WO2012127650A1 - Dispositif de mesure de turbidité - Google Patents

Dispositif de mesure de turbidité Download PDF

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Publication number
WO2012127650A1
WO2012127650A1 PCT/JP2011/056955 JP2011056955W WO2012127650A1 WO 2012127650 A1 WO2012127650 A1 WO 2012127650A1 JP 2011056955 W JP2011056955 W JP 2011056955W WO 2012127650 A1 WO2012127650 A1 WO 2012127650A1
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WO
WIPO (PCT)
Prior art keywords
turbidity
light
culture
culture tank
optical path
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Application number
PCT/JP2011/056955
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English (en)
Japanese (ja)
Inventor
石川　陽一
宏幸 釣井
勉 妙圓薗
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エイブル株式会社
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Priority to PCT/JP2011/056955 priority Critical patent/WO2012127650A1/fr
Publication of WO2012127650A1 publication Critical patent/WO2012127650A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G01N21/51—Scattering, i.e. diffuse reflection within a body or fluid inside a container, e.g. in an ampoule
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G01N21/53—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke
    • G01N21/534—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke by measuring transmission alone, i.e. determining opacity

Definitions

  • the present invention relates to an apparatus for measuring the turbidity of a culture solution, and more particularly, to a turbidity measurement apparatus for optically measuring the turbidity of a culture solution in a culture tank from outside the culture tank through a tank wall. .
  • the concentration (number) of cells or cells in the culture solution or the degree of proliferation thereof is measured for culture management.
  • the turbidity of the culture solution may be used as an index when managing the culture state.
  • a turbidity sensor probe
  • the turbidity of the culture solution is optically measured. Is done.
  • a turbidity sensor irradiates a liquid with light from a light source of a light emitting unit, and measures turbidity from the amount of light received by a light receiving unit for detection at that time. And when calculating the concentration of microbial cells and cells from the turbidity of the culture solution, create a standard curve (calibration curve) in advance by calculating the relationship between turbidity and microbial cell concentration, etc., based on the standard curve, The bacterial cell concentration and the like are determined from the measured turbidity of the culture solution.
  • Patent Document 1 Japanese Patent Laid-Open No. 2003-75344
  • Patent Document 2 Japanese Patent Application Laid-Open No. 2008-232790 proposes securing the length of a measurement optical path by providing a reflection mirror in a water quality measurement device that measures turbidity and the like.
  • a turbidity sensor is arranged in the culture tank by fixing it to the top plate (canopy) at the top of the culture tank. It is known to measure the turbidity of a culture solution.
  • the turbidity sensor when the turbidity sensor is placed in the culture tank, the turbidity sensor may be damaged by heat cycle due to autoclave sterilization of the entire small culture tank or steam sterilization in a stationary culture tank. If the turbidity sensor is damaged, it is practically impossible to replace it during the culture.
  • the present invention has been made in view of such problems of the conventional technology, and the object of the present invention is to accurately measure the turbidity of a culture solution in culturing microorganisms, animal and plant cells, etc. Therefore, it is intended to provide a technique capable of accurately obtaining the bacterial cell concentration and the like.
  • the present inventors have arranged a light emitting part and a light receiving part for detection on the outside of the tank wall toward the reflecting mirror arranged in the culture solution in the culture tank, Then, the culture medium is irradiated from the light emitting part through the transparent part provided on the tank wall, and the light from the reflector direction is received by the light receiving part for detection through the transparent part, thereby culturing from outside the culture tank.
  • the transmission distance (transmitted optical path length) of the light in the culture solution is at least 2 of the distance between the inner surface of the transparent part and the reflecting mirror. Since the absorption rate of light by the culture solution can be increased by extending it twice, the turbidity of the culture solution can be accurately measured using transmitted light even when the turbidity is low.
  • the light from the direction of the reflecting mirror received by the detection light-receiving unit includes scattered light as the turbidity increases, and finally becomes only scattered light.
  • the turbidity of the culture solution can be accurately determined by using scattered light even when the turbidity of the culture solution is high. It can be measured well.
  • the turbidity measuring apparatus includes a transparent portion provided on the tank wall of the culture tank, a reflector disposed in the culture solution in the culture tank, and disposed outside the culture tank.
  • a light emitting unit that irradiates light through the transparent unit, and a light receiving unit for detection that is disposed outside the culture tank and receives light from the direction of the reflecting mirror through the transparent unit.
  • another embodiment of the turbidity measuring apparatus further includes bubble removing means for preventing bubbles from being mixed in the turbidity measuring optical path between the inner surface of the transparent portion and the reflecting mirror.
  • the bubble removing means may be an optical path vicinity enclosure that surrounds at least a part of a space including the turbidity measuring optical path.
  • another embodiment of the turbidity measuring apparatus of the present invention is characterized in that the optical path vicinity enclosure is formed of a 10-200 mesh net.
  • the present invention is a culture apparatus, characterized in that it includes the turbidity measurement apparatus described above.
  • culture management of the culture solution can be performed quickly and accurately.
  • the turbidity of the culture solution can be monitored online, and thereby the bacterial cell concentration of the culture solution can be grasped and managed.
  • the present invention is a method for measuring the turbidity of a culture solution from outside the culture tank, and is characterized by using the turbidity measuring apparatus described above.
  • the present invention it is possible to optically measure the turbidity of a culture solution from outside the culture tank. Moreover, since the microbial cell density
  • the turbidity from the time point when the turbidity of the culture solution is low to the time when it is high can be continuously measured.
  • the reflecting mirror is arranged on the inner side of the culture tank, the light emitting part and the light receiving part for detection are arranged on the outer side of the reflecting tank, and the light emitting part is provided via the transparent part provided on the tank wall of the culture tank.
  • a turbidity sensor in the culture tank in order to receive light from the direction of the reflector, such as light transmitted through the culture medium between the transparent part inner surface and the reflector, Compared with the case where is placed, stable turbidity measurement can be performed without causing damage to the turbidity sensor due to thermal heat cycle.
  • a turbidity sensor it is not necessary to attach a turbidity sensor to the upper part of the culture tank as in the prior art, and this is particularly advantageous when measuring turbidity in a small culture tank in which it is difficult to secure a mounting space.
  • the turbidity of the culture solution can be measured optically from outside the culture tank.
  • the culture solution may or may not contain bubbles. In the latter case, the bubbles may cause an error in the measured turbidity.
  • turbidity can be measured without being greatly affected by bubbles.
