JP4424938B2 - Microgel cassette for electrophoresis - Google Patents

Description

  The present invention relates to a microgel cassette for electrophoresis, and more particularly to a microgel cassette capable of easily producing a very small microgel suitable for measurement by electrophoresis.

  A conventional gel for electrophoresis has a size of at least 7 to 8 cm square as described in FIG. 9 of JP-A No. 2001-165903 and the description thereof, and its production requires many steps and time. In addition, a large amount of gel material and a large amount of sample are required, and the migration time is long.

The applicant of the present application, as a proposal for solving these drawbacks in the above-mentioned Japanese Patent Application Laid-Open No. 2001-165903, reduces the amount of gel material and test sample used by producing a micro support plate for electrophoresis, that is, a microgel, Furthermore, although an invention that can significantly shorten the electrophoresis time has been disclosed, it is difficult to efficiently produce a microgel because it requires manual skill, and this has hindered the spread of electrophoresis using a microgel. There was a problem.
JP 2001-165903 A

  The problem to be solved by the present invention is that anyone can prepare a microgel for electrophoresis easily, quickly, accurately and without waste without requiring skill.

  In the present invention, a thin gel solution container sandwiched between two bank surfaces is provided on one inner surface of two rectangular transparent and colorless microplates to be joined, and is accommodated in the upper end of the container. A well provided with a comb-toothed comb portion for forming a well in the solidification process of the gel solution, and injecting a test sample by folding the comb-toothed comb portion after the gel solution contained in the containing portion is solidified This problem was solved by forming a microgel having an upper end at the upper end, thereby spreading the electrophoresis using the microgel.

  According to the microgel cassette of the present invention, a desired microgel can be quickly and accurately obtained without waste by simple operations such as easy mechanical assembly, filling of the gel solution, and folding of the comb portion after the gel is solidified. This produces the effect of being able to be produced and contributing to the widespread use of electrophoresis by microgel, which has many advantages.

  According to the second aspect of the present invention, there is an effect that the microgel cassette can be accurately assembled by a mechanical work that does not require any skill at all such as short pin and hole fitting.

  According to the third aspect of the present invention, there is an effect of providing a microgel capable of performing electrophoresis in which a gel preparation material is reduced and a migration time is shortened with a small amount of sample.

  According to invention of Claim 4, the effect which can prevent reliably that a gel solution oozes to the bank surface is produced.

  According to the fifth aspect of the invention, the bottom of the well of the microgel prepared by increasing the thickness of the bottom of the well into which the sample is injected is reinforced, and the effect of preventing damage such as mold deformation can be prevented.

  According to the invention described in claim 6, the adhesion force of the produced microgel to the microplate is strengthened, and the microgel sticks to the comb portion and peels off from the plate when the comb portion is removed. The effect which prevents is produced.

  According to the seventh aspect of the present invention, a microgel having a single long well and a microgel having a plurality of wells can be selected and prepared. The effect that it can be performed is produced.

  According to the eighth aspect of the present invention, there is an effect that it is possible to prevent the upper corner of the well from being hit by the comb teeth and being damaged by the rotation when the comb portion is removed.

  According to the invention described in claim 9, the fitted plates can be easily peeled off, and the microgel after migration can be taken out to the outside if necessary, or a fluorescent agent or other reagent can be added to the test sample. The effect which can be added is produced.

  According to the invention described in claim 10, since the fluorescence effect of the plate does not appear when the excitation light for fluorescing the test sample that has been made fluorescent by adding a fluorescent agent in advance or afterwards, This produces an effect that the result of electrophoresis can be clearly confirmed in photography.

  According to the invention of the eleventh aspect, there is an effect that the gel solution filling operation into the accommodating portion of the microgel cassette can be performed easily and quickly without waste.

  According to the twelfth aspect of the present invention, there is an effect that a gel solution can be filled without waste even in a small number of microgel cassettes.

