Source: https://patents.google.com/patent/WO2003055973A1/en
Timestamp: 2018-12-11 04:01:27
Document Index: 420397930

Matched Legal Cases: ['art 1', 'art 2', 'art 2', 'art 2', 'art 2', 'art 2', 'art 2']

WO2003055973A1 - Reaction container and reaction container holding mechanism - Google Patents
Reaction container and reaction container holding mechanism Download PDF
WO2003055973A1
WO2003055973A1 PCT/JP2002/013639 JP0213639W WO03055973A1 WO 2003055973 A1 WO2003055973 A1 WO 2003055973A1 JP 0213639 W JP0213639 W JP 0213639W WO 03055973 A1 WO03055973 A1 WO 03055973A1
PCT/JP2002/013639
A reaction container (100) used for gene inspection, comprising at least one reaction liquid storage part for storing reaction liquid having upper side wells (152) opened in the upper surface of the reaction container (100), lower side wells (112) positioned on the underside of the upper side wells (152), and flow passages (114) communicating the lower side wells (112) with an outside space and reaction parts (132) disposed between the upper side wells (152) and the lower side walls (112), the reaction part (132) further comprising nucleic acid probes fixed to the frames of porous bodies, wherein the upper side wells (152) opened in the upper surface of the reaction container (100) allows the reaction parts (132) to be optically observed from the upper side of the reaction container (100).
The reaction vessel and the reaction vessel holding mechanism
The present invention relates to a reaction vessel used for genetic testing, in particular
About the Kokoroyo # 5 that will be suitably applied to genetic testing using a DNA microphone lower ray.
In recent years, the analysis of the genes of many plants, including a large number of organisms and rice, including non-we have been promoted.
Genetic testing, generally was done Ri by the electrophoresis method so far. In the electrophoresis method, and it requires a long time to inspection, limited to a small number the number of performed inspection at a time.
Recently, a novel test how have been developed using DNA chips or DNA microarrays were regularly arranged DNA into a semiconductor or the like. This new test method, it is possible to simultaneously inspect a plurality of genes.
However, this new test method requires a long time to test, it is difficult to obtain a good analysis result reproducibility.
Applicant No. 2 0 0 1 - proposed in 2 3 2 5 0 1 No., using 3D DNA microarray, a conveniently short time may be tested genes genetic testing apparatus and gene detection method ing.
In this genetic test, the reaction solution is placed a filter porous to the liquid containing portion was cut specifications by tertiary DNA microphone lower rays and sputum body (reaction section), pressurization and pressure reduction by the pump for example silicon Njipo pump which it passed through a repetitive reaction portion by. With good reproducibility nucleic acid in a short period of time This ensures that can be obtained.
JP 1 - 1 8 0 4 3 No. 5 and US 5, 5 5 2, 5 8 0 No. discloses a device for heating the microphone Ropure Bok.
These devices, while sealing the microphone Ropure Bok state, and are the single only the amplification reaction of nucleic acids, is not considered at all and this observing the nucleic acid reaction.
Primary object of the present invention is a Rukoto to provide a technology that enables optical observation while performing the reaction of genetic testing with temperature control. Therefore, secondary objective of the present invention is a child provide suitably applied may reaction vessel to the good Una genetic testing. Another secondary object of the present invention is a Rukoto to provide a reaction vessel holding mechanism capable of holding the reaction container easily detachable.
The present invention is, in part, Ri reactor der used for genetic testing, the reactor of the present invention, even least for for accommodating the reaction liquid has a single reaction liquid receiving unit, the reaction liquid container parts are an upper Ueru opened to the upper surface of the reaction vessel, and a lower Ueru located below the upper Ueru has a passage communicating the lower Ueru and the external space, the reaction vessel further includes an upper © Ell and includes a reaction section disposed between the lower © El, the reaction portion has a nucleic acid probe immobilized on the skeleton of the porous body, the upper having an opening on the upper surface of the reaction vessel © El is to allow optical observation of the reaction portion from above the reaction vessel. Preferred details, the total volume of the lower Ueru and the channel greater Ri by volume of the upper Ueru, lower Ueru and the flow path may accommodate the entire amount of the originally placed in the upper Ueru reaction.
The reaction vessel is preferred properly, the upper surface of the reaction vessel Ri Contact with groove further comprises extending upper Ueru or al, the groove is placed on the reaction vessel to prevent evaporation of the reaction solution by the cover glass connexion partially covered, flow through the air between the upper Ueru and the external space can be ensured.
The reaction vessel is preferred properly the reaction vessel groove further has a recess for limiting the mounting position of the cover glass which extends to the outside of the recess, the flow path favored properly flanking the reaction container opening further includes a re in g groove for receiving an 0-ring surrounding take end.
The reaction vessel is preferred properly is a liquid accommodating portion for holding the liquid-containing, a first groove the upper surface of the reaction vessel extending between the upper Ueru and liquid storage portion, the upper surface of the reaction volume liquid second groove and further comprising in per cent Li a extending from the accommodation unit, the second groove is partially covered by Kabaga laths placed on the reaction vessel to prevent evaporation of the reaction solution , the flow of air between the upper © El and the external space through the liquid storage portion is ensured.
The reaction vessel is preferred properly, and further has a projection formed on the bottom surface of the lower © el, projection limits the deformation of the reaction unit.
The present invention is, in part, Ri reaction vessel holding mechanism der for holding the above-mentioned reaction vessel used in the genetic testing, the reaction container holding mechanism of the present invention, a mounting table reaction vessel is placed, and anti-reaction container unit presses against which receiving portion, a mechanism pressing attaching press the receiving portion of the reaction vessel, and the first temperature adjusting mechanism for adjusting the temperature of the reaction vessel, the temperature of the atmosphere in the vicinity of the upper surface of the reaction vessel and a second temperature adjusting mechanism for adjusting.
