JP2009002842A - Apparatus, method, and culture vessel for spectral inspection - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To perform spectral analysis of an sample without being affected by a culture medium, without sucking out the culture medium from a culture vessel. <P>SOLUTION: A device for spectral inspection comprises a stage 5 for mounting the culture vessel 4 which accommodates the sample 3 with the culture medium; a light source for generating near infrared light L which irradiate the sample 3 mounted on the stage 5; a sensor 7 for receiving the near infrared light L which is received by the sensor 7, and a section 8 for measuring spectroscopy by dispersing the near infrared light L received by the sensor 7, in which the stage 5 has the culture vessel 4 being inclined from the horizontal direction mountably. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a spectroscopic inspection apparatus, a spectroscopic inspection method, and a culture container for spectroscopic inspection.

  2. Description of the Related Art Conventionally, a spectroscopic analyzer is known that irradiates a culture solution containing cells to be measured with near-infrared light, and receives and analyzes reflected light or transmitted light from the culture solution (for example, Patent Document 1). reference.).

JP 2004-149144 A

  However, in the spectroscopic analysis apparatus of Patent Document 1, in order to irradiate the cells existing in the culture solution with light, the culture solution (mainly water) filled around the cells strongly absorbs light. There is. As a result, the signal from the cell itself is very small compared to the signal from the culture medium, which makes it difficult to quantify.

  In order to avoid this inconvenience, it is conceivable that the culture solution is sucked out from the culture vessel with a pipette to expose the cells prior to the examination. There is an inconvenience that the possibility of the nation increases and the workability is poor because the sucking work is required.

  The present invention has been made in view of the above-described problems, and is a spectroscopic inspection apparatus capable of performing spectroscopic analysis of a sample without being influenced by the culture solution without performing an operation of sucking out the culture solution from the inside of the culture vessel. An object of the present invention is to provide a spectroscopic inspection method and a culture container for spectroscopic inspection.

In order to achieve the above object, the present invention provides the following means.
The present invention includes a stage for placing a culture container that contains a sample together with a culture solution, a light source that generates near-infrared light for irradiating the sample in the culture container placed on the stage, and the light source A sensor that receives near-infrared light that is irradiated and reflected or transmitted from the sample, and a spectroscopic measurement unit that measures near-infrared light received by the sensor to measure wavelength characteristics, and the stage includes: Provided is a spectroscopic inspection apparatus configured such that the culture container can be placed while being inclined with respect to a horizontal direction.

  According to the present invention, when the culture vessel containing the culture medium and the sample is placed on the stage, the stage is configured so that the culture vessel can be placed with the culture vessel tilted with respect to the horizontal direction. When the culture vessel is tilted, the culture solution stored in the container moves downward along the bottom surface, and the sample growing while adhering to the bottom surface is exposed from the culture solution. In this state, when the near-infrared light is emitted from the light source, the emitted near-infrared light is irradiated onto the sample without passing through the culture solution, and the near-infrared light reflected or transmitted by the sample is received by the sensor, The wavelength characteristic is measured by spectroscopic measurement in the spectroscopic measurement unit. That is, the wavelength characteristic can be acquired without being affected by the culture solution stored in the culture vessel. Therefore, the signal from the sample can be easily quantified without performing the work of sucking out the culture solution from the culture vessel.

In the said invention, the said light source may be arrange | positioned so that near infrared light may be irradiated to the thickness direction of the sample in the said culture container.
By doing so, it is possible to accurately perform spectroscopic analysis of a sample whose thickness increases with growth.

Moreover, in the said invention, the said light source, a sensor, and a stage may be provided so that rocking | fluctuation integrally is possible around the substantially horizontal axis line.
In this way, the stage can be set in a horizontal state, the culture vessel can be placed, and the sample can be exposed from the culture solution in the culture vessel by swinging the stage around a substantially horizontal axis. it can. In this case, since the light source and the sensor are swung integrally with the stage, near infrared light is irradiated to the sample exposed from the culture solution by tilting the stage, and the reflected or transmitted near infrared light is detected by the sensor. Can be received.

  The present invention also includes an exposure step of exposing the sample from the culture solution by tilting a culture vessel that contains the sample together with the culture solution, and an irradiation step of irradiating the sample exposed from the culture solution with near infrared light. Provided is a spectroscopic inspection method comprising a light receiving step for receiving near-infrared light reflected or transmitted from a sample and a measurement step for measuring wavelength characteristics by splitting the received near-infrared light.