  • FIG. 2 is a graph (standard curve) showing the relationship between turbidity (absorbance) measured in Example 1 and yeast cell concentration. It is a graph (standard curve) which shows the relationship between the turbidity (absorbance) measured in Example 2, and yeast cell density
  • FIG. 6 is a schematic cross-sectional view showing a tip portion of a coaxial optical fiber in a light emitting / receiving unit integrated light emitting / receiving unit used in Examples 3 to 5. It is a graph (standard curve) which shows the relationship between the turbidity (absorbance) measured in Example 3, and yeast cell density
  • the present invention relates to a turbidity measuring apparatus for optically measuring the turbidity of a culture solution from outside the culture tank.
  • turbidimeters there are four types of turbidimeters: a transmitted light measurement method, a scattered light measurement method, a transmitted light / scattered light calculation method, and an integrating sphere measurement method.
  • the turbidity is measured by using the transmitted light when the test solution containing light is irradiated, and in the scattered light measurement method, the turbidity is measured by using the scattered light from the test solution.
  • the basic configuration of the turbidity measuring apparatus of the present invention is that a reflecting mirror is arranged in a culture tank, a light emitting part and a light receiving part for detection are arranged outside the culture tank, and the light receiving part for detection is arranged on the tank wall of the culture tank.
  • Light is emitted from the light emitting part to the culture solution through the transparent part provided, and light from the reflector direction such as light transmitted through the culture solution between the inner surface of the transparent part and the reflector is detected on the same side as the light emitting part.
  • the light receiving unit receives light.
  • One feature of the present invention is that the turbidity of the culture solution is measured from the outside of the culture tank by a transmitted light measurement method by arranging reflected light in the culture tank.
  • the light emitting part and the light receiving part for detection are arranged on the same side outside the culture tank, that is, at substantially the same position or near the outside of the culture tank, as described later, by arranging the reflecting mirror in the culture tank.
  • the turbidity of the culture solution can be measured by a transmitted light measurement method.
  • the present invention even when the turbidity of the culture solution is low, the irradiation light from the light emitting part is reflected between the inner surface of the transparent part and the reflecting mirror. ), It is possible to measure the turbidity of the culture solution by the transmitted light measurement method by extending the transmitted light path length and increasing the absorption rate of the culture solution.
  • the other feature of the present invention is that, although the turbidity transmitted light measurement method is adopted, the light emitting unit and the light receiving unit for detection are arranged at substantially the same position outside the culture vessel or in the vicinity thereof. In addition, since the light emitting unit and the light receiving unit for detection are both arranged toward the reflecting mirror, the light receiving unit for detection is incident in the direction of the reflecting mirror in the culture solution of light caused by irradiation light. It is also possible to receive scattered light from turbid substances (bacteria, cells, etc.).
  • the turbidity measuring apparatus of the present invention is not limited to the transmitted light measurement method, but is used in a wide range from low to high turbidity as a transmitted light / scattered light measurement method and further as a scattered light measurement method.
  • the turbidity of the culture solution can also be continuously measured, and it is extremely effective for the temporal management of the cell concentration and the like such as grasping the growth process in the culture of microorganisms, animals and plants.
  • the turbidity measuring apparatus of the present invention is provided with a transparent portion provided on the tank wall of the culture tank, a reflecting mirror disposed in the culture solution in the culture tank, and disposed outside the culture tank and reflecting.
  • a light emitting unit that emits light toward the mirror through the transparent unit, and a light receiving unit for detection that is disposed outside the culture tank and receives light from the direction of the reflecting mirror through the transparent unit.
  • the culture tank is used by putting a culture solution therein, and a known culture tank can be used.
  • the material, size, shape and the like of the culture tank are not particularly limited, and can be appropriately selected in consideration of the application, culture / sterilization temperature, and the like.
  • the material of the culture tank is preferably, for example, metal, glass, or resin. From the viewpoint of transparency and heat resistance in the small culture tank, for example, transparent heat-resistant glass such as Pyrex (registered trademark) glass, stainless steel in the large culture tank. Steel or the like is preferred.
  • the volume of the culture tank is not particularly limited, and any capacity culture tank can be used.
  • the culture tank can be used from a small culture tank having a capacity of about 100 ml to a large culture tank having a capacity exceeding 10,000 L.
  • the shape of the culture tank may be, for example, a cylindrical shape, a conical shape, a prismatic shape, or a pyramid shape.
  • the opening part of a culture tank may be liquid-tightly sealed with a top plate etc. so that it may mention later.
  • the tank wall of a culture tank means the wall surface which comprises a culture tank, and contains not only the side wall of a culture tank but a bottom wall.
  • the transparent portion is provided on the side wall or bottom wall of the culture tank because it is easy to measure the turbidity of the culture solution in contact with the inner surface thereof.
  • the transparent part is provided on the tank wall of the culture tank as described above, and transmits the irradiation light, transmitted light and scattered light when measuring turbidity.
  • the tank wall of the culture tank is transparent
  • the tank wall can be used as a transparent part, but when the tank wall of the culture tank is made of an opaque material such as metal, a transparent material is partially used. It is only necessary to provide a transparent portion by using.
  • the material of the transparent part is preferably heat resistant from the viewpoint of the sterilization treatment of the culture tank.
  • glass, resin or the like can be used.
  • the transparent resin include polypropylene (PP) and polycarbonate (PC). And tetrafluoroethylene / hexafluoropropylene copolymer (FEP).
  • the light emitting section and the detection light receiving section are provided outside the transparent section.
  • the turbidity measuring apparatus of the present invention achieves excellent durability and measurement accuracy.
  • the outer side of the transparent part refers to the side facing the transparent part through the transparent part when the side in which the culture solution is stored is taken as the inner side.
  • a reflector is arranged in the culture tank, and by adopting such a configuration, as described above, the turbidity measuring apparatus of the present invention has excellent measurement accuracy even when turbidity is low. Realize. In other words, by increasing the light transmission distance by reciprocating the culture solution between the transparent part and the reflector through a reflector, the light absorption rate by the turbid substance in the culture solution is increased. Therefore, even when the turbidity of the culture solution is low, the bacterial cell concentration and the like can be optically measured as turbidity from outside the culture tank with high accuracy.
  • the installation position of the reflecting mirror is not particularly limited, and is installed at an arbitrary position in the culture solution having no light shielding material such as equipment and members in the optical path of the transmitted light between the transparent portion and the reflecting mirror. .
  • the position is selected in consideration of the position of irradiation light so that the amount of received light or the received light value (hereinafter referred to as “received light amount or the like”) in the detection light receiving unit is maximized.
  • the reflecting mirror is set, for example, at a position of 1 to 50 mm, preferably 2 to 30 mm from the inner surface of the transparent portion of the tank wall.
  • a light emitting part is arranged outside the side wall of the culture tank, and a light receiving part for detection is arranged outside the opposite side wall to measure turbidity by a transmitted light measurement method. It is difficult to do because there are light shields such as various devices and members in the culture tank, but by using the turbidity measuring device of the present invention, the light from the light emitting part outside the tank wall is reflected.