Second temperature adjusting mechanism is favored properly, close to the Ku of the upper surface of the reaction vessel, the reaction vessel holding mechanism has a heating means arranged away from the reaction vessel, is preferred properly, above the reaction vessel further comprising a cover member disposed, the cover member Contact Li, cover member supports the heating means has a window that allows transmission of light, the window is located above the reaction portion of the reaction vessel, the reaction to allow optical observation of the reaction portion from above the container
Cover member, it is favored properly further comprises an optically transmissive Akirana member for closing a window spatially.
Figure 1 is an exploded perspective view of a reaction vessel according to the first embodiment of the present invention.
Figure 2 is an assembly perspective view of a reaction vessel shown in Figure 1. Figure 3 is Ru sectional view der the reaction vessel along the III-III line of FIG.
Figure 4 is a perspective view of a reaction vessel holding mechanism the reaction vessel of Figure 1 to 3 is assembled.
5, Ri side view der of the reaction vessel holding mechanism of FIG. 4, the cross section of the reaction container and the reaction vessel holding mechanism is partially shown.
Figure 6 is a cross-sectional perspective view of a reaction vessel according to a modification.
Figure 7 is a cross-sectional perspective view of a reaction vessel according to another modification. Figure 8 is an overall configuration of the reaction vessel holding mechanism is incorporated microscope schematically illustrates.
9, FIG. 1 0 is a perspective view of a reaction vessel of a second embodiment of the present invention is a perspective view of a reaction vessel holding mechanism the reaction vessel of Figure 9 assembled.
Figure 1 1 is 1 2 is another perspective view of a reaction container holding mechanism of FIG. 1 0, cross-sectional view of a main portion of the reaction vessel and 1 0 and 1 1 of the reaction vessel hold mechanism of FIG. 9 it is.
Hereinafter, the description for the embodiment of the present invention with reference to the drawings.
The reaction vessel 1 0 0, a reaction liquid containing portion at least, is favored properly has a plurality of reaction liquid containing portion, four reaction liquid storage unit in earthenware pots by illustrated example in Figure 1 and Figure 2. Each of the reaction liquid storage portion, as shown in FIG. 3, an upper © El 1 5 2 which is open on the upper surface of the reaction vessel 1 0 0, lower Ueru 1 located below the upper © el 1 5 2 1 2, and a lower © El 1 1 2 and the external space to that channel 1 1 4 contact.
In the following description, the reaction vessel 1 0 0, but are not limited to, proceed Mel was assumed to have four reaction liquid storage unit.
The reaction vessel 1 0 0 further has an upper Ueru 1 5 2 and the lower © El 1 1 2 reaction unit 1 disposed between the 3 2. Paraphrase lever, upper © El 1 5 2 and the lower Ueru 1 1 2 is partitioned by the reaction unit 1 3 2. The reaction unit 1 3 2, and a nucleic acid probe immobilized on the skeleton of the porous body.
The reaction vessel 1 0 0 has an upper Ueru 1 5 2 which is open on its upper face, such upper Ueru 1 5 2, the reaction vessel 1 0 reaction part 1 from above 0 3 2 optical thereby enabling the Do not observed.
The reaction vessel 1 0 0, to obtain the above-mentioned structure, as shown in FIG. 1, the base 1 1 0 and the upper plate 1 5 0 that constitutes the container body, the alumina porous body having a reaction portion 1 3 2 and a 1 3 0. Base 1 1 0 and the upper plate 1 5 0, for example, poly Kabone - Ri Bok made der, in between the alumina porous body〗 3 0 are secured together by any suitable technique such as screwing or bonding in here, using a packing member such as described above, each of the upper Ueru 1 5 2 with each of O-rings 1 5 5 each lower Ueru 1 1 every 2 0-ring 1 1 7 3 confidential It is favored arbitrariness to the state to ensure a gender. If so as to obtain for each © El airtightness Patsukin member, of course influence from the external space, preferred because it avoided the impact between Ueru arbitrariness.
The upper plate 1 5 0, the upper Ueru 1 5 2 and the tapered one shaped through-hole for defining a groove 1 5 extending the upper surface of the upper plate 1 5 0 from the through hole (upper © El 1 5 2) and a 4.
Base 1 1 0 includes a recess for defining a lower Ueru 1 1 2, recesses (lower side © El 1 1 2) and the flow path 1 1 4 to contact the external space. Yo Ri details, the base 1 1 0 has a flat side surface 1 1 8, the passage 1 1 4 extends from the bottom surface of the recess (lower © E Le 1 1 2), the base 1 1 0 We are terminating at side 1 1 8. Base 1 1 0 Furthermore, the side surface 1 1 8 (i.e. passage end surface), surrounds take an open end of the channel 1 1 4, the re Ngumizo 1 1 6 to accommodate the O-ring It has.
Base 1 1 0 made et al, in order to avoid damage ie cracks only and tear of the reaction section 1 3 2, for suppressing the deformation of the reaction element 1 3 2, recesses (lower © El 1 1 2) one formed on the bottom surface or may have a plurality of projections. Protrusion may be a pin-like shape, or may be a plate-like shape.
