafluoroethylene / perfluoro (propyl vinyl ether) copolymer Polymers, ethylene / tetrafluoroethylene / other fluoroolefin copolymers, fluorinated acrylate resins [eg, formula: (In the formula, m is an integer of 0 to 5, and X represents a fluorine or trifluoromethyl group.) A polymer composed of a repeating unit.

Since such a fluorine-containing molten resin has low moisture permeability and also has gas permeability necessary for culture, it is suitable for culture requiring air permeability.

The incubator may consist of one part, or two separable parts, such as an incubator body and a closure, or an incubator body and a metal or resin frame housed inside the body. It may consist of three or more parts. The term “sealing tool” includes all forms of devices such as a cap screwed from the outside of the opening, such as a cap, and the fluorine-containing molten resin film itself for closing the opening of the incubator.

In the present specification, at least a part of the incubator includes the entire incubator, and the incubator may be made of only the fluorine-containing molten resin film. Alternatively, the incubator may be made of, for example, a film obtained by joining a fluorine-containing resin film and another resin film. When the incubator has a sealing tool, at least a part of only the sealing tool may be made of the fluorine-containing molten resin.

The thickness of the fluorine-containing molten resin film is usually 2 to 2500.
μm, preferably in the range of 4 to 100 μm.

Since the fluorine-containing molten resin can be easily processed by means such as welding, heat bonding, adhesive bonding, and molding, an incubator having an arbitrary shape can be easily manufactured.

The incubator of the present invention can be used in the same fields where conventional glass incubators have been used.

Hereinafter, some aspects of the present invention will be described with reference to the accompanying drawings. However, the present invention is not limited to the embodiments shown in the drawings.

1 to 5 are perspective views showing the shape of the incubator of the present invention. The films constituting the incubators 1, 2, 3, 4 and 5 may be films composed only of the fluorine-containing molten resin, or may be laminated films having a layer of the fluorine-containing molten resin.

Since the envelope-type incubator (FIG. 1) can be arranged in a stacked state, it does not take up a lot of space for culturing, and is particularly useful for culturing a large number of small cultures. In addition, since the quadrangular prism type and the hexagonal column type incubators can be arranged without a gap, it is possible to effectively use a place such as a culture shelf.

After placing the medium and planting the culture, the upper parts of these incubators are sealed. The sealing may be performed by any method as long as it can completely prevent the inflow of outside air from the opening and the mixing of various bacteria, and can be performed by, for example, a heat bonding method or a method using a sealing tool. When using a sealer, use the incubator 1
One of the walls, preferably the upper wall, may be fitted with a stopper such as is attached to a conventional plastic culture bottle, and a culture medium or the like may be inserted from the stopper and then sealed with a cap. The stopper and cap are preferably made of resin.

It is also possible to form an appropriate opening in one wall of the incubator, put a medium or the like therein, and then close the opening with a film or sheet piece slightly larger than the opening. The film or sheet piece may be attached in advance by adhering or fusing a part of it to the wall around the opening so as to overlap the opening.

FIG. 6 is a cross-sectional view of the sealing device according to the present invention, at least a part of which is made of fluorine-containing molten resin. A wall made of a fluororesin film 13 is attached to the main body 12 of the sealing tool at the upper part of the sealing tool 11 which is a cap for an incubator. The cap 11 is a packing 14 made of an elastic material.
Also has.

EXAMPLES Reference Example 1 Light Transmission For a 25 micron or 250 micron thick FEP polymer and a 25 micron thick PFA polymer,
The transmittance of light having a wavelength of 1.0 micron or less was examined. The results are shown in FIGS. 7 and 8, respectively.

It can be seen that the fluorine-containing molten resin used in the present invention has excellent light transmittance in the infrared to ultraviolet wavelength range.

Reference Example 2 Gas Permeability According to the method specified in ASTM D 1434, FE
Gas permeability coefficients of nitrogen, oxygen and carbon dioxide (unit: cm ³ · cm / cm ² · s) for P and PFA polymers
ec • atm) was measured. The results are as follows.

FEP Polymer Nitrogen 120x10 ^-10 Oxygen 370x10 ^-10 Carbon Dioxide 970x10 ^-10 PFA Polymer Nitrogen 140x10 ^-10 Oxygen 480x10 ^-10 It is understood that the fluorine-containing resin used in the present invention has good air permeability.

Reference Example 3 Water vapor transmission rate FEP polymer (thickness 45
Micron) and PFA polymer (thickness 45 microns) were measured for water vapor transmission. The results are as follows.

FEP polymer 2.0 g / m ² · 24 hours PFA polymer 3.1 g / m ² · 24 hours It can be seen that the fluorine-containing molten resin used in the present invention has low moisture permeability.

Example 1 One sheet of NEOFLON FEP (manufactured by Daikin Industries, Ltd., tetrafluoroethylene / hexafluoropropylene copolymer) having a thickness of 100 μm was heat-bonded with an impulse sealer to obliquely cut an upper corner in an envelope type. Two mold incubators were manufactured. The width of the heat-sealed parts on the sides and bottom is 5 mm, and the dimensions of the incubator are 15 cm in the vertical long side, 8 cm in the short side, and 4 cm in the horizontal upper side (opening).
The lower side was 8 cm. After sterilizing these incubators by an autoclave, 80 ml of modified Vacin and Went liquid medium of PH5.3 shown in Table 1 was injected through each opening, and 60 explants were inoculated respectively. Then, the air on the upper side was expelled, and the incubator was sealed by sealing with a closer (manufactured by Spectrum Medical Industries) and thermally bonding the outside. The explants used were the Cymbidium Shougirl "Husky".
After culturing the protocomb-shaped spheres (PLB) of "Honey") for 1 month or more after division, shoots were undifferentiated and PLB having an average diameter of 2.4 mm and an average fresh weight of 7.3 mg were divided into four vertically. It was

75 at 25 ° C and 900 lux (16 hours / day)
When cultured for one day, the number of surviving PLBs in the two incubators was 113 in total, and the average number of PLBs grown from one explant was 11.9.