  According to the present invention, since the sample exposed from the culture medium is tilted and irradiated with near-infrared light to receive and measure the wavelength characteristics, the spectral analysis of the sample is not affected by the culture liquid. Can be performed with high accuracy.

In the said invention, the said irradiation step is good also as irradiating a near-infrared light along the optical axis along the thickness direction of the sample inclined with respect to the perpendicular direction by inclining a culture container.
By doing so, it is possible to accurately perform spectroscopic analysis of a sample whose thickness increases with growth.

  Further, the present invention provides a spectroscopic examination culture container having a recess capable of accommodating a sample together with a culture solution, wherein the recess has a flat bottom surface on which the sample can be adhered and cultured, Provided is a spectroscopic culture container having a shape having different areas across a straight line passing through the position of the center of gravity.

  According to the present invention, the culture medium and the sample are accommodated in the recess, and the bottom surface is set to be approximately horizontal, and the sample grows by being adhered to the flat bottom surface in the recess. When performing spectroscopic inspection, by inclining the bottom surface, the culture solution in the recess can be moved downward to expose the sample adhered to the bottom surface. In this case, since the bottom surface of the recess has a shape having different areas across the straight line passing through the center of gravity, the liquid surface of the culture solution is further lowered by inclining so that the larger area is on the lower side. The sample can be easily exposed by moving to the side. Therefore, it is possible to perform spectroscopic analysis of the sample without being affected by the culture solution.

  The present invention also relates to a spectroscopic culture container having a recess capable of accommodating a sample together with a culture solution, wherein the recess has a flat culture surface on which a sample can be adhered and cultured, and the culture An observation portion having a flat observation surface adjacent to the culture surface and disposed on the opposite side of the culture portion across the observation portion, the observation surface being substantially horizontal A culture container for spectroscopic examination is provided, comprising: a culture solution storage unit having a volume for storing the culture solution stored in the culture unit when arranged.

  According to the present invention, a sample and a culture solution are accommodated and cultured in a culture part provided in the recess, so that the sample is grown on a flat culture surface arranged substantially horizontally in the culture part. be able to. When performing spectroscopic inspection of the sample, the culture surface is inclined so that the culture portion is arranged above the observation portion, and the observation surface adjacent to the culture surface is made substantially horizontal, so that the culture portion As a result, the sample stored on the culture surface is allowed to flow out from the culture unit, and the sample adhered to the culture surface can be gradually moved to the observation surface. At this time, since the culture solution storage unit is adjacent to the lower side of the observation unit, the culture solution flowing out from the culture unit passes through the observation unit and flows into the culture solution storage unit. Thereby, since the sample moved to the observation part is exposed from the culture solution, the spectroscopic inspection can be performed without being influenced by the culture solution.

  The present invention also relates to a spectroscopic culture container having a recess capable of accommodating a sample together with a culture solution, wherein the recess has a flat culture surface on which a sample can be adhered and cultured, and the culture An observation part having a flat observation surface adjacent to the culture surface across the inclined surface that is inclined with respect to the culture surface of the part, and disposed on the opposite side of the culture unit across the observation part And a culture medium storage part having a volume for storing the culture medium stored in the culture part when the inclined surface is arranged substantially horizontally, the observation surface of the observation part, and the culture medium Provided is a spectroscopic examination culture vessel provided with a partition wall that prohibits the passage of a sample and allows the passage of a culture solution between the container and the container.

  According to the present invention, a sample and a culture solution are accommodated and cultured in a culture part provided in the recess, so that the sample is grown on a flat culture surface arranged substantially horizontally in the culture part. be able to. Then, when performing spectroscopic inspection of the sample, the culture surface is inclined so that the culture part is arranged above the observation part, and the sample adhered to the culture surface is passed through the inclined surface adjacent to the culture surface. And gradually move to the observation surface. Since a partition wall is provided between the observation surface of the observation unit and the culture solution storage unit, the sample is blocked by the partition wall, and the culture solution storage unit is arranged below the observation unit. Into the house. In this state, the sample is exposed to and placed on the observation surface from the culture medium by returning the tilt so that the observation surface is substantially horizontal. Can be done.

  According to the present invention, there is an effect that it is possible to perform spectroscopic analysis of a sample without being influenced by the culture solution without performing an operation of sucking out the culture solution from the inside of the culture vessel.