As described above, the reaction liquid containing portion, and an upper © El 1 5 2 and the lower Ueru 1 1 2 and the flow path 1 1 4 As shown in FIG. 3. The sum of the volume of the lower Ueru 1 1 2 and the flow path 1 1 4 is greater Ri by volume of the upper © El 1 5 2. Therefore, the lower © El 1 1 2 and the flow path 1 1 4 can accommodate the entire amount of the first upper © El 1 5 2 encased that reaction. Channel 1 1 4, in order to obtain a sufficient volume in order to suppress the evaporation of the extension so-called liquid lumps Ri may reaction solution may have a, preferably, the reaction vessel 1 0 0 have a flat upper surface and, it covers first glass 4 1 0 placed al is thereon. Cover glass 4 1 0 have covered the entire upper © El 1 5 2 is placed so as to cover the grooves 1 5 4 partially. Overall covering the upper Ueru 1 5 2 minimizes evaporation of the reaction mixture, partial covering of the groove 1 5 4 gives the passage of air between the upper © El 1 5 2 and the external space.
The good Ri preferred correct reaction vessel, in order to reliably ensure the circulation of air, the upper plate 1 5 0 further has a recess 1 5 6 for determining the placement position of the cover glass 4 1 0, recess 1 5 6 the Contact Li to have a flat bottom surface, the grooves 1 5 4 extends to the outside of the recess 1 5 6. Structural Toku徵 like this can reliably avoided from being covered the whole groove 1 5 4 by cover first glass 4 1 0.
The reaction vessel 1 0 0 may be subjected to various changes and modifications. For example, as shown in FIG. 6, the reaction vessel 1 0 0, the reaction liquid container portion for accommodating the reaction liquid (upper © El 1 5 2 and the lower © El 1 1 2 and the flow path 1 1 4) and added to the reaction section 1 3 2 may further have a liquid storage portion 1 6 2 for holding the liquid-containing. That is, the reaction vessel 1 0 0, the reaction liquid storage unit (the upper © El 1 5 2 and the lower Ueru 1 1 2 and the flow path 1 1 4), and the reaction section 1 3 2, a liquid storage portion 1 6 2 has a groove 1 6 4 to contact an upper © El 1 5 2 and the liquid yield volume portion 1 6 2, from the liquid storage portion 1 6 2 and extending buildings grooves 1 6 6.
Cover glass 4 1 0, the upper © El 1 5 2 and covers the whole of the liquid storage portion 1 6 2 and the groove 1 6 4 is placed so as to cover the grooves 1 6 6 partially. Of course, the reaction vessel may have a recess for limiting the mounting position of the cover glass 4 1 0 as shown in FIG.
Upper Ueru 1 5 2 via the liquid storage portion 1 6 2 containing water or buffer over is communicated with the outside space. That is, air distribution between the upper © El 1 5 2 and the external space through the liquid yield capacity portion 1 6 2 is ensured. Is contained in the liquid container portion 1 6 2 water or buffer one is the high Ri by the surrounding upper Ueru 1 5 2 humidified atmosphere. This ensures that, is further reduced evaporation of the reaction solution. The good UNA suppress evaporation by water and of buffers over is particularly suitable for the measurement of the measurement Ya high temperature with high accuracy. Also, in the future clinical inspection, to detect a number of the reaction solution by Ri automatically many Ueru conceivable. In such applications, highly accurate results are required, it Needing and Prospective strongly suppress the evaporation of the reaction solution. Suppressing the evaporation by the aforementioned water or buffer over are also suitable for this application.
The reaction vessel 1 0 0, as shown in FIG. 6, it is not necessary to have one of the liquid storage portion 1 6 2 for one reaction liquid storage unit. That is, as shown in FIG. 7, a reaction vessel 1 0 0 plurality of reaction liquid storage portion (upper © El 1 5 2 and the lower © El 1 1 2 and the flow path 1 1 4), a plurality of reaction sections 1 3 2, one of the liquid storage portion 1 7 2 may have a single groove 1 7 6 extending from the liquid storage portion 1 7 2. Through a plurality of reaction liquid containing portion of the upper Ueru 1 5 2 a plurality of grooves 1 7 4 through the communication with you re a liquid storage portion 1 7 2, the liquid storage portion 1 7 2 one groove 1 7 6 It has been in communication with the external space Te.
The reaction vessel 1 0 0 is removably attached to the reaction vessel holding Organization 2 0 0 shown in FIG. 4, for example.
The reaction vessel holding mechanism 2 0 0 includes a mounting table 2 1 0, and the receiving portion 2 3 0 provided in the mounting table 2 1 0, a pair of pressing mechanism 2 5 0 for fixing the reaction vessel 1 0 0 have. Receiving unit 2 3 0, the reaction vessel 1 0 0 flow passage end surface 1 1 8 has a abutting surfaces 2 3 2 that is devoted press. Pressing mechanism 2 5 0 against the receiving unit 2 3 0 The reaction vessel 1 0 0 holds reaction vessels 1 0 0.
Each press and with mechanism 2 5 0, for example, Ri machine 械的 mechanism der utilizing screws, the fixed part 2 5 2 fixed to the mounting table 2 1 0, lever 2 5 rotationally operated 4, and a slider 2 5 6 which moves in response to rotation operation of the lever portion 2 5 4. A pair of fixing portions 2 5 2, Ri by the width of the reaction vessel 1 0 0 has also been arranged in a slightly wide spacing also serves as a guide that defines the lateral position of the reaction vessel 1 0 0 mounted.
Receiving unit 2 3 0, as shown in FIG. 5 has a through hole 2 3 4 extending therethrough. One end of the through-holes 2 3 4 terminates in abutting application surface 2 3 2, the other end of the through-holes 2 3 4 are tubes 4 3 0 is attached. Tube 4 3 0, the through-holes 2 3 4 of the receiving part 2 3 0 which is connected a pump, not shown, for example, silicon Njipisu tons fluid includes an open end of the reaction vessel 1 0 0 of the channel 1 1 4 It is arranged at the same pitch.
