Incubation apparatus

An incubation apparatus, including a temperature-controlled room adjusted to be a predetermined environment condition, and incubating a sample of an incubation container inside the temperature-controlled room, includes a carrying apparatus, an imaging section, and an image analyzing section. The carrying apparatus carries the incubation container in the temperature-controlled room. The imaging section photographs a whole of the incubation container inside the temperature-controlled room. The image analyzing section analyzes an operation state of the incubation apparatus or an incubating environment state of the sample based on a total observing image of the incubation container photographed at the imaging section, and outputs an error signal notifying an abnormality of the operation state or the incubating environment state in accordance with the analysis result.

CROSS REFERENCE TO RELATED APPLICATION

This application is a U.S. National Stage application claiming the benefit of prior filed International Application Number PCT/JP2006/322458, filed Nov. 10, 2006, in which the International Application claims a priority date of Nov. 11, 2005 based on prior filed Japanese Application Number 2005-327469, the entire contents of which are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to an incubation apparatus incubating a sample of an incubation container in a temperature-controlled room which is adjusted to be a predetermined environment condition.

BACKGROUND ART

Conventionally, an incubation apparatus including a temperature-controlled room is generally used to incubate various kinds of microorganisms and cells. In general, for example, a sensor detecting environment conditions such as temperature, humidity, and carbon dioxide concentration, and an environment adjusting apparatus adjusting the above-stated respective parameters are disposed in the temperature-controlled room of the incubation apparatus, and therefore, inside the temperature-controlled room is adjusted to be a predetermined environment condition.

Besides, an incubation apparatus, distinguishing an incubation state of a sample by measuring an area and the number of the cells from a photographed image to perform processes such as an exchange of culture, is disclosed in Patent Document 1.

However, an object of the incubation apparatus in the above-stated Patent Document 1 is just to improve a workability involved in a subculture, and an analysis object thereof is only a state of the sample inside an incubation container. Consequently, there has been a problem in a point that it is impossible to detect an abnormal state caused by a change of an environment state surrounding the incubation container, a trouble of the incubation apparatus, and so on in the above-stated Patent Document 1. For example, a possibility in which the incubation state of the sample may deteriorate in accordance with passage of time when there is a scattering of culture from other incubation containers or a trouble of an incubation apparatus, and so on. Accordingly, a means capable of detecting such abnormal state in early time has been desired.

DISCLOSURE

Problems to be Solved

The present invention is made to solve the problems of the above-stated conventional art, and an object thereof is to provide an incubation apparatus capable of detecting abnormal states caused by a change of an environment state surrounding an incubation container, a trouble of the incubation apparatus, and so on.

Means for Solving the Problems

A first invention is an incubation apparatus including a temperature-controlled room adjusted to be a predetermined environment condition, and incubating a sample of an incubation container inside the temperature-controlled room, the incubation apparatus includes: a carrying apparatus, an imaging section, and an image analyzing section. The carrying apparatus carries the incubation container in the temperature-controlled room. The imaging section photographs a whole of the incubation container inside the temperature-controlled room. The image analyzing section analyzes one of an operation state of the incubation apparatus and an incubating environment state of the sample based on a total observing image of the incubation container photographed by the imaging section, and outputs an error signal notifying one of an abnormality of the operation state and the incubating environment state in accordance with the analysis result.

In a second invention according to the first invention, the image analyzing section detects an abnormal operation of the carrying apparatus based on a positional displacement of the incubation container extracted from the total observing image, and outputs the error signal when the abnormal operation is detected.

In a third invention according to the first or the second invention, the image analyzing section detects dew condensation inside the incubation container based on a change of a transmitted light amount inside the incubation container, and outputs the error signal when the dew condensation is detected.

In a fourth invention according to any one of the first to the third inventions, the image analyzing section estimates a culture amount inside the incubation container based on luminance and a ratio of respective color components inside the incubation container, and outputs the error signal when an estimated value of the culture amount deviates from a setting range.

In a fifth invention according to any one of the first to the fourth inventions, the image analyzing section detects a scattering of a culture by extracting at least one of the color component outside of the incubation container and a contour from the total observing image, and outputs the error signal when there is the scattering of the culture.

In a sixth invention according to any one of the first to the fifth inventions, the image analyzing section detects an occurrence of mold inside the incubation container based on the transmitted light amount and color information inside the incubation container, and outputs the error signal when the mold occurs.