  • the turbidity can be measured from the outside of the culture tank by the transmitted light measurement method by reflecting the light with a mirror and receiving the light with the detection light receiving unit outside the same side.
  • the angle of the reflection surface is not particularly limited as long as the transmitted light can be measured, but the reflection surface faces the inner surface of the transparent portion, and the irradiation light from the light emitting portion is used as the detection light receiving portion. It is preferable that they are arranged at an angle that allows efficient reflection.
  • the reflector may be supported by a device or member such as a baffle plate installed in the culture tank, or a dedicated support member can be used. And measurement errors due to position movement can be prevented.
  • the material of the reflecting mirror is not particularly limited, but for example, metal, glass, and resin are preferable. From the viewpoint of sterilization, stainless steel and heat resistant glass are more preferable, and the reflecting surface is coated with a metal such as aluminum or silver. It may be a thing.
  • the shape of the reflecting mirror is not particularly limited as long as it reflects light, and reflecting mirrors of any shape such as a plane mirror, a concave mirror, and a convex mirror can be used. Further, the size and shape of the reflecting surface of the reflecting mirror are not particularly limited. For example, a circular mirror or a rectangular mirror having a diameter of 10 to 30 mm can be used.
  • the reflector placed in the culture tank can be separately installed in the culture tank, but the surface of various devices and members such as stainless steel sensors in the culture tank is reflected as it is or after being processed. Can be used as a mirror.
  • the surface of an object in the culture tank since the space in the culture tank is limited, it is advantageous to use the surface of an object in the culture tank as a reflecting mirror.
  • the number of reflectors arranged in the culture tank is not limited to a single one, and a plurality of reflectors may be arranged at positions where the distance from the inner surface of the transparent part of the tank wall is different and the turbidity measurement optical paths do not overlap.
  • a plurality of reflectors may be arranged at positions where the distance from the inner surface of the transparent part of the tank wall is different and the turbidity measurement optical paths do not overlap.
  • the turbidity of the culture solution is low, it is difficult to measure turbidity when using a certain reflector, but turbidity can be measured using another reflector that has a longer distance from the inner surface of the transparent part.
  • the light emitting / receiving units described later disposed outside the culture tank may be moved to positions facing the reflecting mirrors when measuring turbidity.
  • a belt-like reflecting mirror separated from the inner surface of the transparent part of the side wall at an appropriate distance all around (the reflecting mirror at this time should be cylindrical) can be arranged within an appropriate range such as a half circle.
  • the light emitting / receiving unit described later arranged outside the culture tank is restricted in the vertical direction but can be arranged with a degree of freedom in the horizontal direction.
  • the portion of the side wall facing the reflecting mirror at this time is a transparent portion.
  • a light emitting / receiving unit described later is disposed below the transparent window of the supporting rigid flat plate on which the entire bottom of the transparent wide bag is fixed and placed, and the upper surface of the transparent window
  • the turbidity of the present invention is such that a reflecting mirror is fixed inside a tank wall (transparent portion of the bottom wall) in contact with the bag through a support member fixed to the inner surface of the bag with an adhesive or the like.
  • a measuring device can be applied.
  • the irradiation light can be generated using a light source usually used for measuring turbidity.
  • the wavelength of light is not particularly limited, for example, light having a wavelength of 900 nm, 660 nm, 600 nm, 610 nm, 562 nm, or the like can be suitably used.
  • a publicly known one can be used as the light emitting part of the irradiation light.
  • a light emitting diode LED: Light Emitting Diode
  • a laser can be suitably used as the light source.
  • the turbidity measuring apparatus of the present invention is provided with a light receiving part for detection.
  • the light receiving unit for detection is a position that can receive light from the direction of the reflecting mirror caused by irradiation light such as light reflected by the reflecting mirror and transmitted through the culture solution, and facing the reflecting mirror outside the transparent portion. Be placed.
  • a publicly known one can be used.
  • a photodiode Photo-Diode: PD
  • a photocell or the like can be preferably used.
  • each of the irradiation and light receiving parts facing the outer surface of the transparent part of the tank wall may be a separate type or an integrated type.
  • the light emitting unit (light projector) and the light receiving unit for detection (light receiver) are separate, and these are connected to the arithmetic processing unit (amplifier) attached to the digital display unit via a cable. Can be linked.
  • the light emitting unit includes a main body that houses the light source and an optical fiber connected to the main body, and light is irradiated from the tip of the optical fiber.
  • the detection light receiving section includes a main body for housing the PD and an optical fiber, and the light received at the tip of the optical fiber is detected by the PD.
  • the main body of the light emitting unit and the main body of the light receiving unit for detection may be combined with an arithmetic processing unit (amplifier) attached to the digital display unit to form one unit.
  • the portion for irradiation and light reception at this time may be a separate type or an integrated type.
  • the tip of one optical fiber connected to the light source in the light emitting unit body May be a coaxial one in which the tip ends of a plurality of optical fibers connected to the PD in the detection light receiving unit main body are arranged around the center axis.
  • bubble removing means is provided inside the transparent portion. Furthermore, it can be provided.
  • an optical path vicinity enclosure that surrounds at least a part of the space including the turbidity measuring optical path can be cited.
  • a peripheral surface substantially perpendicular to the inner surface of the transparent portion that is, , Side surface
  • the optical path vicinity enclosure has one end surface in contact with the inner surface of the transparent portion of the tank wall, the contact surface opened, and the other end surface formed of a reflecting mirror.
  • the entire circumference of the space including the turbidity measurement optical path (in other words, the angle of the circumferential surface in the direction substantially perpendicular to the inner surface of the transparent portion is 360 degrees).
  • the surrounding optical path surrounding body can be exemplified.
  • an optical path vicinity enclosure which is surrounded by a material (for example, a net) that allows the culture medium to pass but does not allow air bubbles to substantially pass and has a non-contact surface with the tank wall can be exemplified.
  • substantially do not allow bubbles to pass means that fine bubbles may be allowed to pass therethrough.
  • bubbles that may be generated by the collection of fine bubbles that have passed through can be naturally discharged to the outside of the enclosure near the optical path by the buoyancy. It is convenient because it can.
  • the optical path vicinity enclosure is an optical path vicinity enclosure having an open portion in a direction substantially perpendicular to the inner surface of the transparent part, and at least a half circumference of a peripheral surface of the space including the turbidity measurement optical path (
  • it may be one that is surrounded by a member that does not allow the culture medium to pass through, and that removes bubbles by retaining the culture liquid.