In the present invention, two rectangular shaped low-fluorescence or non-fluorescence materials to be joined together are formed of colorless and transparent microplates on one inner surface between 20 and 30 mm in depth and depth of 0 to 30 mm. To provide a thin gel solution container having a size of 8 to 1 mm, and to form a single long well or a plurality of wells in the solidification process of the gel solution stored in the upper end of the container. The comb portion with the comb teeth is provided, and after the gel solution accommodated in the accommodation portion is solidified, the comb portion with the comb teeth is folded off to prepare a microgel having a well for injecting a test sample at the upper end portion. Thus, the two microplates can be joined closely by providing several short pins and holes for joining on the bank surface, and the inner surface of one microplate and the bank surface Ridges in between The inner surface of the other microplate facing the comb portion is provided with a concave groove that fits closely with the protrusion on the inner surface of the other microplate so as to prevent the gel solution from seeping out to the bank surface side. A step for thickening the bottom of the well of the microgel prepared in the step is provided, and a small containing portion of the same gel solution as the containing portion is provided at the corners of the bank surface on both sides of the comb portion to remove the comb portion. sometimes so as to prevent separation of the microgel, the comb teeth of the narrow preceding opposing upper surface upper corner side of the well to prevent damage to the upper corner of the comb portion of the well during the removing folding comb sloped In addition, the microgel cassette for electrophoresis is provided with an inclined surface for peeling between the plates formed on the surface and joined to one or several places outside the bank surface of one microplate.

FIG. 1 is a perspective view showing an example in which two plates of one microplate 1 and the other microplate 2 formed into a rectangular shape by injection molding or the like are formed by integral molding, and FIG. 2 is a plan view of the same. is there. During the microplate 1 as shown in FIG. 2 is adapted to decouple more as readily shown in FIG. 3 (a) to be folded connecting portion 3a of the both sides of the tentative connecting between them, and 3b Yes.

  As a material for forming the microplates 1 and 2, a colorless and transparent synthetic resin material (trade name: Mitsubishi Rayon VH-000, etc.) such as acrylic having low fluorescent or non-fluorescent characteristics is used.

  One microplate 1 is provided with a gel solution container 5 having a size of about 25 mm in length and width of about 0.8 mm in the middle between both banks 4 a and 4 b, and an upper end of the container 5. A comb portion 6 with long slot-shaped comb teeth 6a is provided for forming a well for injecting a test sample when the gel solution to be contained is solidified. The comb portion 6 can be easily removed from the folding recess line 7 formed on the back surface. Reference numeral 6b denotes an upper wall that defines the upper end of the accommodating portion 5, and reference numeral 6c denotes a folding tab. Small storage portions 7 a and 7 b of the same gel solution as the storage portion 5 are provided at corners of the bank surfaces 4 a and 4 b on both sides of the comb portion 6. Further, protrusions 8a and 8b are provided between the accommodating portion 5 and the both bank surfaces 4a and 4b, and holes 9a and 9b for joining to the other plate 2 are provided at two intermediate positions between the both bank surfaces 4a and 4b. Provided. Reference numerals 10a and 10b denote gap inclined surfaces provided on the outer corners of the both bank surfaces 4a and 4b, which form gaps into which nails and jigs enter when the other microplate 2 is joined.

  The other microplate 2 has the same shape as the one microplate, and is closely fitted to the short pins 11a and 11b and the protrusions 8a and 8b, which are fitted into the holes 9a and 9b of the microplate 1 on both sides of the inner surface, respectively. The concave portions 12a and 12b are provided, the step portion 13 is formed in the portion facing the comb portion 6, and the holes 9a and 9b of the microplate 1 and the short pins 11a and 11b are aligned and press-fitted. As shown in FIG. 3 (b), the microgel cassette 14 having the gel accommodating portion 5 in the middle can be accurately assembled simply by wearing.

  A prepared gel solution g such as acrylamide gel or agarose gel is injected from the upper or lower open part of the housing part 5 of the assembled microgel cassette 14. Since the gel solution g is not as viscous as water, as shown in FIG. 4, the gel solution g is filled without gaps in the housing portion 5, around the comb teeth 6 a, and in the small housing portions 7 a and 7 b. 8b and the concave grooves 12a and 12b are closely fitted to each other, so that the gel solution g can be prevented from exuding to the bank surfaces 4a and 4b.