In a seventh invention according to any one of the first to the sixth inventions, the carrying apparatus carries the incubation container corresponding to the total observing image out of the temperature-controlled room when the error signal is input.

In a eighth invention according to any one of the first to the sixth inventions, further includes: a notifying device performing a warning operation to outside of the incubation apparatus when the error signal is input.

In a ninth invention according to any one of the first to the eighth inventions, further includes: a microscope unit photographing a sample observing image in which a sample inside the incubation container is microscopic observed in every predetermined time when the error signal is not output; and a recording section recording the sample observing image.

Effect

According to an incubation apparatus of the present invention, abnormal states caused by a change of an environment state surrounding an incubation container, a trouble of the incubation apparatus, and so on can be detected.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Configuration of Incubator in Present Embodiment

Hereinafter, an incubator (incubation apparatus) in the present embodiment is described in detail with reference to the drawings.

An incubator11has a first casing12performing an incubation of a sample and a second casing13constituting a controller. The first casing12is used under a state disposed on the second casing13.

Here, configurations of incubation containers14performing an incubation of a sample are shown inFIG. 10(A) toFIG. 10(C). As the incubation container14in the present embodiment, a well plate, a flask, a dish, and so on are used. A liquid culture including a pH indicator such as phenol red and a sample (cells and so on) being an object of culture are accommodated in this incubation container14.

The above-stated incubation container14is handled while being mounted on a holder15in a transparent tray state. Support pieces16are respectively formed outward at both side surfaces of this holder15. Besides, an identification marker17performing an identification of the incubation container is put on the holder15. As an example of this identification marker17, for example, a two-dimensional code such as a QR code (a registered trademark), a bar code, and so on can be cited.

Next, a configuration of the first casing12is described based onFIG. 2toFIG. 4. A temperature-controlled room21covered with a heat insulator is formed inside the first casing12. This temperature-controlled room21is connected to the outside via a front opening22formed at a front side of the first casing12and a carrying in/out port23formed at a left side surface of the first casing. The front opening22of the first casing is openably/closably locked out by a front door24which is a single door or a double door. Incidentally, an example in which the front door24is constituted by the double door is shown inFIG. 2toFIG. 4. Besides, the carrying in/out port23is set to be approximately a size in which the incubation container14is capable of passing through. This carrying in/out port23is openably/closably locked out by an automatic door26sliding by a drive mechanism25. Incidentally, the automatic door26may be altered by a door opening/closing manually. Further, an opening27is formed at a bottom of the first casing12at a right-leaning position viewed from a front side. Incidentally, a later-described observation unit (38) is disposed inside the temperature-controlled room21via the opening27.

A temperature adjuster31, an atomizer32, a gas introducing section33, and an environment sensor unit34are accommodated in the temperature-controlled room21of the first casing12. The temperature adjuster31includes a Peltier element, and a heating or a cooling by a Peltier effect is performed by inverting an energizing polarity of the Peltier element. Incidentally, the temperature adjuster31may be altered by a well-known apparatus, for example, having a constitution such as a combination of a heater unit and a refrigerant circulation system.

Plural temperature adjusters31are accommodated at respective wall surfaces of the temperature-controlled room21, and each of the temperature adjusters31can perform a temperature control independently. The atomizer32adjusts humidity inside the temperature-controlled room21by spraying inside the temperature-controlled room21. The gas introducing section33is connected to a carbon dioxide cylinder (not-shown). This gas introducing section33introduces carbon dioxide into the temperature-controlled room21, to adjust carbon dioxide concentration inside the temperature-controlled room21. The environment sensor unit34detects the temperature, the humidity, and the carbon dioxide concentration inside the temperature-controlled room21.

Besides, a stocker35, a container carrying in/out mechanism36, a container carrying mechanism37, and an observation unit38are accommodated inside the temperature-controlled room21of the first casing12.

The stocker35is disposed at left side of the temperature-controlled room21viewed from the front side of the first casing12. The stocker35is divided vertically by plural shelves. The incubation container14can be accommodated horizontally in the stocker35. Besides, a lowermost shelf of the stocker35corresponds to a position of the carrying in/out port23of the first casing12. A space disposing the container carrying in/out mechanism36is formed at the lowermost shelf of the stocker35.

The container carrying in/out mechanism36is provided at the lowermost shelf of the stocker35. The container carrying in/out mechanism36has a carrier table36acapable of mounting a holder, and a motor unit36breciprocating the carrier table36atoward outside of the carrying in/out port23.

