JPH026345Y2 - - Google Patents

Description

[Detailed description of the invention] [Industrial application field] This invention is a method for quantitatively dispensing cell culture fluid simultaneously and quantitatively from each well of a microplate, which is often used for small-scale cell culture. Regarding fractionators.

[Conventional technology]

Microplates are used for small-scale cell culture by having multiple wells drilled in multiple rows on the top surface, and each well is filled with cell culture medium. The operation of dispensing a certain amount of the old culture solution and filling it with new culture solution is repeated and periodically performed.

In such aliquoting of the culture medium to be replaced, it is necessary to prevent cross-contamination of the culture medium in each well, and for this reason conventionally a removable liquid reservoir tip is attached to the tip of the pipette. This is done by sucking up the liquid using vacuum suction and replacing the liquid reservoir chip.

[The problem that the idea aims to solve]

However, such methods carry the potential risk of possible cross-contamination, which is most objectionable in operations such as culturing. In addition, one suction can only suction within one well, and in microplates with a large number of wells, such as 96, it takes a long time and requires a lot of effort. For this reason, attempts have been made to arrange pipettes each equipped with a liquid reservoir tip in a line to perform simultaneous aspiration, but due to the large volume of the pipettes, installation space is limited, and the number of wells is limited to 96. In many cases such as the above, simultaneous suction was not possible, and suction could only be performed one row at a time.

[Means to solve the problem]

The present invention utilizes suction tips made of porous plastic in which many small holes and striations are formed, which have the property of sucking up solutions, etc. by capillary action, and are arranged in multiple rows. It is possible to simultaneously and quantitatively separate a large amount of culture fluid from within the well, thereby solving the above-mentioned drawbacks.

That is, the simultaneous quantitative sorting device for microplates of the present invention consists of a microplate with a plurality of wells formed on the upper surface to be filled with a cell culture solution, and a large number of small holes and lines that absorb the cell culture solution by capillary action. Suction tips made of porous plastic with stripes formed therein are disposed to face the wells and protrude from the bottom surface, respectively, and the suction tips are immersed in the cell culture medium in the wells to a predetermined depth. The device is characterized in that it includes a chip unit that is movable up and down.

〔Example〕

Hereinafter, one embodiment of the present invention will be described in detail with reference to the accompanying drawings.

FIG. 1 is a perspective view of a chip unit 1 used in an embodiment of the present invention, and FIG. 2 is a perspective view of the chip unit 1 in an attached state. In these figures, the base plate 2 is formed into a rectangular plate shape, and small holes 3, 3, . . . are formed in multiple rows in the vertical and horizontal directions on the lower surface. This base plate 2 is
It is made of plastic or metal, etc., and there are suction tips 4,
4... can be fitted respectively. The suction tips 4, 4, . . . are made of a material that absorbs liquid by capillary action. The material used is porous plastic with many small pores and lines (product name: ``Filterene'').
Kaken Co., Ltd.). This filterene has pores that communicate in a three-dimensional network inside the molded product, and the size and number of pores can be controlled by changing the molding conditions (heating temperature, heating time), mold material, heating method, etc. can.

Moreover, this filterene has the following characteristics.

(1) Good water wettability and excellent water absorption ability.

(2) It does not swell or lose strength when water is absorbed, and has excellent dimensional stability.

(3) It is lightweight and has sufficient strength for practical use.

(4) There is no cytotoxicity to cultured cells, and both the cell proliferation rate and antibody production ability of antibody-producing cells are good.

(5) The wicking of crystalloid and colloid solutions is extremely rapid, within 50 μ/2.5 seconds.

These suction tips 4, 4... all have the same shape,
They are molded to the same length and thickness, so that the amount of liquid sucked in by capillary action is the same.
In this embodiment, the lower end has a rounded cylindrical shape, but the shape is not limited as long as it satisfies the above conditions.

