JP5253752B2 - Photosynthesis sample measuring container and container holder - Google Patents

Description

  The present invention relates to a photosynthetic sample measuring container used to store a photosynthetic sample and to measure the stored photosynthetic sample, and a container holder for holding the photosynthetic sample measuring container.

When measuring the absorbance of a photosynthetic sample, for example, algae or plant cells having a photosynthetic function, with a measuring device, a quartz measuring cell is usually used. In such measurement, it is necessary to transfer a photosynthetic sample previously cultured in a culture vessel to a measurement cell for measurement. On the other hand, according to the sample cell described in Patent Document 1, since culture and measurement can be used in a single container, there is no need to transfer the cultured sample to another cell.
JP-A-7-147968

  However, in the sample cell described in Cited Document 1, although the time of culture and the time of measurement can be shared by a single cell, if priority is given to the culture efficiency, the measurement accuracy tends to decrease. Tends to decrease. Further, in the sample cell described in Cited Document 1, the culture efficiency is not necessarily high because a sufficient interface between the sample and the atmosphere cannot be ensured.

  The object of the present invention is to provide a photosynthetic sample container and a container holder that can improve the culture efficiency and also improve the measurement accuracy.

In the photosynthetic sample measurement container used for storing the photosynthetic sample and measuring the stored photosynthetic sample, the present invention includes a first wall portion that becomes a bottom during culture, and a first wall portion that is substantially orthogonal to the first wall portion . And connected to the second wall portion so as to be substantially orthogonal to each other, and connected to the second wall portion which becomes the bottom at the time of measurement , and substantially perpendicular to the first and second wall portions, and is incident on the photosynthetic sample. A third wall portion having a light incident window for transmitting incident light, and being substantially orthogonal to the first and second wall portions and connected to face the third wall portion, facing the light incident window. A fourth wall having a light exit window that transmits incident light that has propagated through the photosynthetic sample at a position where the photosynthesis sample is transmitted, and an inlet for injecting the photosynthetic sample, the first wall being more than the second wall. The area of the part is larger.

  According to the present invention, at the time of culture, the first wall portion having a larger area than the second wall portion becomes the bottom, and the photosynthetic sample spreads thinly along the first wall portion. Therefore, the surface area where the photosynthetic sample comes into contact with the atmosphere at the time of culture increases, and the gas exchange of the culture solution is promoted to improve the culture efficiency. Further, at the time of measurement, since the second wall portion having a smaller area than the first wall portion becomes the bottom, the photosynthetic sample becomes bulky on the second wall portion in the container. Therefore, in the photosynthetic sample measurement container, the volume of the culture solution sufficient for the absorbance measurement through the light entrance window and the light exit window can be secured, so that the measurement accuracy is improved. As a result, the culture efficiency can be improved and the measurement accuracy can be improved.

  Furthermore, it is preferable to provide a breathable lid that covers the inlet. Since the air-permeable lid can maintain air permeability while preventing the entry of various germs and the like, it is possible to further improve the culture efficiency.

  It is preferable that the photosynthetic sample measurement container is provided with a holding unit for holding the photosynthetic sample inside the container. When the photosynthetic sample is held inside the container, the photosynthetic sample can be kept in a predetermined position, which is effective for automation when a solution such as a diluted solution or a culture solution is injected into the photosynthetic sample.

  The holding part is preferably provided inside the first wall part. Since the first wall portion becomes the bottom during culture, it is possible to reliably mix the photosynthetic sample and a solution such as a diluent to be injected into the container.

  Furthermore, it is preferable that the photosynthetic sample is held in advance in the holding unit. By collecting the photosynthetic samples in the holding unit, drying due to dispersion of the photosynthetic samples can be prevented, and deterioration of the photosynthetic samples can be prevented.

  Further, it is preferable that the injection port is provided in a wall portion facing the first wall portion, and the holding portion is disposed at a position facing the injection port. Since the solution injected from the injection port with a dispenser or the like is brought into direct contact with the photosynthetic sample and mixed, the mixing loss between the photosynthetic sample and the solution can be reduced, and the mixing is performed quickly.

  Furthermore, it is preferable that the holding part has a solution receiving part that receives the solution to be injected and a guide part that guides the flow direction of the solution dropped onto the solution receiving part. The photosynthetic sample and the solution can be efficiently mixed while guiding the solution in a predetermined direction.