The reaction vessel 1 0 0 toward the abutting surfaces 2 3 2 sides 1 1 8 That passage end face 1 1 8 receiving unit 2 3 0 channel 1 1 4 is terminated, the mounting table 2 1 It is placed on top of the 0. Lateral position of the reaction vessel 1 0 0, O connexion is substantially determined on the fixed part 2 5 2 of the pair of pressing mechanism 2 5 0, also the vertical position of the reaction vessel 1 0 0, the receiving part 2 3 0 and the slider 2 It is almost determined by the 5 6.
By the rotation of the lever 2 5 4 Li slider 2 5 6 is moved toward the receiving portion 2 3 0. Movement of the slider 2 5 6 Press and attached to the receiving part 2 3 0 The reaction vessel 1 0 0. The reaction vessel 1 0 0 through 0-ring 4 2 0, the receiving part 2 3 0 and Ri by the fixing of the reaction vessel 1 0 0 by such a reaction vessel holding mechanism 2 0 0 coupled hermetically, reaction flow path 1 1 4 of the container 1 0 0 is receiving unit 2 3 0 of the through-holes 2 3 4 and fluid communication. Accordingly, the flow path 1 1 4 of the reaction vessel 1 0 0 via the receiving unit 2 3 through holes 2 3 4 and Ju-loop 4 3 0 0, is pumped fluidly connected such Siri Njipisu tons .
The reaction vessel holding mechanism 2 0 0 Ni will this Yo may detachably hold the reaction vessel 1 0 0 easily.
The reaction vessel holding mechanism 2 0 0 has a part of the container a temperature regulating mechanism for controlling the temperature of the reaction vessel 1 0 0. Vessel temperature adjusting mechanism, for example, a heating means for heating the reaction vessel 1 0 0. Vessel temperature adjusting mechanism for the good Ri responsive fast temperature control, the reaction vessel 1 0 0 may further have a cooling hand stage for cooling. Cooling means is normally placed outside the reaction vessel hold mechanism 2 0 0.
Accordingly, the mounting table 2 1 0 is, for example, has a calling heats the heater one 2 1 2 in accordance with the current supply and a temperature sensor 2 1 4 for detecting the temperature. Alternatively, the mounting table 2 1 0 may have a circulation path for circulating the hot fluid, such as liquid or gas.
The reaction vessel holding mechanism 2 0 0, to avoid clouding onset raw cover glass 4 1 0, has a portion of the ambient temperature adjusting mechanism for adjusting the temperature of the atmosphere in the vicinity of the cover glass 4 1 0 and ambient temperature adjusting mechanism which includes a heating means for controlling the atmosphere in the vicinity of the cover glass 4 1 0. Therefore, the reaction vessel holding mechanism 2 0 0 4 and as 5 shown in reaction vessel 1 0 0 near the upper surface of the reaction vessel 1 0 0 disposed away from the plate-like cover member 2 7 has 0, plate-like cover member 2 7 0 is, for example, it has a human one coater 2 8 2 which generates heat in response to current supply, and a temperature sensor 2 8 4 for detecting the temperature. Alternatively, the plate-like cover member 2 7 0 may have a circulation path for circulating the hot fluid, such as liquid or gas.
Ambient temperature adjusting mechanism adjusts the temperature of the atmosphere in the cover first glass vicinity in order to prevent fogging of the cover glass. Atmosphere temperature adjusting mechanism may be Tamotere the temperature of the atmosphere of the cover glass near above the dew point. Dew point depends on the humidity of the atmosphere in the cover glass vicinity.
The reaction vessel 1 0 0, so as not to interfere with the movement of the reaction solution therein, unsealed. The reaction liquid in the reaction vessel 1 0 in 0 moves in response to vacuum and pressurization of the flow path by the pump. Movement of the reaction solution circulating air. Circulation of air, suppresses the occurrence of temperature gradients in the Ueru, to change the humidity of the atmosphere in the cover glass vicinity.
Accordingly, the temperature setting of the ambient temperature adjusting mechanism, like details, are set in consideration of the humidity of the atmosphere in the cover glass vicinity. However, in practice, it may be set to a temperature to suppress the fogging to such an extent that does not interfere with the optical observation.
For example, ambient temperature adjusting mechanism, the Kiri囲 air cover glass near may be adjusted to the set temperature by Ri higher temperature of the vessel temperature adjusting mechanism, also may be adjusted to the same temperature as that. The temperature setting of by the temperature setting of the vessel temperature adjustment mechanism Li high ambient temperature adjustment mechanism prevents Ri effectively that by the fogging of the cover glass. Temperature setting in the same ambient temperature adjusting mechanism and a temperature setting of the vessel temperature adjusting mechanism facilitates Ri by the control of the temperature of the reaction solution.
Temperature setting of the vessel temperature adjusting mechanism and the ambient temperature adjusting mechanism may be set to an appropriate temperature depending on the subject, such as the discovery. Setting temperature may especially but without limitation, the preferred properly is 3 5 ° C~ 9 5 ° C. Temperature within this range, (solved conformation) de Ne overnight over genes in the reaction mixture and to facilitate removing the bound gene from the probe. Temperature settings may Meniwa was the detection of genes, yo Ri favored properly is a 3 5 ° C~ 8 0 ° C.
The reaction vessel holding mechanism 2 0 0 also for fogging prevention of the cover glass may further have a mechanism for blowing air toward the cover glass.
Plate-like cover member 2 7 0, the operation of attaching and detaching the reaction vessel 1 0 0 To easily, is favored properly, 4 and mounting table 2 via the link 2 7 2 as shown in FIG. 5 It is provided movably relative to 1 0.