The container carrying mechanism37is disposed at a center of the temperature-controlled room21viewed from the front side of the first casing12. The container carrying mechanism37has a rectangular base table41, a vertical frame42extending in a longitudinal direction, and a carrier arm section43supporting the holder15.

The vertical frame42is attached to the base table41movably in a forward and backward direction (Y direction). A first motor44driving the vertical frame42in the Y direction is fixed at an outside of the base table41. Besides, a position in the Y direction of the vertical frame42is detected by a position sensor (not-shown) attached to the first motor44.

The vertical frame42is constituted by two guide rails disposed in parallel. The carrier arm section43is attached between the vertical frame42movably in a longitudinal direction (Z direction). This carrier arm section43moves by a rotation of a screw shaft (not-shown) housed in the vertical frame42. Further, a second motor45driving the carrier arm section43in the Z direction is attached to the vertical frame42at the base table41side. Incidentally, a position in the Z direction of the carrier arm section43is detected by a position sensor (not-shown) attached to the second motor45.

The carrier arm section43has a container support section46, a slide mechanism section47, and a third motor48. A main body of the container support section46is set to be a little wider than a whole width of the holder15including the support pieces16. At both side edges of the container support section46, a pair of hook claws46aare disposed to face at a lower side. Tip portions of the respective hook claws46aface toward inside of the container support section46, and a mutual interval between the tip portions of the hook claws46ais set to be slightly larger than the width of the main body portion of the holder15excluding the support pieces16. Accordingly, it is constituted such that the container support section46is able to support the holder15by an engagement between the support pieces16and the hook claws46a.

The slide mechanism section47is disposed at an upper surface side of the container support section46. This slide mechanism section47has a nut section (not-shown) screwing with the above-stated screw shaft. The slide mechanism section47slides the container support section46in a horizontal direction (X direction) by a drive of the third motor48. A transfer of the holder15mounting the incubation container14becomes possible between the stocker35, the container carrying in/out mechanism36, or the observation unit38and the container carrying mechanism37by an operation of the slide mechanism section47as stated above. Incidentally, a position in the X direction of the container support section46is detected by a position sensor (not-shown) attached to the third motor48.

The observation unit38is disposed at right side of the temperature-controlled room21viewed from the front side of the first casing12. This observation unit38is disposed while fit into the opening27at the bottom of the first casing12. This observation unit38has a stage39, an arm38athrowing out toward upward of the stage39, and a main body portion38b. The stage39and the arm38aare disposed inside the temperature-controlled room21of the first casing12, on the other hand, the main body portion38bis accommodated in the second casing.

FIG. 9is a schematic diagram showing a configuration of the observation unit38. The observation unit38has the stage39, a first illumination section51and a second illumination section52, a microscopic observation system53having an LED unit53a, a container observation system54, and an image processing section55.

The stage39is constituted by a translucent material, and the incubation container14is mounted thereon together with the holder15. Besides, the first illumination section51and the LED unit53aare disposed at the arm38a, to illuminate the incubation container14from an upper side of the stage39. On the other hand, the second illumination section52is housed in the main body portion38b, to illuminate the incubation container14from a lower side of the stage39.

The microscopic observation system53is housed in the main body portion38b, and it has a microscopic optical system and an image pickup device (both are not shown). This microscopic observation system53photographs an image in which a sample is microscopically observed with transmitted light of the incubation container14by the illumination of the LED unit53a(a sample observing image).

The container observation system54is accommodated in the arm38a, and has an imaging optical system and an image pickup device (both are not shown). This container observation system54photographs a total observing image of the incubation container14by illumination light of the first illumination section51or the second illumination section52.

The image processing section55generates a color image data of the sample observing image or the total observing image by performing an A/D conversion of outputs of the microscopic observation system53and the container observation system54, and performing various image processing. Besides, the image processing section55performs an image analysis on the image data of the total observing image.

Next, a configuration of the second casing13is described. The above-stated main body portion38bof the observation unit38and a control unit are stored in the second casing13.

The control unit has a CPU61, an operation instruction section62, a database section63, a display panel64, and a communication section65.

The CPU61is connected to the drive mechanism25, the temperature adjuster31, the atomizer32, the gas introducing section33, the environment sensor unit34, the container carrying in/out mechanism36, the container carrying mechanism37, the observation unit38, the operation instruction section62, the database section63, the display panel64, and the communication section65. The CPU61controls the above-stated respective sections in accordance with a predetermined sequence program. Incidentally, the CPU61has a clock function for a schedule management.