  Furthermore, it is preferable that the holding portion has a lid portion in which a large number of holes are formed. In addition to preventing the photosynthetic sample from flowing out from the holding portion into the container, for example, when the solution is injected from the inlet, the photosynthetic sample in the holding portion is prevented from scattering.

  It is preferable that the holding part has a higher thermal conductivity than the other part of the wall part forming the photosynthetic sample container. With this configuration, the temperature control of the photosynthetic sample during culture is efficiently performed via the heat conducting member. When the photosynthetic sample is held in the holding unit in a frozen state, the photosynthetic sample can be efficiently frozen and thawed.

  According to the present invention, it is possible to provide a photosynthetic sample measurement container and a container holder that improve the culture efficiency and also improve the measurement accuracy.

  Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.

(First embodiment)
1 and 2 are perspective views of a photosynthetic sample measurement container that accommodates a photosynthetic sample. As shown in FIG. 1, the photosynthetic sample C in the photosynthetic sample measuring container 1A is cultured in a state diluted with a diluent W, for example, and irradiated with excitation light L1 after a predetermined time (see FIG. 2). ). Delayed luminescence (also referred to as “delayed fluorescence”) L2 is generated from the photosynthetic sample C by irradiation with the excitation light L1, and the state of the photosynthetic sample can be evaluated by measuring the temporal change of the delayed luminescence L2. First, the evaluation of environmental factors using this evaluation will be described.

(Evaluation of environmental factors)
When the photosynthetic sample C such as algae is irradiated with the excitation light L1, the photosynthetic sample C performs photosynthetic metabolism and grows cells. In photosynthetic metabolism, light energy absorbed by a photosynthetic pigment is transmitted by a plurality of chemical reactions and converted into energy necessary for cell growth. In the process, reactions that serve as energy sources for delayed luminescence such as enzyme generation, photo-chemical energy conversion, and CO ₂ absorption sequentially occur. Photons are generated (emitted) at different timings from these reactions, and the sum of the emitted light is measured as a weak delayed emission L2 of the entire photosynthetic sample.

  When harmful environmental factors act on the photosynthetic sample, the intracellular metabolism changes, and the temporal change in the amount of delayed luminescence is different from that when no environmental factors act. Furthermore, the influence on delayed light emission differs for each environmental factor. That is, the influence of each environmental factor on the photosynthetic sample C can be evaluated by comparing the time-dependent changes in delayed light emission.

(Container)
A storage container (photosynthesis sample measurement container) 1A for storing a photosynthetic sample is made of a transparent resin and has a substantially rectangular parallelepiped shape. The container 1A includes a flat lower wall portion 3 (first wall portion) that becomes the bottom when the photosynthetic sample C is cultured (see FIG. 1), that is, when the photosynthesis sample C is laid on a mounting table such as a shelf or a desk. And a flat upper wall portion 5 facing the portion 3. The lower wall portion 3 and the upper wall portion 5 are both transparent, and the lower wall portion 3 and the upper wall portion 5 are transmission windows 4 and 6 through which excitation light and delayed emission are transmitted over the entire surface.

  Furthermore, the container 1A includes a first side wall 7 (second wall), a second side wall 9, and a third side wall that surround the four surfaces so as to connect the lower wall 3 and the upper wall 5 to each other. It has the part 11 (3rd wall part) and the 4th side wall part 13 (4th wall part). The first side wall 7 and the second side wall 9 are opposed to each other, and the third side wall 11 and the fourth side wall 13 are arranged to face each other. The first side wall portion 7 is flat and becomes the bottom during measurement, that is, when it is set on a sample mounting table of an optical measurement device (not shown). Further, the lower wall portion 3 preferably has an area five times or more larger than that of the first side wall portion 7.

  As shown in FIGS. 1 and 3, a holding portion 15 protruding in an annular shape is provided inside the lower wall portion 3. A concentrated photosynthesis sample C (see FIG. 3) is stored in the holding unit 15. If the photosynthetic sample C is provided in the holding unit 15 in advance, it is possible for the measurer who has acquired the storage container 1A with the photosynthetic sample C to reduce the trouble of putting the photosynthesis sample C into the storage container. Furthermore, by providing the photosynthetic sample C whose growth conditions and quantity have been standardized in advance, variation in the concentration of the photosynthetic sample can be suppressed, so that the measurement accuracy can be improved.