A spectroscopic inspection apparatus 1 according to an embodiment of the present invention will be described below with reference to FIG.
As shown in FIG. 1, the spectroscopic inspection apparatus 1 according to this embodiment includes a stage 5 on which a culture vessel 4 that houses a sample 3 together with a culture solution 2 is placed, and a culture vessel 4 that is placed on the stage 5. A light source 6 that irradiates the near-infrared light L to the sample 3 in the inside, a sensor 7 that detects the near-infrared light L irradiated from the light source 6 and transmitted through the sample 3, and the sensor 7 detects And a spectroscopic measurement unit 8 that obtains wavelength characteristics by splitting the near-infrared light L.

  The culture container 4 is, for example, a petri dish-like container having a substantially circular bottom surface 4a, and is made of a material such as transparent quartz or plastic that can transmit near-infrared light L. The sample 3 to be cultured is, for example, cartilage tissue that grows by adhering to the bottom surface 4a.

  The stage 5 is arranged to be inclined in one direction with respect to the horizontal direction. A through hole 5a that penetrates in the thickness direction is provided at a substantially central position of the stage 5. Furthermore, a stopper 5c that abuts the culture vessel 4 to be placed is provided below the placement surface 5b of the stage 5.

  The light source 6 is disposed so as to emit near-infrared light L from below the stage 5 through the through hole 5a of the stage 5 obliquely upward. The sensor 7 is disposed at a position facing the light source 6 with the stage 5 interposed therebetween.

  When the culture vessel 4 is placed on the placement surface 5 b of the stage 5 and the lower part of the culture vessel 4 is abutted against the stopper 5 c, the through-hole 5 a is closed by the culture vessel 4. The infrared light L passes through the through-hole 5a of the stage 5 and is irradiated to the culture vessel 4 so that it passes through the culture vessel 4 and the sample 3 accommodated in the culture vessel 4 and is received by the sensor 7. It has become.

A spectroscopic inspection method using the spectroscopic inspection apparatus 1 according to this embodiment configured as described above will be described below.
In order to perform spectroscopic analysis of the sample 3 using the spectroscopic inspection apparatus 1 according to the present embodiment, the sample 3 is accommodated in the culture vessel 4 together with the culture solution 2 and cultured under predetermined culture conditions in an incubator (not shown). The sample 3 is grown by performing the above, and the culture container 4 is taken out from the incubator and placed on the placement surface 5 b of the stage 5 at a predetermined point in the growth process.

  Although the placement surface 5b of the stage 5 is disposed to be inclined, since the stopper 5c is provided on the placement surface 5b, the lower end of the culture vessel 4 abuts against the stopper 5c and is fixed. As a result, when the culture vessel 4 containing the culture solution 2 and the sample 3 is placed on the placement surface 5b of the stage 5, the sample 3 in the culture vessel 4 is stored in the culture vessel 4 as shown in FIG. While maintaining the adhesive state on the bottom surface 4a, the culture solution 2 flows downward in the culture vessel 4 and accumulates in the lower part. As a result, the sample 3 in the culture vessel 4 is exposed from the culture solution 2.

  In this state, by operating the light source 6, the near-infrared light L from the light source 6 is emitted obliquely upward through the through hole 5 a of the stage 5, passes through the culture vessel 4 and passes through the sample in the culture vessel 4. 3 is irradiated. Since the sample 3 is exposed from the culture solution 2, the near-infrared light L passes through the sample 3 and is received by the sensor 7 without passing through the culture solution 2. The signal from the sensor 7 is sent to the spectroscopic measurement unit 8 to measure the wavelength characteristics, whereby the components contained in the sample 3 can be analyzed quantitatively.

  As described above, according to the spectroscopic inspection apparatus 1 according to the present embodiment, the sample 3 is removed from the culture medium 4 simply by placing the culture vessel 2 containing the culture solution 2 and the sample 3 on the placement surface 5 b of the stage 5. The sample 3 can be subjected to spectroscopic analysis without being influenced by the culture solution 2. Therefore, there is no need to perform an operation such as removing the culture solution 2 using a pipette or the like prior to the spectroscopic inspection, and there is an advantage that the possibility of contamination can be reduced and the operation can be simplified.

In the present embodiment, the case where the stage 5 is fixed in an inclined state has been described. Instead of this, as shown in FIG. Further, the light source 6 and the sensor 7 disposed so as to face each other in the vertical direction may be configured to be integrally swung by a predetermined angle as shown in FIG. By doing so, the culture vessel 4 is placed in a horizontal state on the stage 5 arranged in a horizontal state as shown in FIG. 2A, and the stage 5, the light source 6 and the sensor 7 are placed. Just by rocking, the sample 3 can be analyzed spectroscopically by removing the culture solution 2 as in FIG.
Further, with such a configuration, the sample 3 having no culture solution can be inspected with the stage 5 placed in a horizontal state as shown in FIG. 2 (a). There is an advantage.