Plate cover first member 2 7 0 has a window 2 7 4 to allow optical observation of the reaction vessel from above. Window 2 7 4 plate cover 2 7 0, the preferred properly, near the upper surface of the reaction vessel, in order to make even Ri by the reaction vessel efficiently the hot atmosphere, is covered with an optically transmissive Akirana member ing. Such member is, for example, a glass, the thermal conductivity is high transparency correct preferred Ri good high material reaction vessel holding mechanism 2 0 0, as shown schematically in Figure 8, XY stage 3 1 through 0, the optical microscope 3 0 0 stearyl over di-, eg assembled to Z-stage 3 2 0. The reaction vessel holding mechanism reaction vessel 1 0 0 held by the 2 0 0 is optically observed Ri by upwardly by an optical microscope 3 0 0. Upper © El 1 5 2 of reaction vessel 1 0 0 As described above, since the open to the top, by optical microscopy 3 0 0, reaction occurring in the reaction section 1 3 2 can be observed from above.
For example, the reaction vessel holding mechanism 2 0 0 is moved to match the tie Mi ring of the XY stage 3 1 0 Thus optical observation, the reaction unit 1 3 2 reaction vessels 1 0 0 of the observation object of an optical microscope 3 0 0 disposed within the observation field of view, it is observation such as a fluorescent appearance by light microscopy 3 0 0.
The reaction vessel 1 0 0 Ru is appropriately mounted on the reaction vessel holding mechanism 2 0 0. Flow path 1 1 4 of the attached reaction vessels 1 0 0 via the receiving unit 2 3 tubes 4 3 0 connected thereto and the through-holes 2 3 4 0, is communicated with the pump. This ensures that the reaction mixture of reaction solution containing section of the reaction vessel 1 0 0 (upper © El 1 5 2 and the lower © El 1 1 2 and the flow path 1 1 4), across the air layer in the middle is the interior space and fluid communication of the pump.
Pumps, air present between the reaction solution was aspirated, thus may flow path 1 1 4 of the reaction vessel under reduced pressure. Depending on the vacuum, the reaction mixture passes through the reaction portion 1 3 2, moves from the upper Ueru 1 5 2 to the lower space (space of the lower © El 1 1 2 and the flow path 1 1 4).
The total volume of the lower space of the reaction liquid storage unit to cormorants I mentioned above (the lower Ueru 1 1 2 and the flow path 1 1 4), since the greater Ri by volume of the upper Ueru 1 5 2, first upper Ueru 1 5 2 put obtained reaction solution, the entire amount can be accommodated in the lower space. Reduced pressure in the lower space by the pump is carried out to the extent that the reaction mixture remains in the reaction vessel.
The pump may pressurize the flow path 1 1 4 of the reaction vessel. Depending on the pressure, the reaction liquid passes through the reaction portion〗 3 2, to move from the lower space (lower Ueru 1 1 2 and the flow path 1 1 4) to the upper © El 1 5 2.
Ri by the and this to Shi performed repeatedly reduced pressure and the pressure of the flow channel 1 1 4 of Ri reaction vessel by the pump, the reaction liquid passes through the reaction portion 1 3 2, the upper side Ueru 1 5 2 and the lower side alternate between the space (the space of the lower © El 1 1 2 and the flow path 1 1 4).
By a child in this manner repeatedly to the reaction solution is passed through the reaction portion 1 3 2, object observation, for example, nucleic acids can be detected in a relatively short time.
The reaction vessel 1 0 0 is, for example, a disposable, but is not limited thereto. That is, after a single use, although alumina multi Anashitsutai 1 3 0 is discarded, the base 1 1 0 and the upper plate 1 5 0 may be reused after cleaning.
The reaction vessel 1 1 0 0 Contact Li to have a single reaction mixture containing portion at least, the reaction liquid containing portion, as shown in FIG. 1 2, an opening on the upper surface of the reaction vessel 1 1 0 0 upper and Ueru 1 1 5 2, a lower Ueru 1 1 1 2 positioned below upper © El 1 1 5 2, lower ©: L Le 1 1〗 2 and the flow path 1 1 1 4 to contact the external space and have a door. The reaction vessel 1 1 0 0, the preferred properly plurality of reaction EkiOsamu volume unit, but are not limited to, for example, as shown in FIG. 9, it has four reaction liquid storage unit.
The reaction vessel 1 1 0 0 further has an upper Ueru 1 1 5 2 and the lower Ueru 1 1 1 arranged between the two reaction unit 1 1 3 2. The reaction unit 1 1 3 2, and a nucleic professional one-flop which is fixed to the skeleton of the porous body.
The reaction vessel 1 1 0 0 Propelled by one upper Ueru 1 1 5 2 which is open on its upper face, such upper © El 1 1 5 2, the reaction portion from above the reaction vessel 1 1 0 0 thereby enabling a 1 3 2 optical observation.
The reaction vessel 1 1 0 0, to obtain the above-mentioned structure, as shown in FIG. 1 2, and Ichisu 1 1 1 0 and the upper plate 1 1 5 0 base constituting the container body, the reaction unit 1 1 3 and a porous alumina body 1 1 3 0 with 2. Base 1 1 1 0 and the upper plate 1 1 5 0 Example Ri polycarbonate Bok manufactured der eg to in between the alumina porous body 1 1 3 0, any suitable technique such as screwing or bonding One and are fixed to each other.
The upper plate 1 1 5 0, te for defining an upper Ueru 1 1 5 2 - has a par-shaped through hole.
Base 1 1 1 0 comprises a recess for defining a lower © El 1 1 1 2, recesses (lower side Ueru 1 1 1 2) and the flow path 1 1 1 4 to contact the external space there. Yo Ri details, the base 1 1 1 0 has a flat side surface 1 1 1 8, the channel 1 1 1 4 extends from the bottom surface of the recess (lower © El 1 1 1 2), the base terminating in a 1 1 1 0 side 1 1 1 8. Base 1 1 1 0 further aspect 1 1 1 8 (i.e. passage end surface), the channel 1 1 1 4 surrounds take an open mouth end, 0 Li Ngumizo 1 for housing-ring and has a 1 1 6.