The operation instruction section62has an input device such as a keyboard, and operates each section of the incubator11via the CPU61. Namely, the CPU61performs an adjustment of an environment condition inside the temperature-controlled room21, a carrying in/out of the incubation container14into/out of the temperature-controlled room21, an observation of the sample of the incubation container14, a carry of the incubation container14inside the temperature-controlled room21, and so on based on an input from the operation instruction section62. Here, both of an instruction by a direct input of a user and an instruction set by a program in advance are included in the instructions of the operation instruction section62.

Management data of the incubation container14, various data required for an image analyzing, and so on are recorded on the database section63. For example, identification codes of the incubation containers14, types and shapes of the incubation containers14, accommodated positions of the incubation containers14, data of an observation schedule of each incubation container14set at the operation instruction section62, and so on are included in the above-stated management data. Besides, the data of the accommodated positions of the incubation containers14may be registered by an input of the user. Otherwise, the CPU61may automatically set the accommodated positions of the incubation containers14by reading the identification markers17of the holders15with a camera and so on.

Besides, it is possible to record the image data of the sample observing image and the total observing image in the database section63. Here, the image data recorded on the database section63are recorded while respectively being corresponded to index data containing identification information of the incubation containers14and photographed dates and times. Further, a change history of the environment condition (temperature, humidity, carbon dioxide concentration) inside the temperature-controlled room21can also be recorded on the database section63.

Values of the environment condition of the temperature-controlled room21output from the CPU61, and a warning at the time of occurrence of abnormality are displayed on the display panel64. The communication section65controls a data transmission/reception between an external computer (not-shown) and so on, complying with a well-known wireless or wired communication protocol.

(Operations of Incubator in Present Embodiment)

Hereinafter, operations of the incubator specific to the present embodiment are described following a flowchart inFIG. 11. The incubator11of the present embodiment automatically performs the photographing of the sample observing image and the total observing image based on the observation schedule which is set in advance. The incubator11then detects the abnormal states resulting from a change of the environment condition surrounding the incubation container14, a trouble of the incubation apparatus, and so on based on the total observing image. Incidentally, the actual incubator11manages the plural incubation containers14simultaneously based on the observation schedule by each incubation container, though the description is made while limiting to a management of one incubation container as a matter of convenience in an example inFIG. 11.

Step101: The CPU61determines whether an observation time of the incubation container14has come or not by comparing the observation schedule in the database section63and the current date and time. When it is the observation time (YES side), the process goes to S102. On the other hand, when it is not the observation time of the incubation container14(NO side), the CPU61stands ready for the observation time of the incubation container.

Step102: The CPU61acquires the data of the accommodated position of the incubation container14which is an observation object from the database section63. Next, the CPU61instructs the container carrying mechanism37to carry the incubation container14which is the observation object from the stocker35to the observation unit38. The container carrying mechanism37carries out the indicated incubation container14from the stocker35, and mounts the incubation container14on the stage39of the observation unit38.

Step103: The CPU61determines whether the photographing of the total observing image is performed or not in the current photographing based on the observation schedule. When the total observing image is photographed (YES side), the process goes to S104. On the other hand, when only the sample observing image is photographed (NO side), the process goes to S111.

Step104: The CPU61instructs the observation unit38to photograph the total observing image. The observation unit38illuminates the incubation container14by lighting at least one of the first illumination section51and the second illumination section52, and photographs the total observing image of the incubation container14with the image pickup device of the container observation system54.

Step105: The image processing section55of the observation unit38performs the image analysis for the total observing image. The image processing section55outputs an error signal when it is in the abnormal state from a result of the above-stated image analysis, and outputs a normal signal when it does not correspond to the abnormal state. Here, in S105, it is possible to perform the following: (1) Estimation of a culture amount; (2) Detection of a scattering of a culture; (3) Detection of mold inside the incubation container; (4) Detection of a positional displacement of the incubation container; and (5) Detection of dew condensation inside the incubation container. Incidentally, concrete contents of the above-stated (1) to (5) are respectively described later.

Step106: The CPU61determines whether the error signal is input from the image processing section55of the observation unit38or not. When the error signal is input (YES side), the process goes to S107. On the other hand, when the normal signal is input (NO side), the process goes to S110.