  • the turbidity of the inside and outside of the enclosure near the optical path is made uniform by the inflowing culture solution staying, but the inflowed bubbles are naturally discharged from the open part of the peripheral surface part to the outside of the enclosure near the optical path. be able to.
  • the space including the turbidity measurement optical path has a shape surrounding an angle of 180 degrees or more and less than 360 degrees of the circumferential surface substantially perpendicular to the inner surface of the transparent portion (in the same direction). Any shape can be used as long as it has a belt-like open portion on the peripheral surface), but a substantially L-shaped or substantially U-shaped cross section is preferable.
  • turbidity is an index indicating the degree of turbidity of water.
  • a standard prepared by using a standard solution or the like which is a known method by measuring the amount of transmitted light or scattered light received. From the curve (calibration curve), the concentration of the item required for the sample can be quantified as turbidity.
  • the absorbance, the received light value that is the received light value, etc. can be measured as turbidity.
  • a standard curve showing the relationship between turbidity and cell concentration (cells / ml) is a known method, for example, a turbidity measurement value of a culture solution and a known method for the culture solution at that time. Or from the cell suspensions prepared to various known cell concentrations and the respective turbidity measurement values. Since this standard curve differs depending on the type of microorganism, animal or plant cell, etc., it is prepared in advance for each type, and the effective measurement range such as the bacterial cell concentration can also be confirmed from this standard curve.
  • the turbidity of a culture solution to be measured is such that the absorbance measured using a spectrophotometer (for example, absorbance at a wavelength of 660 nm) is in the range of 0.2-6. Sometimes, it is applied particularly effectively, but is not limited to such a range.
  • the turbidity measuring apparatus of the present invention can be measured from the outside, it can be applied regardless of the internal environment. Therefore, small-scale basic research and large-scale industrial use can be applied to all cultures such as microorganisms and cells. Can be used for various purposes.
  • a culture apparatus provided with the turbidity measuring apparatus of the present invention and a method for optically measuring the turbidity of a culture solution from the outside of the culture tank using the turbidity measuring apparatus of the present invention. Can be mentioned.
  • FIG. 1 is a schematic view showing an embodiment of the turbidity measuring apparatus of the present invention
  • FIG. 1 (a) is an explanatory schematic view showing a part thereof in a longitudinal section
  • FIG. It is explanatory explanatory drawing which shows the cross section by X.
  • the culture apparatus 1 is a culture apparatus provided with a stirrer 3 and a baffle plate (baffle plate) 4 in a cylindrical culture tank 2, and is a turbidity measurement apparatus 6A according to an embodiment of the present invention.
  • the culture tank 2 illustrated in FIG. 1 is made of heat-resistant glass, which is Pyrex (registered trademark) glass, and has a capacity of 1000 ml.
  • sensors and the like provided as necessary in the culture tank 2 such as a pH sensor, a DO sensor, a vent pipe, a temperature control pipe, an exhaust port, and a temperature control pipe are shown. However, it is natural that these can be attached to the culture tank of FIG. 1 according to the embodiment.
  • the upper opening of the cylindrical culture tank 2 is liquid-tightly sealed with a top plate 5.
  • the culture tank 2 is equipped with a stirrer 3, and for example, the rotating shaft 3a is supported by a bearing 3b provided at the center on the top plate 5, and the stirring blade 3c is attached to the rotating shaft 3a in multiple stages. It has been.
  • a rotating disk 3d containing a plurality of permanent magnets (not shown) is connected to the lower end of the rotating shaft 3a so as to be separated from the inner surface of the bottom wall 2b, and the rotating disk 3d is separated from the outer surface of the bottom wall 2b. It is rotated by a magnet rotating body (not shown) that faces the lever and rotates horizontally by power.
  • baffle plates 4 are suspended in the axial direction in the vicinity of the inner periphery of the culture tank 2 (only one is shown in FIG. 1a), and each upper end of the baffle plate 4 is suspended and fixed to the top plate 5. Further, the lower end portions thereof are firmly connected and fixed to each other by a ring 4a (a part of which is shown in FIG. 1A).
  • a light emitting / receiving unit (for example, using laser light) 6U is arranged outside the tank wall of the culture tank 2.
  • Separate light-emitting portions 6a (example: laser projector) and detection light-receiving portions 6b (example: light receiver using PD) are respectively fixed as light-receiving / emitting units 6U and housed in the casing.
  • the reflecting mirror 8 (example: via the transparent part 2aa).
  • a light emitting section 6a that irradiates light toward a stainless steel circular mirror having a diameter of 20 mm is disposed, and the light irradiated from the light emitting section 6a such as light reflected by the reflecting mirror 8 and transmitted through the culture solution 7 is disposed.
  • a light receiving unit 6b for detection that receives light from the direction of the reflecting mirror through the transparent part 2aa and outputs a light reception signal for detection according to the amount of received light or the like is arranged toward the reflecting mirror 8.
  • the arrangement position of the light emitting / receiving unit 6U may be any as long as the culture solution 7 is outside the tank wall in contact with the inner surface thereof.
  • the position of the reflecting mirror 8 (example: substantially perpendicular direction from the inner surface of the transparent portion 2aa to the inner surface thereof. 3mm) is appropriately selected and determined.
  • the light emitting / receiving unit 6U may be installed on the outside of the tank wall by an appropriate method.
  • the light receiving / emitting unit 6U may be directly attached to the tank wall with a support member whose distance from the tank wall can be adjusted, or may be mounted on a support stand near the tank wall. It may be attached or the like.
  • the light emitting / receiving unit 6U may accommodate an arithmetic processing unit attached to the digital display unit for digitizing the light receiving amount or the like, or for calculating turbidity.
  • a position where the amount of received light is maximized is generally selected as the installation position of the reflector 8 that is fixedly arranged in the culture tank 2.
  • the reflection surface faces the inner surface of the transparent portion 2aa and is disposed in parallel with the axial direction.
  • the reflecting mirror 8 in the turbidity measuring apparatus 6A shown in FIG. 1 is provided inside the transparent part 2aa of the side wall 2a. However, when this is provided in the transparent part of the bottom wall 2b, the reflecting mirror 8 is provided on the side wall 2a. It may be provided in accordance with the same purpose as provided.
  • the turbidity measuring apparatus 6A of the present invention configured as described above allows the irradiation light from the light emitting part 6a to pass through the transparent part 2aa through the reflector 8 even when the turbidity of the culture solution 7 is low. Transmission through the culture solution 7 from the inner surface to the reflection mirror 8 and further from the reflection mirror 8 to the inner surface of the transparent portion 2aa in the direction of the detection light receiving portion 6b, that is, the transmission distance (transmission of light in the culture solution 7) By increasing the optical path length), the absorption rate of light by suspended substances (turbid substances) in the culture solution is increased, and the cell concentration etc. is optically measured as turbidity from outside the culture tank 2 with high accuracy. can do.