When in use, the suction tips 4, 4... are often fitted into the small holes 3, 3... on the lower surface of the base plate 2, disinfected and sterilized in advance, and attached integrally with the base plate 2. Attached to the member. The mounting member 10, as shown in FIG. , 2a... are inserted into the holes 11a, 11a... drilled at the four corners of the frame 11, the tip unit 1 is removably positioned on the frame 11, and thereby the suction tips 4, 4... Protrudes downward from inside the hollowed out part 13. The grip part 12 that supports the frame 11 supports the frame 11 by pinching it from above and below, and is movable up and down. Although not shown, this vertical movement is performed by a motor or cylinder installed in the base 14, and the amount of descent is adjusted so that the suction tips 4, 4... can quantitatively aspirate the culture solution. has been done. Due to the lowering of the gripping part 12, the tip unit 1 placed on the frame 11 is lowered, and the suction tips 4, 4 are lowered.
...is inserted into each well of the microplate.

As shown in FIG. 3, the microplate 5 is formed in the shape of a rectangular plastic plate, and a large number of wells 6, 6... are bored in multiple rows in the vertical and horizontal directions on the top surface. A predetermined culture solution is filled in each well 6,6. Since the suction chips 4, 4... attached to the lower surface of the base plate 1 are inserted into the wells 6, 6..., respectively,
The same arrangement and number of wells 6, 6, . . . are attached to the base plate 1.

Figure 4 shows the situation of preparative separation, and shows the attachment member 1.
0 (not shown) is lowered by a predetermined amount and the tip unit 1 on the attachment member 10 is also lowered, so that the suction tips 4, 4, . As shown by the solid lines, the cells are inserted into the respective wells 6, 6... of the microplate 5, immersed in the internal culture solution at the same time and at the same depth, and the culture solution is sucked up by capillary action to collect the culture solution. becomes possible. In this case, since the descending distance of the tip unit 1 is preset, as shown in FIG. Since the s d are all the same and only the culture solution in the immersed part (d part) is aspirated, the amount of culture solution sucked up by each suction tip is the same, and quantitative suction is not possible. It becomes possible.

As described above, according to this embodiment, all the suction tips are inserted into the wells at the same depth to suction the culture solution by utilizing the capillary phenomenon of the suction tips, so that the culture solution is sucked by each suction tip. Not only is the volume of the culture solution the same, making quantitative fractionation possible, but the initial purpose can be achieved despite the small size of the suction tips, making it possible to arrange a large number of suction tips. , which results in
The culture solution in all wells of the microplate can be aspirated at the same time. In addition, the entire device can be made smaller, the structure is simple, and there is no need for a pump for vacuum suction, making it easy to operate.
It becomes possible to significantly shorten the operation time. In addition, the suction tips perform completely independent fractionation, are sterilizable, and are made of inexpensive materials that can be discarded after each use, so there is no risk of cross-contamination.

The present invention is not limited to the above-mentioned embodiments, and various modifications are possible. For example, the suction tip and the base plate can be integrally molded instead of being made separately, and this makes it possible to attach the suction tip. The operation can be omitted. Alternatively, the tip unit may be moved manually without automation, and in this case, the tip unit may be mounted on a suitable mount or the like, and the amount of descent may be adjusted using a stopper.

[Effect of idea]

As explained in detail above, according to the present invention, with a simple structure, it is possible to simultaneously and quantitatively separate a large amount of the culture fluid in all the wells of a microplate, and it is possible to sterilize it. Moreover, it is possible to provide a quantitative aliquot for culture fluid without any cross-contamination, and it is possible to leave live cells at the bottom of the well, remove dead cells and other floating materials, and replace them with fresh culture medium. be.

[Brief explanation of the drawing]

Figure 1 is a perspective view showing a chip unit according to an embodiment of the present invention, Figure 2 is a perspective view showing how the chip unit is installed, Figure 3 is a perspective view of an example of a microplate, and Figure 4 is a state of separation. FIG. 5 is a partially enlarged sectional view. 1... Chip unit, 2... Base plate, 3...
...Small hole, 4...Suction tip, 5...Microplate, 6...Well.

Claims (1)

[Scope of utility model registration request] A microplate with a plurality of wells perforated on the top surface to be filled with cell culture solution, and a suction tip made of porous plastic with many small holes and striations that suck up the cell culture solution by capillary action. It is characterized by comprising a tip unit which is disposed opposite to the well and protrudes from the bottom surface thereof, and which is movable up and down so that the suction tip is immersed in the cell culture solution in the well to a predetermined depth. A simultaneous quantitative fractionator for microplates.