Step107: The CPU61determines whether a setting in which the incubation container14is discharged from the temperature-controlled room21is made or not when the error signal is input. When the incubation container14is discharged (YES side), the process goes to S108. On the other hand, when the incubation container14is not discharged (NO side), the process goes to S109.

Here, the setting whether the incubation container14is discharged or not can be set individually by a user in advance, by each of the cases of (1) to (5) in S105. For example, when the scattering of the culture or the mold are detected, it is especially desirable to discharge the incubation container14which is the observation object from the temperature-controlled room rapidly, because there is an adverse affect on the sample of the other incubation containers14. On the other hand, when the estimated culture amount is not appropriate or when something is wrong with the container carrying mechanism37, the temperature adjustor31, or the like, there is a possibility that it is not an appropriate treatment to discharge the incubation container14out of the temperature-controlled room.

Step108: The CPU61instructs the container carrying in/out mechanism36and the container carrying mechanism37to discharge the incubation container14which is the observation object out of the temperature-controlled room. The container carrying mechanism37transfers the indicated incubation container14from the stage39to the container carrying in/out mechanism36. The CPU61opens the automatic door26of the carrying in/out port23, and the container carrying in/out mechanism36discharges the incubation container14out of the temperature-controlled room21.

Step109: The CPU61performs warning operations notifying the occurrence of the abnormality. Here, as the warning operations, for example, a warning display on the display panel64, a transmission of an error notification mail to the external computer by the communication section65, an audio output by a not-shown buzzer, and so on can be cited. Incidentally, contents of these warning operations can be set by the user in advance by specifying in the operation instruction section62.

Step110: The CPU61determines whether the photographing of the sample observing image is performed or not in the current photographing based on the observation schedule. When the sample observing image is photographed (YES side), the process goes to S111. On the other hand, when the sample observing image is not photographed (NO side), the process goes to S112.

Step111: The CPU61instructs the observation unit38to photograph the sample observing image. The observation unit38illuminates the incubation container14with the LED unit53a, and photographs the sample observing image with the image pickup device of the microscopic observation system53. Incidentally, the image data of the sample observing image is recorded on the database section63while being corresponded to the above-stated index data.

Step112: The CPU61instructs the container carrying mechanism37to carry the incubation container14which is the observation object from the observation unit38to the stocker35. The container carrying mechanism37carries out the indicated incubation container14from the stage39, and returns the incubation container14to a predetermined position of the stocker35. After that, the CPU61returns to S101to repeat the above-stated operations.

(Estimation of Culture Amount)

Next, the “estimation of the culture amount” in the above-stated S105is described in detail following a flowchart inFIG. 12. The reason why the “estimation of the culture amount” is performed at S105is because judgments of a culture exchange or an incubation stop become easier if there is information whether the culture amount is within an appropriate range or not.

Step201: The image processing section55performs an extraction process of edges for the color image data of the total observing image (S104), to extract a contour of the incubation container14.

Step202: The image processing section55performs a pattern matching between the contour data of the incubation container14(S201) and a contour template of the incubation container14, to specify an area in which an inside of the incubation container14is photographed (an incubation area of the sample). When one incubation container has plural individual small containers, the image processing section55respectively specifies the incubation areas corresponding to the respective small containers. Incidentally, in this case, the description is made on the assumption that similar operations are performed by each of the individual incubation areas by the following steps.

Step203: The image processing section55determines plural measurement points from among the incubation areas of the sample (S202). Positions of the measurement points in the incubation areas may be determined in advance. The image processing section55acquires signal values of three-color components of red, green, blue respectively by each of the measurement points based on the color image data of the total observing image. Incidentally, the signal values of the respective color components become larger as luminance of the color component at the measurement point becomes higher.

Step204: The image processing section55asks a ratio (B/G ratio) between the signal value of the blue component and the signal value of the green component by each measurement point. The image processing section55estimates a pH value at each measurement point based on the B/G ratio by, for example, the following expression (1).
B/G=0.56×pH^2−7.48×pH+25.4  (1)

The above-stated expression (1) is a quadratic function approximately showing a relation of a change between the pH value and the B/G ratio. The “B/G” in the above expression shows the B/G ratio. Besides, the “pH” in the above expression shows the estimated pH value.

Here, the pH value is estimated based on the B/G ratio in the present embodiment because of the following reasons. The present inventors obtained the following knowledge as a result of researches as for the relation between a pH indicator (phenol red) added to the culture and the signal value of each color component of the image.