As described above, the reaction liquid containing portion, as shown in FIG. 1 2, and a upper © El 1 1 5 2 and the lower Ueru 1 1 1 2 and the flow path 1 1 1 4. The total volume of the lower Ueru 1 1 1 2 and the flow path 1 1 1 4 is greater Ri by volume of the upper Ueru 1 1 5 2. Therefore, the lower © El 1 1 1 2 and the flow path〗 1 1 4 can accommodate the entire amount of the first upper © El 1 1 5 2 put is the reaction solution. Flow path 1 1 1 4, in order to obtain a sufficient volume, which may have a Ri extension so-called liquid lumps.
The reaction vessel 1 1 0 0 In addition to the reaction section 1〗 3 2 reaction liquid receiving unit (upper © El 1 1 5 2 and the lower © El 1 1 1 2 and the flow path 1 1 1 4), a liquid further includes a liquid containing portion 1 1 6 2 for receiving and holding. Liquid storage portion 1 1 6 2 consists of a rectangular recess that opens to the upper surface of the reaction vessel 1 1 0 0. In other words, the upper plate 1 1 5 0 that has a recess defining a liquid accommodating portion 1 1 6 2.
The reaction vessel 1 1 0 0, 9, has the reaction liquid storage portion and the number of the liquid storage portion 1 1 6 2, the number of the liquid storage portion 1 1 6 2 is limited to rather than shall state, and are not capable of being appropriately changed, for example, the reaction vessel〗 1 0 0 but it may also have one of the liquid storage portion 1 1 6 2 for a plurality of reaction liquid storage unit.
Upper Ueru I 1 5 2 and the opening end of the liquid storage portion 1 1 6 2 that is situated anti reaction container vessel 1 1 0 0 uppermost plane 1 1 5 8 good Ri is low. In other words, the upper plate 1 1 5 0 has a recess 1 1 5 6 rectangular recessed from the uppermost plane 1 1 5 8, it upper © Ezore 1 1 5 2 and the liquid the housing portion 1 1 6 2 that surround. That is, is formed to the upper © El 1 1 5 recess defining a tapered through-hole and the liquid containing portion 1 1 6 2 defining 2 recesses of the rectangle 1 1 5 6 bottom of.
Et al is, the upper plate 1 1 5 0, the grooves 1 1 6 6 extending between the flow path end surface 1 1 1 of 8 recesses closest to 1 1 5 6 side and the channel termination surface 1 1 1 8 It has. Grooves 1 1 6 6, as described later, the anti-reaction container vessel 1 1 0 0 Te state odor held in the reaction vessel holding mechanism, an upper © El 1 1 5 2 and the reaction vessel 1 1 0 0 external space providing a flow path for the air which allows the circulation of air between.
The reaction vessel 1 1 0 0 reaction vessel holding mechanism 1 2 0 0 detachably holding the can, as shown in FIG. 1 0 and 1 1, the base 1 2 with a recess for accommodating the reaction vessel 1 1 0 0 1 0, a base 1 2 2 0 the base 1 2 1 0 is placed, a base 1 2 2 0 pair of frames which is fixed to 1 2 2 4, pivot Bok the frame 1 2 2 4 1 2 and arm 1 2 2 6 coupled via the 2 8, and an arm 1 2 2 6 cover member 1 2 7 0 fixed on. Base 1 2 1 0 includes a table 1 2 1 O a reaction vessel 1 1 0 0 is placed, the reaction vessel 1 1 0 0 and a 1 2 1 O b unit receives devoted Press there. Arm 1 2 2 6 obtained rotated by the axis Pivot 1 2 2 8, therefore, can be opened and closed with respect to the cover member 1 2 7 0 Habe one Interface 2 1 0.
The reaction vessel holding mechanism 1 2 0 0, further, the reaction vessel 1 1 0 0 in conjunction with the opening and closing operation of the cover member 1 2 7 0, the base 1 2 1 0 receiving unit 1 2 Press and pressed to 1 0 b of with which it has a mechanism 1 2 5 0. Press and with mechanism 1 2 5 0 has a sliding sul slider 1 2 5 2 by communicating motion to the opening and closing of the cover first member.
As the slider 1 2 5 2, as the cover member 1 2 7 0 is closed, closer to the reaction vessel 1 1 0 0 placed in the recess of the base 1 2 1 0, cover first member 1 2 7 0 is opened , base - Ru away from Interface 2 1 reaction vessel 1 1 0 0 placed in the recess of 0.
The cover member 1 2 7 0 is closed, in other words, in the state in which the force bars 1 2 7 0 is placed on the base 1 2 1 0, the slider 1 2 5 2 shown in FIG. 1 2 as the reaction vessel 1 1 0 0, receiving unit 1 2 1 0 b reaction vessel 1 1 0 0 and SQLDESC_BASE_TABLE_NAME This result and against the sides of is maintained.
Further, in a state in which the cover member 1 2 7 0 is opened, the slider one 1 2 5 2 is away from the reaction vessel 1 1 0 0, the base 1 2 1 reaction vessel 1 from the recess of 0 1 0 0 easily it can be removed to take in.