  • the culture apparatus 1 measuring the turbidity by the transmitted light measurement method by arranging the light emitting part outside the tank wall and arranging the light receiving part for detection outside the opposite tank wall is the culture tank 2.
  • the culture tank 2 Inside, it is difficult because there are light shielding materials such as various devices and members as described above, but in the turbidity measuring device 6A of the present invention, the light from the light emitting portion 6a outside the tank wall is reflected.
  • the turbidity can be measured mainly from the outside of the culture vessel 2 by the transmitted light measurement method by reflecting the light by the mirror 8 and receiving the light by the detection light receiving unit 6b outside the same side.
  • the reflecting mirror 8 is supported by a baffle plate 4 made of stainless steel that is suspended in the axial direction of the culture tank 2.
  • a notch concave portion
  • the reflecting mirror 8 is firmly fitted thereto, screwed via an attachment member
  • an appropriate method such as welding is adopted.
  • apparatuses and members other than the baffle plate 4 can also be utilized suitably, and a dedicated support member can also be used.
  • the reflecting mirror 8 can be firmly fixed to the baffle plate 4 or the like, thereby preventing measurement errors due to vibration or position movement of the reflecting mirror.
  • the reflecting mirror 8 As a method of arranging the reflecting mirror 8 at a predetermined position in the culture tank 2, the reflecting mirror 8 is previously placed outside the culture tank 2 at a specific position of the baffle plate 4 (when the baffle plate 4 is inserted into the culture tank 2) 8 is a predetermined position), and then the top plate 5 to which the upper end of the baffle plate 4 is fixed is gradually lowered to insert the baffle plate 4 into the culture tank 2, and then the top plate 5 Is placed on the upper opening surface of the culture tank 2 and fixed so that the reflecting mirror 8 is appropriately disposed at a predetermined position in the culture tank 2.
  • the positional relationship between the light emitting unit 6a and the detection light receiving unit 6b in the turbidity measuring device 6A is not particularly limited, and may be any direction such as a vertical direction or a horizontal direction as in the present embodiment.
  • the light emitting unit 6a and the detection light receiving unit 6b are positions where the optical axis of the irradiation light from the light emitting unit 6a and the optical axis of the incident light (transmitted light) to the detection light receiving unit 6b are necessarily included in the same plane. In addition, it is usually arranged at a position where the angle of both optical axes is an appropriate angle of 100 degrees or less, for example, a position where the amount of received light is maximized.
  • the detection light-receiving unit 6b also receives external light other than the light emitted from the light-emitting unit 6a.
  • the intensity of light as an external light source of the light-emitting unit 6a is external.
  • the received external light signal is processed by a filter circuit to obtain an electric signal (target,
  • the turbidity can be measured without being affected by external light by adopting a method such as analog circuit processing of only the received light light signal from the pulsed light source. For this reason, light shielding in the vicinity of the optical path between the light emitting part 6a and the detection light receiving part 6b from the outer surface of the transparent part 2aa of the tank wall is not particularly required.
  • the light emitting unit 6a may be a single light source or a plurality of light sources, and the light source light intensity and the like are not particularly limited and are appropriately selected.
  • the detection light-receiving unit 6b is disposed toward the reflecting mirror 8 and receives light from the direction of the reflecting mirror 8 due to irradiation light such as transmitted light from the reflecting mirror 8. Accordingly, the amount of light received by the detection PD (detection light receiving portion 6b) (detection light reception signal) varies depending on the turbidity of the culture solution 7 between the inner surface of the transparent portion 2aa and the reflecting mirror 8.
  • the detection PD outputs a detection light reception signal (electrical signal) that changes according to the turbidity, and the arithmetic processing unit (not shown) quantifies the electric signal, and further, based on this, the turbidity is calculated. It is calculated by a method well known to those skilled in the art.
  • the light emitting / receiving unit 6U having the light emitting unit 6a and the detection light receiving unit 6b for example, as described above, using laser light
  • a known unit can be used, for example, a transmission type laser sensor head IB. -01 (manufactured by Keyence Corporation) is used.
  • This light emitting / receiving unit is connected to a known arithmetic processing unit attached to the digital display unit, for example, a transmission type laser sensor amplifier 1B-1500 (manufactured by Keyence Corporation) via a cable.
  • a light emitting / receiving unit in the case where an LED or the like is used as a light source is as exemplified in the examples described later.
  • the turbidity measuring apparatus 6A of the present invention thus configured can accurately measure the turbidity of the culture solution 7 from the outside of the culture tank 2, and the measured turbidity and the turbidity in advance From the standard curve for which the relationship with the bacterial cell concentration or the like is required, the bacterial cell concentration or the like of the culture solution is determined.
  • the turbidity measurement may be performed every predetermined time or continuously.
  • the turbidity measuring apparatus of the present invention can include a bubble removing means.
  • FIG. 2A is a longitudinal sectional view showing an embodiment of the turbidity measuring apparatus of the present invention provided with bubble removing means
  • FIG. 2B is a schematic view showing a section taken along YY. 3 (a) and 3 (b) are enlarged schematic views of the turbidity measuring device portion of FIG.
  • members and places that are substantially the same as those in the above embodiment are denoted by the same reference numerals, and description thereof is omitted.
  • the bubble removing means substantially prevents bubbles generated by aeration stirring or the like in the culture solution in the turbidity measuring optical path between the inner surface of the transparent part and the reflecting mirror, and may be generated by bubbles in some cases. Suppress errors in certain turbidity measurements.
  • the turbidity measuring apparatus of the present invention provided with the bubble removing means can be used even in the case of turbidity measurement of a culture solution containing no bubbles (bubbles are not substantially mixed).
  • the bubble removing means 6c provided in the turbidity measuring device 6B includes a turbidity measuring optical path between the inner surface of the transparent portion 2aa and the reflecting mirror 8.
  • the entire circumference in other words, the portion corresponding to the circumferential surface of the space is indicated by 6cs in FIG. 3B) in the direction substantially perpendicular to the inner surface of the transparent portion 2aa (in other words, the circumferential surface 6cs in the same direction).
  • the optical path vicinity enclosure (6c) in which the angle of 360 degrees is enclosed, and is disposed sideways so as to sufficiently surround the turbidity measurement optical path.
  • the bubble removing means 6c is, for example, a cylindrical (or rectangular tube-shaped) optical path vicinity enclosure (6c), and the optical path vicinity enclosure (6c) is a contact surface (one side) with the inner surface of the transparent portion 2aa.
  • the transparent portion 2aa is formed by the reflecting mirror 8, and the other surface (portion) in contact with the culture solution 7 passes the turbid substance but the air bubbles substantially pass.