When a change of color of the phenol red and the ratio of the signal values of the respective color components are compared on the assumption that concentrations of the cultures are the same, the ratio of the blue component increases in accordance with an increase of the pH value, on the other hand, the ratio of the green component decreases. Besides, if the pH value changes, there is not a very large change in the signal value of the red component. Namely, it turns out that a determination of the pH value by the color of the phenol red is synonymous with a determination of the ratio between blue and green. Besides, the color of the phenol red (the ratio between blue and green) non-ambiguously changes in accordance with the pH value. The image processing section55estimates the pH value inside the incubation container14by the B/G ratio based on the above-stated theory.

Step205: The image processing section55estimates the concentration of the culture at the measurement point based on the signal value of the red component (S203) and the estimated pH value (S204). Concretely speaking, data showing a correspondence between “the signal value of the red component” and “the concentration of the culture”, and correction data by the pH value are stored in the database section. The image processing section55reads the above-stated respective data from the database section63, and calculates the concentration of the culture from the signal value of the red component based on the above-stated data.

Here, the concentration of the culture is estimated based on the signal value of the red component because of the following reasons. When the images of the incubation containers14having a difference in the concentrations of the cultures are compared on the assumption that the pH values are the same, the color of the culture becomes dark as the concentration of the culture becomes higher. Accordingly, the signal value of the red component becomes large in proportion to the increase of the concentration of the culture. Consequently, it becomes possible to estimate the concentration of the culture by the signal value of the red component. At this time, a change rate between the signal value of the red component and the concentration of the culture is different in accordance with the pH values, and therefore, the image processing section55performs the correction by the pH value.

Step206: The image processing section55estimates the culture amount inside the incubation container14based on the container shape specified from the type of the incubation container14, and the concentration of the culture (S205). Incidentally, the concentration of the culture in this S206is determined by the image processing section55based on measurement results at the plural measurement points.

Step207: The image processing section55determines whether the estimated value of the culture amount inside the incubation container14(S206) is within a predetermined range or not. When the estimated value is within the predetermined range (YES side), the process goes to S208. On the other hand, when the estimated value deviates from the predetermined range (NO side), the process goes to S209.

Step208: In this case, the image processing section55outputs a normal signal showing that “the culture amount is appropriate” to the CPU61.

Step209: In this case, the image processing section55outputs an error signal showing that “the culture amount is little” or “the culture amount is much” to the CPU61.

(Detection of Scattering of Culture)

Next, the “detection of the scattering of the culture” in the above-stated S105is described in detail following a flowchart inFIG. 13. A possibility in which a contamination to the other incubation containers may occur is high when there is the scattering of the culture. Accordingly, it is necessary to detect the scattering of the culture in early time. Incidentally, S301inFIG. 13corresponds to the above-stated S201, and therefore, a redundant description is not given.

Step302: The image processing section55performs the pattern matching between the contour data of the incubation container14(S301) and the contour template of the incubation container14, and specifies the area in which the inside of the incubation container is photographed (the incubation area of the sample). Incidentally, when one incubation container14has the plural individual small containers, the above-stated incubation area is specified by each small container. The image processing section55sets a portion excluding the incubation area of the sample inside of the incubation container14as a determination area (refer toFIG. 14).

Step303: The image processing section55detects an area showing the color component of the pH indicator in the determination area. Incidentally, when there is the area having the above-stated color component within the determination area, it can be judged that there is the scattering of the culture.

Step304: The image processing section55determines whether the scattering of the culture is detected from the determination area or not. When it is detected (YES side), the process goes to S305. On the other hand, when it is not detected (NO side), the process goes to S306.

Step305: In this case, the image processing section55outputs an error signal showing that “there is the scattering of the culture” to the CPU61.

Step306: On the other hand, in this case, the image processing section55outputs a normal signal showing that “the scattering of the culture is not recognized” to the CPU61.

(Detection of Mold Inside Incubation Container)

Next, the “detection of the mold inside the incubation container” in the above-stated S105is described in detail following a flowchart inFIG. 15. When the mold is occurred inside an incubation container, a possibility in which the mold increases also in the other incubation containers is high. Accordingly, it is necessary to detect the occurrence of the mold in early time.

Here, when the detection of the mold inside the incubation container14is performed, it is necessary to photograph the total observing image by illuminating the incubation container14with the second illumination section52from an opposite side of the container observation system54in S104. Incidentally, S401and S402inFIG. 15are respectively corresponding to the above-stated S201and S202, and therefore, redundant descriptions are not given.