However, if the cleaning by supplying the external I Ri cleaning liquid line Nau after completion of the reaction, the slider 1 2 5 2 and the cover member 1 2 7 0, mechanisms move independently, i.e., the slider is opened closed cover mechanism is favored arbitrariness not linked to. Receiving unit 1 2 1 O b of the base 1 2 1 0 has a through-hole 1 2 3 4 across the side facing the reaction container 1 1 0 0 flow passage end surface 1 1 1 8, a reaction vessel 1 in a state where 1 0 0 is maintained in the reaction vessel holding mechanism 1 2 0 0, the reaction vessel 1 1 0 0 is connected to the base 1 2 1 0 and hermetically through a 0-ring 1 4 2 0, the reaction vessel 1 1 0 flow path 1 1 1 4 0 is contacted base 1 2 1 0 of the through-holes 1 2 3 4 fluidly. Accordingly, the flow path 1 1 1 4 of the reaction vessel 1 1 0 0, via the through-hole 1 2 3 4 and the tube 1 4 3 0 of the base 1 2 1 0, like the pump and fluidly Syringe piston It is connected.
The reaction vessel holding mechanism 1 2 0 0 has a part of the container a temperature regulating mechanism for controlling the temperature of the reaction vessel 1 1 0 0. Container temperature adjustment mechanism is, for example, have a heating means for heating the reaction vessel 1 1 0 0. Container temperature adjustment mechanism, for a more responsive fast temperature control, cooling means for cooling the reaction vessel 1 1 0 0, for example, may have a cooling fan further. Cooling means is usually arranged outside the reaction vessel holding mechanism 1 2 0 0.
Therefore, the base 1 2 1 0 is, for example, as shown in FIG. 1 2, therein, the heater one 1 2 1 2 for generating heat in response to current supply, for detecting the temperature of the heater 1 2 1 2 and a temperature sensor 1 2 1 4. Alternatively, the base 1 2 1 0, instead having a human one coater 1 2 1 2 may have a circulation path for circulating the hot fluid, such as liquid or gas.
The cover member 1 2 7 0, in the closed Ji was state, has a window 1 2 7 4 to enable upward or these reaction vessel 1 1 0 0 reaction unit 1 1 3 2 optical observation . A member 1 2 7 6 optically transparent covering the cover member 1 2 7 0 further window 1 2 7 4, the leaf spring 1 2 9 2 for fixing the transparent member 1 2 7 6, the leaf spring 1 2 9 2 and possess bonded and Gomushi sheet 1 2 9 4 such as shea Li Kongomu the attached Li on. The leaf spring 1 2 9 2 and Gomushi sheet 1 2 9 4 together correspond to the window 1 2 7 4 position is opened.
The transparent member 1 2 7 6, for example Ri Glass der, is favored properly, high transparency thermal conductivity of high material.
In the state in which the cover member 1 2 7 0 is closed, Gomushi sheet 1 2 9 4 contacts the upper surface of the reaction vessel 1 1 0 0, the reaction liquid in the reaction vessel 1 1 0 in 0 serves to suppress the evaporation . However, never upper Ueru〗 1 5 2 of reaction vessel 1 1 0 0 is sealed, air flow has been secured between the grooves 1 1 6 6 upper Ueru 1 1 5 2 and the external space, it Ri air move easily obtained in the reaction vessel 1 1 0 in the 0 to, therefore the reaction liquid in the reaction vessel 1 1 0 in 0 without moving is prevented, depending on the pressure and the reduced pressure of the pump It can be easily moved.
Go厶Shiichi Bok 1 2 9 4 does not need to necessarily pasted to the leaf spring 1 2 9 2, may be bonded to the reaction vessel 1 1 0 0.
In order to suppress the evaporation of the reaction solution, the properly preferred Ri good, water or buffer, First is placed in the liquid containing portion 1 1 6 2. Is contained in the liquid container portion 1 1 6 2 water or of buffers scratch, and the upper © El 1 1 5 2 of high humidity Ri by the atmosphere around, suppressing Ri effectively by evaporation of the reaction solution. Suppressing evaporation by water or buffer over is particularly suitable for the measurement with high accuracy of measurement Teiya hot. Further, it is expected in future clinical inspection 査等 be suitably applied to test for detecting a number of the reaction solution by Ri automatically many Ueru. The reaction vessel of this embodiment, not only the genetic testing may also be applied to the inspection of cells or proteins or antigens or antibodies. Such a test, there is a case when cells or proteins or antigens or antibodies gene by Redirecting a hearing and the viscosity of the reaction liquid is high. When this occurs, the reaction solution since it is difficult to move, or by increasing the groove 1 1 6 6.
The reaction vessel of this embodiment, not only the genetic testing may also be applied to the inspection of cells or proteins or antigens or antibodies. The cells and was I Park quality and antigen and antibody, have multi-be would denatured Ri by the heat o
In reaction vessel holding mechanism 1 2 0 0 of this embodiment, there is a space between the heater 1 2 8 2 and the leaf springs 1 2 9 2, this space is influenced heater 1 2 8 2 directly to the reaction solution which serves to prevent the give. As a result, Saiichi Bashu preparative influence of heat such as would indirectly is, degeneration of the heat by the cells and proteins and the antigen or antibody can be suppressed.
The reaction vessel holding mechanism 1 2 0 0, like the first embodiment, mounted to Z Stage 3 2 0 of the optical microscope 3 0 0 for fluorescence observation through the XY Stage 3 1 0, a genetic testing device the reaction vessel 1 1 0 0 held in the reaction vessel holding mechanism 1 2 0 0 constituting, as in the first embodiment, is optically observed Ri by upwardly by an optical microscope 3 0 0. Since the upper Ueru〗 1 5 2 of reaction vessel 1 1 0 0 As described above is open to the top, by the optical microscope 3 0 0, reactions occurring in the reaction unit 1 1 3 2 can be observed from above.