  • the net 6ca is not formed.
  • what is necessary is just to adhere
  • the size of the enclosure near the optical path (6c) is not limited, but the inner diameter is preferably 10 to 30 mm, and the height is preferably 2 to 30 mm.
  • the inner diameter is the diameter of the reflector 8 and the height is transparent.
  • the distance from the inner surface of the portion 2aa to the reflecting mirror is preferably the same.
  • a part of the enclosure near the optical path may function as the reflecting mirror 8, and in this case, the opposing surface of the transparent portion 2 a forms the reflecting mirror 8.
  • bubble removing means 6c having such a simple structure, bubbles mixed in the culture solution 7 in the vicinity of the turbidity measuring optical path can be substantially removed, and the object of the present invention can be achieved.
  • a bubble discharge port 6cb is provided in the upper part of the optical path vicinity enclosure (6c), and the opening area can be 100% of the upper surface or the other end surface, but 20 to 80% is preferable.
  • the upper part means the upper surface (above half of the cross section of the cylinder) or the upper part of the other end surface, and this also applies to the following.
  • the bubble discharge port (opening) 6cb is provided so that the fine bubbles may be formed by gradually gathering the bubbles. By releasing from the air, it is possible to prevent the bubbles from affecting the turbidity measurement value.
  • a skeleton is formed by an appropriate support member, and a surface in contact with the culture solution 7 is formed by a net 6ca along the skeleton.
  • a material of the support member for example, a metal, a heat resistant resin, or the like is used from the viewpoint of sterilization treatment of the culture tank 2.
  • the material of the mesh 6ca is not particularly limited, and similarly, for example, metal, heat-resistant resin, etc. can be mentioned, and the mesh is, for example, 10 to 200 mesh, preferably 20 to 100 mesh.
  • the optical path vicinity enclosure (6c) having the reflecting mirror 8 on the other end face is disposed, for example, so that its central axis is substantially perpendicular to the inner surface of the transparent portion 2aa of the side wall 2a.
  • the reflecting mirror 8 is supported by the baffle plate 4 suspended in the axial direction of the culture tank 2, the outside of the reflecting mirror 8 on the other end face is bubbled.
  • the entire enclosure near the optical path (6c) may be supported by supporting the baffle plate 4 with the discharge port 6cb facing up.
  • the contact surface between the opening end surface of one end surface of the enclosure near the optical path (6c) and the inner surface of the transparent portion 2aa may be brought into close contact via an elastic body (not shown) or the like.
  • the material of the elastic body is not particularly limited, and for example, silicon resin can be used.
  • an elastic member such as a spring is interposed between the other end face of the optical path vicinity enclosure (6c) and the baffle plate 4, and the opening end face of one end face of the optical path vicinity enclosure (6c) is transparent.
  • the contact surface with the inner surface of the portion 2aa may be pressed in close contact with the transparent portion 2aa.
  • An example of a method of mounting the enclosure near the optical path (6c) in the culture tank 2 is substantially the same as the case where the reflecting mirror 8 is disposed, and the elastic body is provided on the opening end surface of one end surface as necessary.
  • the fixed top plate 5 is gradually lowered and the baffle plate 4 is inserted into the culture tank 2, and then the top plate 5 is placed and fixed on the upper end opening surface of the culture tank 2 to thereby enclose the vicinity of the optical path.
  • (6c) is appropriately arranged in the vicinity of the optical path in the culture tank 2.
  • the optical path vicinity enclosure (6c) arranged and mounted in this way has an opening end surface of one end surface thereof in close contact with the inner surface of the transparent portion 2aa, and the other end surface thereof is firmly fixed to the baffle plate 4. It does not vibrate or move position.
  • the bubble removing means 6c shown in FIGS. 2 and 3 is provided in the transparent portion 2aa of the side wall 2a.
  • the above-described reflecting mirror 8 is disposed.
  • the enclosure near the optical path is substantially perpendicular to the inner surface of the transparent portion of the bottom wall 2b, and the bubble outlet is in the vicinity of the optical path. What is necessary is just to provide in the other end part surface (upper part) of an enclosure.
  • the turbidity measuring apparatus 6B of the present invention including the bubble removing means 6c configured as described above eliminates or reduces errors in turbidity measurement values that may be caused by bubbles in the culture solution 7, and more accurately. Turbidity can be measured.
  • the enclosure near the optical path is at least a half circumference (angle 180) of the circumferential surface substantially perpendicular to the inner surface of the transparent portion of the space including the turbidity measuring optical path. Is formed of a material that does not allow the culture medium to pass therethrough, and thereby the culture medium is retained to remove bubbles in the culture liquid.
  • the turbidity is an enclosure near the optical path having a substantially L-shaped or substantially U-shaped cross section, with one end face opening to contact the inner surface of the transparent portion of the tank wall.
  • a bubble removing means that surrounds the measurement optical path with sufficient spread can be mentioned.
  • the other end faces are provided with reflecting mirrors.
  • the bubble of the culture fluid flows in a substantially constant direction by the stirring method, the aeration method, etc., or flows as a turbulent flow, but what type of optical path vicinity enclosure is used, Taking into account the flow state of the bubbles, such as using a near-L-shaped optical path enclosure with a substantially L-shaped cross section when the flow of bubbles is in a substantially constant direction, and a near-U-shaped optical path enclosure with turbulent flow. What is necessary is just to select suitably.
  • FIG. 4 is a schematic perspective view showing a bubble removing means which is an optical path vicinity enclosure having an L-shaped cross section, and has a very simple structure.
  • the one end face 16cc is in contact with the inner surface of the transparent portion of the tank wall, and the outside of the L-shaped side face 16cd is located upstream of the flow of the culture medium bubbles.
  • a retention area of the culture solution is formed between the inside of one peripheral surface (side surface) 16cd and the inside of the other L-shaped peripheral surface (side surface) 16ce.
  • Bubbles are removed from 16cf.
  • at least half the circumference (angle 180 degrees) of the circumferential surface substantially perpendicular to the inner surface of the transparent portion is surrounded by the inner side of 16cd and 16ce.
  • the bubble removing means 16c if the outside of the other side surface 16ce of the L-shape is arranged so as to be positioned on the lower peripheral surface (side surface) of the turbidity measuring optical path, the bubble rising from below is provided. This is preferable because it can prevent the contamination.
  • One end face 16cc of the bubble removing means 16c is in contact with the inner surface of the transparent portion of the tank wall, but a reflecting mirror 18 is provided on the other end face.