Step403: The image processing section55calculates a transmitted light amount in all over the incubation area of the sample (S402). This data of the transmitted light amount in S403becomes a reference when the image processing section55determines a decrease of the transmitted light amount caused by the mold.

Step404: The image processing section55scans the incubation area of the sample (S402) in a constant direction, to detect an area where the transmitted light amount decreases more than a threshold value. The image processing section55confirms color information of the detected area, and specifies the occurrence area of the mold. When the mold occurs, the transmitted light amount of the culture decreases significantly, and a color change such that the culture becomes white can be recognized if the mold is, for example, mildew. Accordingly, the image processing section55can detect the occurrence of the mold by the above-stated operations.

Step405: The image processing section55determines whether the mold is detected inside the incubation container14or not. When the mold is detected (YES side), the process goes to S406. On the other hand, when the mold is not detected (NO side), the process goes to S408.

Step406: The image processing section55calculates an area ratio of the mold relative to the incubation container14from a size of the occurrence area of the mold (S404) and the shape of the incubation container14. Incidentally, there is a case when this process in S406is omitted by the image processing section in accordance with a setting at the operation instruction section62.

Step407: The image processing section55outputs an error signal showing that “the mold is occurred inside the incubation container” to the CPU61. At this time, the data of the area ratio of the mold in S406may be simultaneously output to the CPU61.

Step408: On the other hand, in this case, the image processing section55outputs a normal signal showing that “the occurrence of the mold is not recognized inside the incubation container” to the CPU61.

(Detection of Positional Displacement of Incubation Container)

Next, the “detection of the positional displacement of the incubation container” in the above-stated S105is described in detail following a flowchart inFIG. 16. A possibility becomes high, in which an accident may occur at a carrying time of the incubation container under a state in which the container carrying mechanism does not operate normally. Accordingly, it becomes necessary to confirm the operation state of the container carrying mechanism by a presence/absence of the positional displacement of the incubation container. Incidentally, S501inFIG. 16corresponds to the above-stated S201, and therefore, a redundant description is not given.

Step502: The image processing section55reads data of a positional displacement determination template from the database section63. This positional displacement determination template corresponds to the contours of the incubation containers14at normal container setting positions. Consequently, the contours of the incubation containers14extracted from the total observing image and the contours of the positional displacement determination template are matched under a state in which the container carrying mechanism37operates normally. The image processing section55performs a matching between the contour data of the incubation container14(S501) and the positional displacement determination template, and acquires a displacement amount between the normal container setting positions and the current positions of the incubation containers (refer toFIG. 17).

Step503: The image processing section55determines whether the above-stated displacement amount (S502) falls within a range of a tolerance or not. When the displacement amount falls within the range of the tolerance (YES side), the process goes to S504. On the other hand, when the displacement amount does not fall within the range of the tolerance (NO side), the process goes to S505.

Step504: In this case, the image processing section55outputs a normal signal showing that “the container carrying mechanism operates normally” to the CPU61.

Step505: On the other hand, in this case, the image processing section55outputs an error signal showing that “the operation of the container carrying mechanism is abnormal” to the CPU61.

(Detection of Dew Condensation Inside Incubation Container)

Next, the “detection of the dew condensation inside the incubation container” in the above-stated S105is described in detail following a flowchart inFIG. 18. The dew condensation occurs inside the incubation container when there is temperature unevenness resulting from a trouble of a temperature adjustor and so on, and an operation of a motor and so on. If the dew condensation is detected, it becomes possible to confirm the incubation state of the sample, and to improve an incubation accuracy further more.

Here, when the detection of the dew condensation inside the incubation container is performed, it is necessary to photograph the total observing image by illuminating the incubation container14with the second illumination section52from the opposite side of the container observation system54in S104. Besides, in the detection of the dew condensation inside the incubation container14, an analysis is performed by using plural total observing images photographed at different times. Incidentally, S601and S602inFIG. 18respectively correspond to the above-stated S201and S202, and therefore, redundant descriptions are not given.

Step603: The image processing section55calculates the transmitted light amount in all over the incubation area of the sample (S602). Data of the transmitted light amount in this S603becomes a reference when the image processing section55determines a decrease of the transmitted light amount caused by the dew condensation. Besides, the image processing section55acquires a luminance level of a measurement reference point (not-shown) existing at a predetermined position of the holder in the total observing image.