Waiting for the filled gel solution g to solidify and then rotating and folding outward with the knob 6c of the comb portion 6 as shown in FIG. 5 (a), the comb portion 6 can be easily folded at the position of the dent line 7. Can be removed. Since the upper surface of the comb tooth 6a is formed by a tapered inclined surface 6d, even if the comb tooth 6a rotates to draw an arc based on the dent line 7 as shown in FIG. The top corner of the well 15 formed on the solidified microgel 16 can be removed without damaging the upper corner of the well 15 side . In addition, since the microgel 16 is extended to the small accommodating portions 7a and 7b provided on the bank surfaces 4a and 4b on both sides so as to be sandwiched between both plates 1 and 2, the microplate 2 is obtained by expanding the area of the microgel 16. Along with the strengthening of the adhesion force to the substrate, it is possible to surely prevent the manufactured microgel 16 from peeling off following the comb portion 6 to be removed.

  As shown in FIGS. 6 and 7, the microgel cassette 14 containing the microgel 16 produced as described above has a well 15 opened on an electrophoresis apparatus (not shown) for applying an electric field and a temperature gradient. Then, the test sample S is injected into the well 15 and used for the electrophoresis test. Since the microplate is formed of a low-fluorescence or non-fluorescence material, the microplate 1 is used when observing or photographing with the naked eye by applying excitation light to fluoresce the fluorescent agent previously added to the test sample S after electrophoresis. Since the fluorescence effect of itself does not act, observation and photographing of the electrophoretic image (band pattern) B can be performed accurately and precisely.

  Further, when it is desired to take out the test sample S after the electrophoresis test for use in other experiments or when a fluorescent agent is added after the electrophoresis, the gap inclined surfaces 10a and 10b provided at the outer corners of the microgel cassette 14 are used. It is possible to easily separate the two microplates 1 and 2 by putting a tip of a nail or an instrument into the plate.

  FIG. 8 shows another embodiment, which is the same as the previous example except that the comb tooth of the comb portion 6 is a comb tooth 6e having a multi-slot shape for forming a well divided into eight.

  FIGS. 9 to 10 show a gel caster 17 for quickly and efficiently filling the gel solution g when producing the microgel 16 in the microgel cassette 14 of the present invention. The gel caster 17 is sized to accommodate three stacked microgel cassettes 14 and has a storage portion 18 with one surface and an open top, and a fitting groove of the elastic gasket 20 at intervals around three sides on one side of the storage portion 18. A thick plate rectangular core plate 19 provided with 21, and a cover plate 23 that is thin and of the same shape as the core plate 19, is placed on one surface of the core plate 19 with an elastic gasket 20 interposed therebetween, and is fastened with screws 22 a and 22 b. As shown in FIG. 10, three microgel cassettes 14 are stacked and stored in the storage portion 18 with the comb portion 6 side up, and the cover plate 23 is applied from one side with the elastic gasket 20 in between. The three microgel cassettes 14 housed by tightening and fixing the screws 22a and 22b to be screwed in from the side are clamped without gaps, and the filling device is opened from the upper opening. It is to fill at (not shown) until the gel solution reaches the upper wall 6b of the comb section 6 of the micro-gel cassettes 14. By doing so, the gel solution can be filled easily into a small microgel cassette 14 that is inconvenient to handle without leaking around. The gel caster 17 can be used for stacking five microgel cassettes 14 by deepening the storage portion 18. Also, 24 shown in FIG. 9 is a spacer created with the same shape and volume as the main body outline of the microgel cassette 14, and two or one microgel cassette 14 is filled with the gel solution by the gel caster 17 containing three sheets. Is used as an embedding instead of the microgel cassette 14. When producing two microgels, one spacer 24 is used. When one microgel is desired, two spacers 24 are used to produce the required number of microgels 16 without wasting gel solution. Will be able to.

  According to the microgel cassette for electrophoresis of the present invention, a microgel can be prepared easily and accurately by anyone without requiring any skill at all. In addition, the electrophoresis test using a microgel that can be performed efficiently by further shortening the electrophoresis time can be widely spread.