  A circular inlet 17 is formed in the upper wall portion 5 facing the lower wall portion 3. Further, the injection port 17 is formed at a position facing the holding unit 15 so as to be directly above the holding unit 15. The standardized photosynthetic sample C is introduced from the inlet 17 into the holding unit 15 and stored frozen. Further, when the photosynthetic sample C is cultured, as shown in FIG. 4, a diluted solution W composed of an aqueous solution for adjusting pH, a culture solution containing nutrient salts, and the like is supplied from the injection port 17 by the pipette tip 18 or the like. It is dripped.

  As described above, the standardized photosynthesis sample C is held at a predetermined position by the holding unit 15. Therefore, when the diluent W is injected into the photosynthetic sample C, the troublesomeness of searching for the position of the photosynthetic sample C and aiming at each injection is reduced, and the diluent W can be injected with the same operation. As a result, it is also effective when automating the dilution of the photosynthetic sample C. Furthermore, since the standardized photosynthetic sample C is held in a narrow region in the holding unit 15, it is possible to prevent the photosynthetic sample from being dispersed and dried in the storage cell 1A. Deterioration can be prevented. In particular, by having the holding unit 15, it becomes easy to hold the standardized photosynthetic sample C in advance, and the efficiency of culture and measurement is improved.

  Further, when the standardized photosynthetic sample C is dispensed into the holding unit 15 and stored, it is preferable to store it frozen in order to suppress deterioration of the photosynthetic sample C. Further, when the photosynthetic sample C is cultured, it is necessary to thaw the photosynthetic sample C. Since the photosynthetic sample C is held in the holding unit 15, the photosynthetic sample C can be efficiently frozen and thawed by setting a cooling or heating point in the holding unit 15.

  Further, the injection port 17 is directly above the holding unit 15 during culture (see FIGS. 3 and 4). Therefore, when the photosynthetic sample C is diluted with the diluent W, the photosynthetic sample C is mixed in direct contact with the dropped diluent W, so that the loss of the diluent W can be reduced and the mixing is performed. This can be done quickly.

  As shown in FIG. 1, after diluting the photosynthetic sample C, a circular air-permeable lid (lid member) 21 that shields the injection port 17 is attached to the upper wall portion 5. By shielding the injection port 17 with the cover member 21 having air permeability, the air permeability can be maintained while preventing entry of germs and the like. As the air-permeable lid member, a membrane or a filter is preferably used. The lid member preferably has water repellency.

  The third side wall portion 11 is provided with an incident window 23 through which the incident light L3 irradiated to the photosynthetic sample C is incident, and the fourth side wall portion 13 has an emission window 24 that emits the emitted light L4. Is provided. The entrance window 23 and the exit window 24 are arranged to face each other. Further, the absorbance of the photosynthetic sample C is measured through the entrance window 23 and the exit window 24. Therefore, the entrance window 23 and the exit window 24 are the bottoms at the time of measurement rather than the second side wall 9 side. 1 side wall portion 7 (second wall portion) side, that is, in the vicinity of the first side wall portion 7 (second wall portion) that becomes the bottom during measurement. The incident light L3 is light for measuring the absorbance, and a special process (mirror finish or the like) is performed on the portion through which the incident light passes to obtain transparency, flatness, and parallelism. Since performing such a special process over the entire surface of the accommodation cell 1A causes an increase in cost, the entrance window 23 and the exit window 24 are subjected to a special process corresponding to the position where the transmitted light L3 is irradiated. And are provided.

(Containment cell holder)
As shown in FIG. 5, the storage container 1 </ b> A is inserted into a case-shaped storage container holder (container holder) 2 </ b> A and set on a sample mounting table (not shown) of the optical measurement device. The storage container holder 2A has a rectangular parallelepiped shape corresponding to the storage container 1A, and an upper surface is opened to form an insertion port 25 of the storage container 1A. The storage container 1A is inserted into the insertion port 25 in a state where the first side wall portion 7 is positioned at the bottom, and the first side wall portion 7 abuts against the bottom surface 26 of the storage container holder 2A. In the storage container holder 2A, a light incident port 27 that exposes the incident window 23 of the storage container 1A and a light output port 29 that exposes the output window 24 of the storage container 1A are formed. Furthermore, the container holder 2A is formed with an excitation light incident port 31 that exposes a part of the lower wall 3 of the container 1A and a delayed light emission exit 33 that exposes a part of the upper wall 5. ing.