Step604: The image processing section55scans the incubation area of the sample (S602) in the constant direction, and detects an area where the transmitted light amount decreases more than a threshold value. Besides, the image processing section55records a detected result of this time and the luminance level of the measurement reference point to the database section63after the detection process in S604.

Step605: The image processing section55reads the detected result of a previous time from the database section63, and performs a comparison with the detected result of this time (S604). At this time, the image processing section55performs the comparison after normalizing the value of the transmitted light amounts between the plural images based on the luminance level of the measurement reference point.

Step606: The image processing section55determines whether the dew condensation occurs inside the incubation container14or not. Concretely speaking, the image processing section55determines that the dew condensation occurs when the decrease of the transmitted light amount becomes larger than the previous time. It goes without saying that the image processing section55may perform the determination by using the determination result before the previous time. Incidentally, a damping of the transmitted light amount in case of the dew condensation is smaller than the case of the mold, and the color change such as the mold does not occur, and therefore, it is fully possible to distinguish between both.

When the dew condensation occurs (YES side), the process goes to S607. On the other hand, when the dew condensation does not occur (NO side), the process goes to S608.

Step607: In this case, the image processing section55outputs an error signal showing that “the dew condensation occurs” to the CPU61. At this time, the image processing section55may estimate a droplet amount inside the incubation container by the change of the luminance value, the estimation of the culture amount, and so on in the total observing image.

Step608: On the other hand, in this case, the image processing section55outputs a normal signal showing that “the dew condensation does not occur” to the CPU61.

(Effect of Present Embodiment)

The incubator11of the present embodiment automatically detects the change of the culture amount, the occurrence of the mold, the scattering of the culture, and so on, and performs the carrying out of the incubation container14and the predetermined warning operations when something is wrong with the environment state surrounding the incubation container14. Accordingly, a manual labor of the confirmation of the incubation state of the sample is reduced significantly, and it becomes easy to keep the incubation state of the sample in the temperature-controlled room21in good state.

Besides, it is possible for the incubator11of the present embodiment to automatically detect the positional displacement of the incubation container14resulting from the abnormality of the container carrying mechanism37and the dew condensation of the incubation container14caused by the temperature unevenness of the temperature-controlled room21, and to perform the predetermined warning operations. Accordingly, it is possible to drastically suppress the occurrence of accidents resulting from the abnormality of the incubation apparatus.

Further, the incubator11of the present embodiment photographs the microscopic observed image of the sample in accordance with the predetermined schedule, and records the image data as a database. Accordingly, it becomes possible to significantly reduce the manual labor of the confirmation of the incubation condition of the sample, and to judge timings and so on of the culture exchange or the subculture by the user based on the image data.

(Supplemental Matters of Embodiment)

(1) Respective configurations of the incubator of the present invention are not limited to the above-stated embodiment. For example, the present invention can be applied to a multi-gas incubator capable of adjusting at least one of oxygen concentration or nitrogen concentration in addition to the carbon dioxide concentration. Besides, the adjustment of the humidity may be performed by a humidification water reservoir storing humidification water and a temperature adjustor controlling a water temperature of the humidification water reservoir (both are not shown).

(2) A macro observation system capable of microscopic observation with lower magnification than the microscopic observation system may be provided as the observation unit of the above-stated embodiment. When the microscopic observation is performed, two kinds of sample observing images by the macro observation system and the microscopic observation system may be photographed in a series of sequence. Besides, in the above-stated embodiment, the image analysis may be performed not by the observation unit but by the CPU.

(3) The incubator of the present invention may have a configuration selectively performing either of the estimation of the culture amount, the detection of the scattering of the culture, the detection of the mold, the detection of the positional displacement of the incubation container, and the detection of the dew condensation.

Besides, in the detection of the positional displacement of the incubation container and the detection of the scattering of the culture, it is possible to determine by using the extracted contours, and therefore, the total observing image may be a gray scale image when only these determination are performed. Incidentally, in the detection of the positional displacement of the incubation container, markers may be disposed at plural points of the incubation container to detect the abnormality of the container carrying mechanism from positional displacements of the markers.

(4) In S108of the above-stated embodiment, the discharge of the incubation container and the warning operation (S109) may be performed simultaneously. Besides, in S109of the above-stated embodiment, the incubation container may be temporary returned to the stacker after the warning operation.