Further, the reaction liquid in the reaction vessel 1 1 0 in 0 is passed through repeatedly reactive unit 1 1 3 2 by pressurization and depressurization of the pump in communication with the flow channel 1 1 1 4, observation This ensures object, for example, the nucleic acid is detected in a short time relatively.
Previously, have been described particular embodiments of the present invention with reference to the accompanying drawings, the present invention is, without having to go to be limited to the embodiments described above may be variously modified without departing from the spirit thereof industrial Applicability
According to the present invention, suitably applied may reaction vessel gene inspection heritage using 3D DNA microphone lower ray is provided. The reaction vessel holding Organization is provided capable of holding the reaction vessel easily detachable.
1. A reaction vessel used for genetic testing, the reaction solution comprises a least one of the reaction liquid accommodating portion for accommodating the reaction liquid containing portion, and the upper side Ueru opened to the upper surface of the reaction vessel , a lower © El which is located below the upper © El, Propelled by one of a flow passage communicating the lower © El and the external space,
Upper Ueru and lower Ueru arranged reaction part further Bei Eteori during the reaction portion has a nucleic acid pro Ichipu fixed to the skeleton of the porous body,
Upper © El opened to the upper surface of the reaction vessel is to allow optical observation of the reaction portion from above the reaction vessel, the reaction vessel.
2. In the claims〗, the total volume of the lower © El and the channel greater Ri by volume of the upper Ueru, lower Ueru and the flow path accommodates a total amount of the initially placed in the upper © El reaction obtaining a reaction vessel.
3. In claim 1 or claim 2, Ri your upper surface of the reaction vessel has a groove extending from the upper © El Furthermore, the groove is mounting on the reaction vessel to prevent evaporation of the reaction solution partially covered by the force bar glass to be location, air circulation between the upper © El and the external space is secured, the reaction vessel.
4. The method of claim 3, further has a recess for limiting the mounting position of the cover glass, the grooves reaction volume extends to the outside of the recess 0
5. In claim 1 or claim 2, a first groove extending the liquid storage portion for hold accommodating a liquid, between the upper Ueru and liquid storage portion the upper surface of the reaction vessel, the reaction vessel and the top surface has further a second groove extending from the liquid storage portion, partly by a cover glass is placed on the reaction vessel for the second groove to prevent evaporation of the reaction solution covered by, the flow of air between the upper © El and the external space through the liquid storage portion is Ru is secured, anti J Kokoroyo o
6. In any one of claims 1 to 5, Ri Li in g groove der surrounding take an open end of the flow path located to the side of the anti-reaction container unit, re for receiving the 0-ring further has, the reaction vessel ring groove.
7. In any one of claims 1 to 6, further has a projection formed on the bottom surface of the lower © el, projection limits the deformation of the reaction portion, the reaction vessel.
8. A reaction container holding mechanism for holding the reaction vessel used for genetic testing, the reaction vessel comprises a single reaction liquid storage portion also less for accommodating the reaction liquid, the reaction liquid container parts are an upper © El opened to the upper surface of the reaction vessel, Ri and lower © El which is located below the upper © el, and a flow path communicating the lower © El and the external space Contact, reaction container is provided with an arrangement by reaction unit between the upper © El and lower Ueru addition, the reaction portion has a nucleic acid probe immobilized on the skeleton of the porous body, the upper surface of the reaction vessel the reaction vessel holding mechanism der for optical viewing of the reaction portion from above the reaction vessel Ri by the upper © Ell opened to maintain the reaction vessel as possible is,
A mounting table reaction vessel is placed, and the reaction vessel and press against which the receiving part,
A push to give the pressing mechanism receiving portion and the reaction vessel,
A first temperature adjusting mechanism for adjusting the temperature of the reaction vessel, and a second temperature adjusting mechanism for adjusting the temperature of the atmosphere in the vicinity of the upper surface of the reaction vessel, the reaction vessel holding mechanism.
9. The method of claim 8, the second temperature adjusting mechanism is close to the upper surface of the reaction container, and a heating means arranged apart from the reaction vessel, the reaction vessel holding mechanism.
1 0. The method of claim 9, further comprising a cover first member disposed above the reaction vessel, cover one member Ri Contact supports the heating means, cover first member has a window that allows transmission of light , the window is located above the reaction portion of the reaction vessel, allowing optical observation of the reaction portion from above the reaction vessel, the reaction vessel holding mechanism.
1 1. The method of claim 1 0, the cover member further an optically transparent member for closing a window spatially has, the reaction vessel holding mechanism.
PCT/JP2002/013639 2001-12-26 2002-12-26 Reaction container and reaction container holding mechanism WO2003055973A1 (en)
JP2001395140 2001-12-26
JP2001/395140 2001-12-26
AU2002359915A AU2002359915A1 (en) 2001-12-26 2002-12-26 Reaction container and reaction container holding mechanism
EP20020793413 EP1464695A1 (en) 2001-12-26 2002-12-26 Reaction vessel and reaction vessel holding mechanism
JP2003556494A JPWO2003055973A1 (en) 2001-12-26 2002-12-26 The reaction vessel and the reaction vessel holding mechanism
US10873950 US20040235148A1 (en) 2001-12-26 2004-06-22 Reaction vessel and reaction vessel holding mechanism
US10873950 Continuation US20040235148A1 (en) 2001-12-26 2004-06-22 Reaction vessel and reaction vessel holding mechanism
WO2003055973A1 true true WO2003055973A1 (en) 2003-07-10
ID=19188938
PCT/JP2002/013639 WO2003055973A1 (en) 2001-12-26 2002-12-26 Reaction container and reaction container holding mechanism
US (1) US20040235148A1 (en)
EP (1) EP1464695A1 (en)
JP (1) JPWO2003055973A1 (en)
WO (1) WO2003055973A1 (en)
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