  The excitation light L1 irradiated toward the storage container 1A passes through the excitation light incident port 31 and the lower wall portion 3 and reaches the photosynthetic sample C to excite the photosynthetic sample C. The delayed light emission L2 generated by the irradiation of the excitation light L1 is measured through the delayed light emission outlet 33. The incident light L3 irradiated toward the storage container 1A enters the storage container 1A via the light incident port 27 and the incident window 23, and the outgoing light L4 transmitted through the photosynthetic sample C is emitted from the output window 24 and The light is emitted through the light emission port 29. The absorbance is measured by measuring the emitted light L4.

  At the time of culture (see FIG. 1), the container 1A has the lower wall 3 having a larger area than the first side wall 7 at the bottom, and the photosynthetic sample C diluted with the diluent W It spreads thinly along. Therefore, the surface area where the photosynthetic sample C comes into contact with air at the time of culturing is increased, gas exchange accompanying the growth of the photosynthetic sample C is promoted, and culture efficiency is improved. At the time of measurement (see FIG. 2), the storage container 1A is inserted into the storage container holder 2A, and the first side wall part 7 having a smaller area than the lower wall part 3 is the bottom. Therefore, the photosynthetic sample C diluted with the diluent W becomes bulky and thick on the first side wall portion 7. Therefore, the measurement accuracy of the delayed light emission L2 and the transmitted light L4 is increased, and the measurement efficiency is improved. As a result, the efficiency at the time of culture can be improved and the efficiency at the time of measurement can also be improved.

  Further, as described above, the storage container 1A is inserted into the insertion port 25 of the storage container holder 2A and held in the storage container holder 2A. Since the storage container holder 2A includes the excitation light incident port 31, the delayed light emission output port 33, the light incident port 27, and the light output port 29, the storage container 1A can be stored in a state where the photosynthesis sample C can be measured. The container holder 2A is held in a stable state.

(Second Embodiment)
A second embodiment of the present invention will be described with reference to FIGS. FIG. 6 is a cross-sectional view of the container according to the second embodiment. 7 is a cross-sectional view taken along line VII-VII in FIG. 6, and FIG. 8 is a cross-sectional view taken along line VIII-VIII in FIG. In addition, about the element and member same as 1 A of storage containers which concern on 1st Embodiment, the same code | symbol is described and description is abbreviate | omitted.

  A holding portion 37 that holds the standardized photosynthesis sample C is formed integrally with the lower wall portion 35 inside the lower wall portion 35 of the storage container 1B. The holding portion 37 includes an elliptical annular peripheral wall portion 38 and a concave curved portion 39 formed inside the peripheral wall portion 38. The longitudinal direction of the peripheral wall portion 38 coincides with the longitudinal direction of the lower wall portion 35. Further, the curved portion 39 includes a solution receiving portion 41 for receiving the diluent and a guide portion 43 for guiding the flow of the diluent W. The solution receiving part 41 is disposed on the side close to the second side wall part 9 and is deepest, and the guide part 43 is gently inclined and disposed on the side close to the first side wall part 7, The side far from the solution receiver 41 is shallower than the near side. The guide part 43 guides the dilution liquid W received by the solution receiving part 41 to the first side wall part 7 side. The standardized photosynthetic sample C is placed on the solution receiving portion 41, and the diluted solution W dropped on the solution receiving portion 41 is guided in the flow direction by the guide portion 43 after contacting the photosynthetic sample C. While flowing, it is mixed efficiently.

(Third embodiment)
A third embodiment of the present invention will be described with reference to FIG. FIG. 9 is a cross-sectional view of the container 1C according to the third embodiment. In addition, about the element and member same as 1 A of storage containers which concern on 1st Embodiment, the same code | symbol is described and description is abbreviate | omitted.