  • FIG. 5 is a schematic perspective view showing a bubble removing means which is an optical path vicinity enclosure having a U-shaped cross section and a cross-sectionally arc-shaped cross section. In these cases, most of the peripheral surface (side surface) near the turbidity measurement optical path is surrounded by the optical path vicinity enclosure.
  • one end face 26cc, 36cc is in contact with the inner surface of the transparent portion of the tank wall, but the other end face is provided with reflecting mirrors 28, 38. And since the residence area
  • the open portions 26cf and 36cf in a direction substantially perpendicular to the inner surface of the transparent portion on the peripheral surface (side surface) of the optical path vicinity enclosures (26c) and (36c) are positions where air bubbles are easily released as much as possible (normally upward). It is preferable to set to.
  • a turbidity measurement optical path vicinity between the transparent part and the reflecting mirror can be sufficiently surrounded, for example, a cylindrical optical path vicinity enclosure or the like.
  • the other end face can be made to function as a reflecting mirror by using a material that can be used for the reflecting mirror.
  • the enclosure near the optical path is, for example, cylindrical, and has a contact surface with the inner surface of the transparent portion at one end surface thereof, and has a culture solution outflow inlet at the top.
  • the inflowing culture solution stays and bubbles rise upward, and are discharged and removed from the upper culture solution outlet.
  • the material for the enclosure near the optical path used for removing bubbles by retaining the culture solution for example, metal, heat-resistant resin, rubber, or the like is used as the material for the enclosure near the optical path used for removing bubbles by retaining the culture solution.
  • the arrangement of the enclosure near the optical path in the culture tank, the supporting method, and the like may be the same as described in the above embodiment (FIGS. 2 and 3).
  • an optical path vicinity enclosure having a peripheral surface (side surface) perpendicular to the inner surface of the transparent portion of the tank wall is illustrated, but if it surrounds the vicinity of the turbidity measurement optical path, the angle of the surface Is not limited, and may be, for example, a tapered shape from the inner surface of the transparent portion of the tank wall toward the reflecting mirror.
  • the light emitting / receiving unit of the turbidity measuring apparatus outside the culture tank is temporarily removed. Etc., and move the culture tank, return to the original position after sterilization, and re-install the light emitting unit etc.
  • enclose the vicinity of the optical path with the light receiving and emitting unit etc. and the reflector in the culture tank The position with the body may be greatly displaced (for example, in the turbidity measuring apparatus shown in FIG. 2, the displacement in the horizontal direction). Taking this into consideration, the horizontal direction of the vertical cross section with respect to the axial direction of the enclosure near the optical path can be made sufficiently wide.
  • the turbidity measuring apparatus of the present invention provided with the bubble removing means configured as described above eliminates or reduces the error of the turbidity measurement value that may be caused by bubbles in the culture solution in some cases, and more accurately turbidity. Can be measured.
  • Example 1 (light emitting part: laser pulse light source)
  • the turbidity of a yeast suspension (culture solution) having a known yeast cell concentration was measured using the turbidity measuring apparatus 6A of the present invention using a laser pulse light source as a light emitting unit provided in the culture apparatus shown in FIG.
  • turbidity measuring device On the outside of a 1000 ml cylindrical culture tank (inner diameter 84 mm, height 180 mm) made of transparent glass, equipped with a light emitting / receiving unit in the turbidity measuring apparatus of the present invention (as exemplified in the turbidity measuring apparatus in FIG. 1), A culture apparatus equipped with a reflector in the culture tank was used. In this case, the glass on the side wall of the cylindrical culture tank constitutes the transparent part of the turbidity measuring apparatus of the present invention.
  • the light receiving / emitting unit is a separate light emitting / receiving unit (stock) with a laser pulse light source (wavelength: 660 nm) as a light emitting part (projector) and a photodiode (PD) as a light receiving part for detection (light receiver).
  • a transmission type laser sensor head IB-01 manufactured by Keyence Corporation was used.
  • the light emitting / receiving unit is installed so that the light emitting part is positioned 15 mm from the level of the bottom surface of the culture tank (the interval between the light emitting part and the light receiving part for detection is 30 mm). It was connected to a KEYENCE transmission laser sensor amplifier IB-1500.
  • the arithmetic processing unit includes a digital display unit, and the transmittance (%) is displayed on the digital display unit.
  • the combined device of the light emitting / receiving unit and the arithmetic processing unit is not affected by external light other than the irradiation light from the light emitting unit.
  • the light emitting unit and the detection light receiving unit of the light receiving and emitting unit are arranged at a position where the optical axis of the irradiation light from the light emitting unit and the optical axis of the incident light (transmitted light) to the detection light receiving unit are necessarily included in the same plane.
  • both optical axes are arranged so that the angle is 90 degrees (the angle of the optical axis of the irradiated light with respect to the reflecting surface of the reflecting mirror is 45 degrees and the angle of the optical axis of the incident light is 45 degrees).
  • the plane including the optical axis of the irradiation light and the optical axis of the incident light is perpendicular to the bottom surface of the culture tank, and the irradiation light is irradiated from below as shown in FIG. Received light.
  • the reflecting mirror is made of a circular (diameter 20 mm) stainless steel, and one of the three baffle plates is arranged so that its reflecting surface is opposed to and parallel to the inner surface of the transparent part of the side wall of the culture tank.
  • the reflecting mirror has a distance from the inner surface of the transparent portion of the tank wall to the reflecting surface of the reflecting mirror in the vertical direction (hereinafter referred to as “distance from the inner surface of the transparent portion to the reflecting mirror”) of 3.5 mm or 5. It arrange
  • the turbidity measuring optical path length (hereinafter simply referred to as “optical path length”) is 10 mm, and the distance from the inner surface of the transparent part to the reflecting mirror is 5.0 mm. In this case, the optical path length is 14 mm.
  • the light emitting and receiving unit with the light emitting part and the light receiving part for detection fixed to the outside of the culture tank so that the separation distance from the tank wall can be adjusted, and the separation distance can be adjusted so as to maximize the amount of light received. It is.
  • Saccharomyces cerevisiae Oriental Yeast Industry Co., Ltd., Oriental East Regular was used and suspended in water. was prepared.
  • Example 2 (light emitting part: LED pulse light source, light emitting part / light receiving part separate type) A yeast suspension having a known yeast cell concentration in the same manner as in Example 1 except that the following light emitting / receiving unit and arithmetic processing unit were used and the distance from the inner surface of the transparent part to the reflecting mirror was changed to the following distance. The turbidity of was measured.