  In the present embodiment, the culture vessel 4 containing the culture solution 2 and the sample 3 is illustrated as having a substantially circular bottom surface 4a. Instead, as shown in FIGS. The shape of the bottom surface 4a is preferably a shape having a different area across a straight line passing through the center of gravity, for example, a triangular shape. That is, in the example shown in FIG. 3, the area relationship between the two regions R1 and R2 (shaded portions) across the straight line Q passing through the center of gravity P is R1 <R2.

  In this way, as shown in FIG. 4, when the culture vessel 4 is tilted so that the region R2 on the side with the larger area of the bottom surface 4a is on the lower side, the volume below the center of gravity is larger. Therefore, the liquid level of the culture solution can be moved below the sample at a small angle. As a result, the sample can be exposed from the culture solution without increasing the tilt angle of the culture vessel 4, and spectroscopic inspection can be performed without being affected by the culture solution. By making the inclination angle small, it is possible to prevent the sample from shifting in the culture vessel. In addition, it is not limited to a triangle, You may employ | adopt other arbitrary shapes.

  Further, the culture vessel 4 having such a configuration is not limited to one having one recess 4b for accommodating the culture medium 2 and the sample 3, and a plurality of them are arranged vertically and horizontally as shown in FIG. You may adopt what is. Also in this case, as described above, each recess may have, for example, a triangular bottom surface 4a.

  In addition, as shown in FIG. 6, the recess 4b of the culture vessel 4 is adjacent to the culture part A where the sample 3 is immersed in the culture solution 2 and the bottom surface (culture surface) 4a of the culture part A. The observation part B having the observation surface 4c inclined and the culture solution storage part C arranged on the opposite side of the culture part A across the observation part B may be adopted.

In this way, as shown in FIG. 6A, the bottom surface 4a of the culture part A is disposed substantially horizontally, the culture solution 2 is stored, and the sample 3 is cultured, and then the spectroscopic inspection is performed. In this case, as shown in FIG. 6 (b), the culture vessel 4 is tilted so that the culture part A is on the upper side and the culture medium recovery part C is on the lower side, thereby being stored in the culture part A. The culture medium which has been flowing is allowed to flow into the culture medium recovery section C, and the sample 3 that has grown by adhering to the bottom surface 4a can be gradually moved to the horizontal observation surface 4c. In this state, by irradiating the near-infrared light L upward in the vertical direction, the spectroscopic inspection of the sample can be performed without being affected by the culture solution.
According to such a culture vessel 4, a commercially available spectroscopic inspection device that irradiates the near infrared light L upward in the vertical direction can be used.

  Further, as shown in FIG. 7, the recess 4b of the culture vessel 4 includes a culture part A in which the sample 3 is immersed in the culture medium and the inclined surface 4d on the bottom surface (culture surface) 4a of the culture part A. As a structure comprising an observation part B having an observation surface 4c adjacent to each other with the observation surface 4c substantially parallel to the bottom surface 4a, and a culture solution storage part C disposed on the opposite side of the culture part A across the observation part B Also good. Between the observation part B and the culture solution recovery part C, a partition wall 4e made of a mesh or the like that allows the culture solution 2 to flow and prohibits the passage of the sample 3 is provided.

  By doing in this way, as shown in FIG. 7A, the bottom surface 4a of the culture part A is arranged substantially horizontally, the culture solution 2 is stored, the sample 3 is cultured, and then the spectroscopic inspection is performed. In this case, as shown in FIG. 7 (b), the culture vessel 4 is tilted so that the culture part A is on the upper side and the culture medium recovery part C is on the lower side, thereby being stored in the culture part A. The culture medium which has been flowing is poured into the culture medium recovery section C, and the sample 3 which has grown by adhering to the bottom surface 4a is captured by the partition wall 4e.

Thereafter, by returning the culture vessel 4 until the observation surface 4c becomes substantially horizontal, the sample 3 can be placed on the observation surface 4c as shown in FIG. 7C. In this state, by irradiating the near-infrared light L upward in the vertical direction, the spectroscopic inspection of the sample can be performed without being affected by the culture solution.
Also with such a culture vessel 4, a commercially available spectroscopic inspection device that irradiates the near infrared light L upward in the vertical direction can be used.