  The holding portion 15 of the lower wall portion 3 of the storage container 1 </ b> C is provided with a dome-shaped lid portion 45 that covers the inside of the holding portion 15. A large number of holes 47 are formed in the lid portion 45. Covering the holding unit 15 with the lid 45 prevents the photosynthetic sample from flowing out of the holding unit 15 and, for example, the photosynthesis sample C in the holding unit 15 scatters when the diluent W is injected from the injection port. To prevent.

(Fourth embodiment)
A fourth embodiment of the present invention will be described with reference to FIG. FIG. 10 is a cross-sectional view of the container 1D according to the fourth embodiment. In addition, about the element and member same as 1 A of storage containers which concern on 1st Embodiment, the same code | symbol is described and description is abbreviate | omitted.

  A cylindrical fitting port 53 is formed in the lower wall portion 51 of the storage container 1D, and a bottomed cylindrical holding portion 55 is fitted into the fitting port 53 and bonded thereto. The other part of the lower wall part 51 excluding the holding part 55 is made of resin, and the holding part 55 is formed of aluminum, copper, or the like having a higher heat conduction efficiency than the other parts forming the housing container 1D. . Since the holding portion 55 has a relatively high heat conduction efficiency in the lower wall portion 51, it becomes easy to effectively maintain the photosynthetic sample C at an appropriate temperature. In particular, the temperature control during the cultivation of the photosynthetic sample C, Effective for cooling and heating during freezing and thawing.

(Fifth embodiment)
A fifth embodiment of the present invention will be described with reference to FIG. FIG. 11 is a perspective view of a storage container holder 2B according to the fifth embodiment. In addition, about the same element and member as 2 A of storage container holders which concern on 1st Embodiment, the same code | symbol is described and description is abbreviate | omitted.

  As shown in FIG. 11, the storage container holder 2B has a substantially rectangular parallelepiped shape corresponding to the storage container 1A, and an insertion port 57 into which the storage container 1A is inserted is formed on the upper surface. An excitation light incident port 59 through which excitation light passes is formed in the lower part of the container holder 2B, and the first side wall portion 7 of the accommodation cell 1A is applied to the edge 61 forming the excitation light incident port 59. Touch. Further, the container holder 2B is provided with a rectangular reflecting plate 63 on the wall facing the lower wall portion 3 of the container 1A. Further, the storage container holder 2B is provided with a light incident port 27, a light output port 29, and a delayed light emission output port 33, similarly to the storage container holder 2A according to the first embodiment. When the excitation light L1 is irradiated toward the accommodation cell 1A, the excitation light L1 passes through the excitation light entrance 59 and the first side wall portion 7 and reaches the photosynthetic sample. Delayed light emission L2 generated by irradiation with the excitation light L1 is emitted in all directions. Since the reflection plate 63 is disposed facing the delayed light emission outlet 33, the delayed light emission L <b> 2 reflected from the reflection plate 63 is also directed to the delayed light emission outlet 33. Therefore, it is possible to measure the weak light such as the delayed light emission L2 by effectively using the reflected delayed light emission L2, and the detection amount of the delayed light emission is increased, so that the measurement accuracy is improved.

(Sixth embodiment)
A sixth embodiment of the present invention will be described with reference to FIG. FIG. 12 is a perspective view of a storage container holder 2C according to the fifth embodiment. In addition, about the same element and member as 2 A of storage container holders which concern on 1st Embodiment, the same code | symbol is described and description is abbreviate | omitted.

  The container holder 2C is made of resin, and the excitation light incident port 31, the light emission outlet 33, the light incident port 27, and the light emission port 29 are formed in the same manner as the container holding tool 2C according to the first embodiment. Yes. A flexible positioning piece (positioning portion) 67 for positioning the container 1A is formed on the rear wall portion 65 where the excitation light incident port 31 is formed. The positioning piece 67 protrudes from the upper part of the wall part 65, and the annular protrusion part 69 is formed inside the front-end | tip part. On the outside of the lower wall 3 of the storage container 1A, there is an annular recess 71 (see FIGS. 3 and 13) that forms the holding portion 15, and the projection 69 of the positioning piece 67 is fitted into the recess 71 to accommodate the storage container. Position 1A to prevent it from coming off. Accordingly, since the container 1A can be accurately placed at a predetermined position of the container holder 2C, the optical path for measurement can be determined with good reproducibility.