(Embodiment 1) Perspective view showing an example in which two microplates are separated and easily formed integrally Same as above, top view (A) is a perspective view showing two separated microplates in a state before joining, (b) is a perspective view of the joined state. FIG. 3A is an end view taken along line AA in FIG. 3, FIG. 3B is an end view taken along line BB, and FIG. 3C is an end view taken along line CC. Figure (A) is the fragmentary sectional view which shows the state which removes the comb part, (b) is the elements on larger scale of the process Plan view showing a state in which a test sample is injected into the well of the prepared microgel The top view which shows an example of the electrophoresis image (band pattern) of the test sample after the electrophoresis performed by attaching a temperature gradient to the left and right (Example 2) Perspective view showing an example of a multi-slot shape for producing a well in which comb teeth are divided into a plurality of pieces (Example 3) The perspective view which isolate | separates and shows each member of the gel caster for filling a gel solution with a microgel cassette The perspective view which shows the state which accommodated the micro gel cassette in the accommodating part, and was screwed and fixed similarly

Explanation of symbols

1 is one microplate 2 is the other microplate 3a and 3b are joint portions 4a and 4b are bank surfaces 5 is a gel solution container 6 is a comb portion 6a is a comb tooth (long slot)
6b is an upper wall 6c is a knob for removing a fold 6d is a narrow inclined surface 6e is a comb tooth (multi-slot)
7a and 7b are small accommodating portions 8a and 8b are protrusions 9a and 9b are bonding holes 10a and 10b are gap inclined surfaces 11a and 11b are short pins for bonding 12a and 12b are concave grooves 13 and a paragraph portion 14 is micro Gel cassette 15 is well 16 is micro gel 17 is gel caster 18 is storage part 19 is core plate 20 is elastic gasket 21 is fitting groove 22a, 22b is screw 23 is cover plate 24 is spacer g is gel solution S is test sample B is an electrophoretic image (band pattern)

Claims (10)

A container for storing a thin gel solution sandwiched between bank surfaces on both sides is provided on one inner surface of two rectangular transparent and colorless microplates to be joined, and the gel solution stored in the upper end of the container A comb-toothed comb portion is provided for forming a well in the solidification process, and the well into which the test sample is injected by removing the comb-toothed comb portion after the gel solution contained in the containing portion is solidified A microgel cassette for electrophoresis characterized by forming a microgel included in the above. 2. The microgel cassette for electrophoresis according to claim 1, wherein the two microplates are closely joined by fitting several short pins and holes provided on the bank surface. The microgel cassette for electrophoresis according to claim 1 or 2, wherein the accommodating portion for accommodating the gel solution has a length and width of 20 to 30 mm and a depth of 0.8 to 1 mm. A protrusion is provided between the inner surface of one microplate and the bank surface, and a concave groove that closely fits the protrusion is provided on the inner surface of the other microplate so that the gel exudes to the bank surface side. The microgel cassette for electrophoresis according to any one of claims 1 to 3, wherein this is prevented. The microgel cassette for electrophoresis according to any one of claims 1 to 4, wherein a paragraph portion is provided on the inner surface of the other microplate facing the comb portion to reinforce the bottom of the gel well. The gel container according to any one of claims 1 to 5, wherein a small container portion of the same gel solution as the container portion is provided at the corners of the bank surface on both sides of the comb portion to prevent the gel from peeling when the comb portion is removed. Microgel cassette for electrophoresis. The microgel cassette for electrophoresis according to any one of claims 1 to 6, wherein the comb tooth has a long slot shape that forms a single long well or a multi-slot shape that forms a plurality of wells. 8. The comb teeth of claim 1 to 7, wherein an upper surface facing the upper corner side of the well is formed with a tapered inclined surface so as not to damage the upper corner side of the well when the comb portion is removed. The microgel cassette for electrophoresis according to any one of the above. The microgel cassette for electrophoresis according to any one of claims 1 to 8, further comprising a gap inclined surface for peeling between the plates joined at one or several locations outside the bank surface of one microplate. The microgel cassette for electrophoresis according to any one of claims 1 to 9, wherein the microplate is formed of a low fluorescent or non-fluorescent material.

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