  In the present embodiment, the sample 3 is irradiated with the near infrared light L emitted from the light source 6, and the near infrared light L transmitted through the sample 3 is detected by the sensor 7. Thus, the near infrared light L reflected from the sample 3 may be detected by the sensor 7 disposed on the same side as the light source 6.

It is a typical whole lineblock diagram showing the spectroscopic inspection device concerning one embodiment of the present invention. It is the 1st modification of the spectroscopic inspection device of Drawing 1, and is a figure showing the state where (a) the culture container was mounted horizontally, and (b) the state where the culture container was inclined, respectively. It is a top view which shows an example of the culture container which concerns on this embodiment suitable for applying to the spectroscopic inspection apparatus of FIG. FIG. 4 is a perspective view showing the culture container of FIG. 3, (a) a horizontal state and (b) an inclined state. FIG. 5 is a longitudinal sectional view showing a culture vessel having a plurality of recesses similar to FIG. 4, (a) a horizontal state and (b) an inclined state. It is a longitudinal cross-sectional view which is a modification of the culture container of FIG. 3, (a) is in a horizontal state, and (b) is in an inclined state. FIG. 4 is a longitudinal sectional view showing another modification of the culture container of FIG. 3, (a) a horizontal state and (b) an inclined state.

Explanation of symbols

A culture part B observation part C culture solution storage part L near infrared light P center of gravity Q straight line 1 spectroscopic inspection device 2 culture solution 3 sample 4 culture vessel (culture vessel for spectroscopic inspection)
4a Bottom (culture surface)
4b Recessed part 4c Observation surface 4d Inclined surface 4e Partition wall 5 Stage 6 Light source 7 Sensor 8 Spectrometer

Claims (8)

A stage for placing a culture vessel containing a sample together with the culture solution;
A light source that generates near-infrared light for irradiating the sample in the culture vessel placed on the stage;
A sensor for receiving near-infrared light irradiated from the light source and reflected or transmitted by the sample;
A spectroscopic measurement unit that measures near-infrared light received by the sensor and measures wavelength characteristics;
The spectroscopic inspection apparatus in which the stage is configured so that the culture container can be placed with an inclination relative to a horizontal direction.   The spectroscopic inspection apparatus according to claim 1, wherein the light source is arranged to irradiate near infrared light in a thickness direction of a sample in the culture vessel.   The spectroscopic inspection apparatus according to claim 1, wherein the light source, the sensor, and the stage are provided so as to be integrally swingable about a substantially horizontal axis. An exposure step of exposing the sample from the culture solution by tilting the culture vessel containing the sample with the culture solution;
An irradiation step of irradiating the sample exposed from the culture with near infrared light;
A light receiving step for receiving near-infrared light reflected or transmitted by the sample;
A spectroscopic inspection method comprising: a measuring step of measuring wavelength characteristics by spectrally analyzing received near-infrared light.   The spectroscopic inspection method according to claim 4, wherein the irradiation step irradiates near-infrared light along the optical axis along the thickness direction of the sample inclined with respect to the vertical direction by inclining the culture vessel. A culture container for spectroscopic inspection having a recess capable of accommodating a sample together with a culture solution,
The recess has a flat bottom surface on which the sample can be adhered and cultured;
A culture container for spectroscopic examination, wherein the bottom surface has a shape having different areas across a straight line passing through the center of gravity. A culture container for spectroscopic inspection having a recess capable of accommodating a sample together with a culture solution,
A culture part having a flat culture surface on which the concave portion can be cultured by adhering a sample; an observation part having a flat observation surface that is inclined with respect to the culture surface of the culture part and is adjacent to the culture surface; A culture solution storage unit that is disposed on the opposite side of the culture unit across the observation unit and has a volume for storing the culture solution stored in the culture unit when the observation surface is arranged substantially horizontally. A culture vessel for spectroscopic inspection. A culture container for spectroscopic inspection having a recess capable of accommodating a sample together with a culture solution,
The concave portion has a flat culture surface on which a sample can be adhered and can be cultured, and a flat surface that is adjacent to the culture surface and is substantially parallel to the culture surface across an inclined surface that is inclined with respect to the culture surface of the culture portion. An observation part having a proper observation surface, and the culture solution stored in the culture part when the inclined surface is arranged substantially horizontally, with the observation part being sandwiched between the culture part and the inclined part. A culture medium storage unit having a volume to store,
A culture container for spectroscopic examination, in which a partition wall is provided between the observation surface of the observation unit and the culture solution storage unit, and a partition wall that prohibits the passage of the sample and allows the culture solution to pass therethrough.

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