Example 2 except that the NEOFLON FEP film was changed to NEOFLON PFA (a tetrafluoroethylene / perfluoro (alkyl vinyl ether) copolymer, manufactured by Daikin Industries, Ltd.) film, the number of incubators was set to 3, and the number of culture days was set to 65 days. The procedure of Example 1 was repeated. The total number of surviving PLBs was 163 in the three incubators, and the average number of proliferating PLBs was 15.7.

Example 3 A rectangular prism-shaped incubator consisting of only a 25 μm thick neoflon FEP film was manufactured. Bend the film and heat seal both ends with a width of about 5 mm. After forming a square columnar cylinder, fold the two opposite sides of one opening of the square columnar cylinder in the middle of the cylinder. , And the other two sides were heat-sealed with a width of about 5 mm from the end. The inner dimensions of this square column type container are 7.5 cm x 7.5 cm at the bottom,
The height was 20 cm. The outer dimensions are 7.5 cm x 7.5 cm x so that the shape of this square column type container does not collapse.
A 10.5 cm stainless steel frame (diameter 2 mm) was inserted into the inside of the rectangular prism type container. After injecting 100 ml of a liquid medium of PH5.0 shown in Table 2 into an incubator, it was sterilized by autoclaving. In addition to this, Cypbidium Valley Flower “Cch
After planting 9 seedlings with 2 to 3 true leaves of erry Ripe ") and a leaf length of about 2 cm at even intervals,
The opening was sealed with an impulse sealer and cultured at 25 ° C. and 2000 lux (16 days / day) for 4 months. When the size and morphology of the seedlings after culturing were observed, no abnormalities were observed and they showed favorable growth.

Example 4 A glass beaker having a diameter of 8 cm and a height of 13 cm was used, and the opening was covered with a 4 μm-thick NEOFLON PFA film.
The same culture as in Example 3 was performed except that the gap between the film and the beaker was completely sealed with a vinyl tape. Observation of the size and morphology of the seedlings after culturing revealed no abnormalities and showed favorable growth.

Example 5 Peptone 2% in a 500 ml Erlenmeyer flask (caliber 29 mm),
Medium containing 2% yeast extract and 4% glucose (PH
6.0) 100 ml was put, the opening was covered with aluminum foil, and then autoclave sterilized, and Saccharomyces cerevisiae IFO 0309
The strain was inoculated with 1 platinum loop. Next, replace the aluminum foil and cover the opening with a 4 μm thick NEOFLON PFA film.
After completely sealing the gap between the film and the glass surface with a vinyl tape, rotary shaking culture was carried out at 35 ° C. and 200 rpm for 48 hours. As a result, it was confirmed that the bacterial cells grew smoothly. Also, no contamination of various bacteria was observed.

Example 6 The strain was Rhodotorula mu
cilaginosa) Example 5 except that it is changed to AHU 3946 strain
The above procedure was repeated.

As a result, it was confirmed that the bacterial cells grew smoothly. or,
No bacterial contamination was observed.

Comparative Example 1 Example 1 was repeated except that two 300 ml glass Erlenmeyer flasks were used as an incubator and the openings were covered with aluminum foil.
The same procedure was repeated. The number of surviving PLBs was 102 in total in the two flasks, and the average number of proliferating PLBs was 9.
It was 0.

[Effects of the Invention] The features of the incubator of the present invention are as follows.

Since the breathability of the fluorine-containing molten resin film is not due to the fine pores, there is no risk of miscellaneous bacteria entering the incubator. The low moisture permeability prevents the medium from drying.

It has a light transmission property suitable for the growth of a culture.

Since it has heat resistance, it is normally sterilized (121 ℃ x 15 minutes)
Can withstand enough.

[Brief description of drawings]

FIG. 1 is a perspective view of an envelope type incubator of the present invention, FIG. 2 is a perspective view of a cylindrical type incubator of the present invention, and FIG. 3 is a perspective view of a square column type incubator of the present invention. FIG. 4 is a perspective view of a regular tetrahedron incubator of the present invention, FIG. 5 is a perspective view of a hexagonal prism-shaped incubator of the present invention, and FIG. 6 is a cross-sectional view of an incubator cap of the present invention. FIG. 8 is a graph showing the light transmittance of the tetrafluoroethylene / hexafluoropropylene copolymer, and FIG. 8 is a graph showing the light transmittance of the tetrafluoroethylene / perfluoroalkyl vinyl ether copolymer. 1, 2, 3, 4, 5 ... Incubator, 11 ... Incubator cap, 12 ... Cap body, 13 ... Fluorine-containing molten resin film, 14 ... Packing.

Claims (3)

[Claims] 1. An incubator characterized by comprising at least a part of a fluorine-containing molten resin. 2. The incubator according to claim 1, wherein the fluorine-containing molten resin is a tetrafluoroethylene / hexafluoropropylene copolymer. 3. The incubator according to claim 1, wherein the fluorine-containing molten resin is a tetrafluoroethylene / perfluoro (alkyl vinyl ether) copolymer.