  The photosynthetic sample measurement container and the container holder according to the present invention are not limited to the above embodiments, and for example, a window that transmits excitation light, delayed light emission, incident light, and the like through one incident window provided in the photosynthesis sample measurement container. You may make it also combine.

  In addition, the holding unit does not have to face the injection port. For example, the holding unit may be provided at a corner in the storage container and a position where a pipette tip or the like inserted from the injection port can reach.

It is a perspective view which shows the storage container which concerns on 1st Embodiment of this invention, and shows the time of culture | cultivation. It is a perspective view which shows the storage container which concerns on 1st Embodiment, and shows the time of measurement. It is sectional drawing of the storage container which concerns on 1st Embodiment. It is a perspective view of the storage container holder which concerns on 1st Embodiment. It is sectional drawing which shows the state which has dripped the dilution liquid with the pipette tip. It is sectional drawing of the storage container which concerns on 2nd Embodiment. It is sectional drawing along the VII-VII line of FIG. It is sectional drawing along the VIII-VIII line of FIG. It is sectional drawing of the storage container which concerns on 3rd Embodiment. It is sectional drawing of the storage container which concerns on 4th Embodiment. It is a perspective view of the storage container holder which concerns on 5th Embodiment. It is a perspective view of the storage container holder which concerns on 6th Embodiment. The container holding tool which concerns on 6th Embodiment is shown, (a) is a side view which shows the state in the middle of inserting the container, (b) is a side view which shows the state which positioning of the container was completed It is.

Explanation of symbols

  1A, 1B, 1C, 1D ... container (photosynthesis sample measuring container), 2A, 2B, 2C ... container holder (container holder), 3, 35, 51 ... lower wall part (first wall part), 4 , 6 ... transmission window, 7 ... first side wall (second wall), 11 ... third side wall (third wall), 13 ... fourth side wall (fourth wall) , 15, 37, 55 ... holding part, 17 ... injection port, 21 ... membrane filter (lid part), 23 ... light entrance window, 24 ... light exit window, 57 ... insertion port, 27 ... light entrance port, 29 ... light Exit port 31, 59... Excitation light entrance port 33. Delayed light emission exit port 41. Solution receiving portion 43. Guide portion 45. Cover portion 47 47 hole 63 Reflector plate 67 Positioning piece (positioning) Part), C ... photosynthesis sample, W ... diluent.

Claims (9)

In the photosynthetic sample measurement container used for storing the photosynthetic sample and used to measure the stored photosynthetic sample,
A first wall that becomes the bottom during culture;
A second wall portion connected to the first wall portion so as to be substantially orthogonal, and serving as a bottom during measurement;
A third wall portion connected to the first and second wall portions so as to be substantially orthogonal to each other and having a light incident window that transmits incident light incident on the photosynthetic sample;
The incident light propagated through the photosynthetic sample is transmitted to a position opposed to the light incident window , being substantially orthogonal to the first and second wall portions and connected to face the third wall portion. A fourth wall having a light exit window;
An injection port for injecting the photosynthetic sample,
The photosynthetic sample measuring container, wherein the area of the first wall portion is larger than that of the second wall portion.   The photosynthetic sample measurement container according to claim 1, further comprising an air-permeable lid that covers the injection port.   The photosynthetic sample measuring container according to claim 1, wherein a holding unit for holding the photosynthetic sample is provided inside the photosynthetic sample measuring container.   The photosynthetic sample measurement container according to claim 3, wherein the holding portion is provided inside the first wall portion.   The photosynthetic sample measurement container according to claim 3 or 4, wherein the photosynthetic sample is held in the holding portion.   The said injection port is provided in the wall part facing the said 1st wall part, The said holding | maintenance part is arrange | positioned in the position facing the said injection port, The Claims 3-5 characterized by the above-mentioned. Photosynthesis sample measurement container.   The said holding | maintenance part has a solution receiving part which receives the solution injected, and a guide part which guides the flow direction of the said solution injected into the said solution receiving part. The photosynthetic sample measuring container according to one item.   The photosynthetic sample measurement container according to any one of claims 3 to 7, wherein the holding part has a lid part in which a large number of holes are formed.   The photosynthetic sample measurement container according to any one of claims 3 to 8, wherein the holding part has a higher thermal conductivity than the other part of the wall part forming the photosynthetic sample container.

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