  • Light emitting / receiving unit and arithmetic processing unit As a light receiving / emitting unit, a light emitting unit composed of an LED pulse light source (wavelength 660 nm) and an optical fiber coupled thereto, and a light receiving unit for detection composed of a PD and an optical fiber coupled thereto A separate light emitting / receiving unit (manufactured by Keyence Corporation, fiber unit FU-77 (fiber strand diameter 1.13 mm, with a transmissive lens F-4 attached to the tip of the fiber unit)) was used. This light emitting / receiving unit is connected to an arithmetic processing unit (Keyence Co., Ltd., digital fiber amplifier FS-N11MN) attached to the digital display unit to form one unit.
  • arithmetic processing unit Keyence Co., Ltd., digital fiber amplifier FS-N11MN
  • Example 3 (light emitting part: LED pulse light source, light emitting part / light receiving part integrated type) Using the following light emitting / receiving unit and arithmetic processing unit, except that the distance from the inner surface of the transparent part to the reflecting mirror is 7.5 mm (optical path length: 15 mm), and the yeast cell concentration is as described in Table 3, The turbidity of a yeast suspension with a known yeast cell concentration was measured in the same manner as in Example 2.
  • Light emitting / receiving unit and arithmetic processing unit a light emitting / receiving unit integrated type (coaxial type) light receiving / emitting unit was used. This light emitting / receiving unit is composed of an optical fiber unit, a light emitting part, and a light receiving part for detection.
  • the optical fiber unit is centered on the tip part of one optical fiber connected to the LED pulse light source (wavelength 660 nm) in the light emitting part body.
  • Eight optical fibers connected to the PD in the main body of the detection light-receiving unit around the shaft are arranged with the tip portion coaxial (fiber unit FU-35FZ manufactured by Keyence Corporation).
  • a schematic cross-sectional view is shown in FIG. 9, where 101 is the tip of the light-emitting optical fiber (fiber strand diameter: 0.5 mm ⁇ 1), and 102 is the tip of the light-receiving fiber (fiber strand diameter: 0. 265 mm x 8), and a reflective lens F-2HA is attached to the tip of the fiber unit.
  • this light emitting / receiving unit is connected to an arithmetic processing unit (Keyence Corporation, digital fiber amplifier FS-N11MN) attached to the digital display unit to form one unit.
  • the optical fiber unit is arranged on the outside of the culture tank so as to be at a position 30 mm from the level of the bottom surface thereof, and at a position where the amount of received light and the like is maximum horizontally toward the reflecting mirror.
  • the turbidity measuring device used in the present embodiment is an integral type (coaxial type) of the light emitting unit and the light receiving unit for detection, and therefore does not require angle adjustment between the light emitting unit and the light receiving unit for detection.
  • it since it is structurally simple as an apparatus, it is convenient and suitable.
  • Example 4 (light receiving part: LED pulse light source, light emitting part / light receiving part integrated type) Except that the yeast cell concentration is as described in Table 4, the turbidity of the yeast suspension having a known yeast cell concentration is transmitted from the direction of the reflector caused by the irradiation light in the same manner as in Example 3. It was measured as a light receiving value of light such as light.
  • the turbidity measurement of the present invention can be applied as a transmitted light measurement method, a transmitted light / scattered light measurement method, or a scattered light measurement method depending on the turbidity of the culture solution. It was.
  • Table 4 for cell concentrations of 1.0 ⁇ 10 4 to 4.0 ⁇ 10 7 , the transmittance and absorbance are shown for reference.
  • the turbidity of the culture solution can be continuously measured over a wide range by measuring the turbidity as a light reception value of light from the direction of the reflector using the turbidity measuring apparatus of the present invention, It is extremely effective for time-dependent management of bacterial cell concentration and the like, such as grasping the growth process in the culture of cells.
  • Example 5 (light emitting part: LED pulse light source, light emitting part / light receiving part integrated type, bubble removing means)
  • the same turbidity measuring apparatus as in Example 3 was used, but the distance from the inner surface of the transparent part to the reflecting mirror was 15 mm (optical path length: 30 mm), and the following cylindrical shape was formed between the inner surface of the transparent part and the reflecting mirror: A device with and without a bubble removing means (enclosed near the optical path) was used.
  • Bubble removing means a cylinder made of a 30 mesh stainless steel net with a diameter of 20 mm and a length (height) of 15 mm. One end face is brought into contact with the inner surface of the transparent part, and the other end face is brought into contact with and fixed to a stainless steel reflecting mirror (a notch is formed as a bubble outlet on the upper part of the reflecting mirror).
  • the received light value from the direction of the reflector caused by the irradiated light is 100 to 130 times per minute. Moving and shaking vigorously, stable measurement was impossible.
  • the turbidity is measured using a turbidity measuring apparatus equipped with the bubble removing means, as apparent from Table 5 and FIG. 12, the culture solution is not greatly affected by bubbles. The turbidity could be measured accurately.

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Abstract

La présente invention concerne une technique de mesure optique de la turbidité d'une solution de culture dans une cuve de culture depuis l'extérieur de la cuve de culture. La présente invention concerne un dispositif de mesure de la turbidité conçu pour mesurer la turbidité d'une solution de culture depuis l'extérieur d'une cuve de culture, le dispositif de mesure de turbidité comprenant : un élément transparent placé dans une paroi de cuve d'une cuve de culture ; un miroir réfléchissant placé dans une solution de culture dans la cuve de culture ; un émetteur de lumière pour projeter de la lumière sur le miroir réfléchissant à travers l'élément transparent, l'émetteur de lumière étant placé à l'extérieur de la cuve de culture ; et un récepteur de lumière de détection pour recevoir la lumière venant de la direction du miroir réfléchissant à travers l'élément transparent.
PCT/JP2011/056955 2011-03-23 2011-03-23 Dispositif de mesure de turbidité WO2012127650A1 (fr)

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WO2020239823A1 (fr) * 2019-05-28 2020-12-03 Bifrost Biolabs Ivs Dispositif de mesure d'un paramètre dans un liquide
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WO2015033715A1 (fr) 2013-09-09 2015-03-12 株式会社日立製作所 Appareil de culture cellulaire et procédé associé
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WO2017141394A1 (fr) * 2016-02-18 2017-08-24 エイブル株式会社 Dispositif de réaction pour effectuer une centrifugation
JP2017150846A (ja) * 2016-02-22 2017-08-31 栗田工業株式会社 接液部材への付着物検出装置及び検出方法
WO2020239823A1 (fr) * 2019-05-28 2020-12-03 Bifrost Biolabs Ivs Dispositif de mesure d'un paramètre dans un liquide
WO2021019228A1 (fr) * 2019-07-29 2021-02-04 Imperial College Innovations Limited Procédé et appareil permettant de surveiller en temps réel la production d'un matériau dans une dispersion liquide
WO2021131555A1 (fr) * 2019-12-25 2021-07-01 味の素株式会社 Procédé pour la production d'un produit de fermentation et dispositif de détection à utiliser dans celui-ci

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