JPH05501050A - Sample material transfer device - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Sample material transfer device The present invention is an apparatus for removing a sample of material from a body of semi-rigid material, such as an agar culture medium. , more specifically, especially when large numbers of colonies with different characteristics or grown on the same culture plate are used. The present invention relates to a device for transferring cells from one culture plate to another when cells are placed in the culture plate.

In microbiology, colonies of cells with various characteristics are grown on agar plates or culture plates. grown in a layer of water and nutrients. In order to perform experiments on cells, these cells are It is necessary to transport the agar from within the agar culture medium layer to the surface. This is done using This is immersed in an agar culture medium at the cell colony, where the cell colony is preferably Attach it to the bottle set. Next, the bottle set is placed on top of the agar culture layer on the other culture plate. Touches the surface softly. The intention of doing this is to place the cells on the surface of the second culture plate. The goal is to allow them to be deposited and grow into new settlements.

This method is labor intensive, inefficient and unreliable. These shortcomings should be replicated. Molecular biological applications such as large arrays of cultured colonies or cultured platelets. In one example, each settlement is a human game. There are different parts of the genome.

In such applications, it is important to ensure that the replica village corresponds exactly to the conditions of its original village. It is extremely important that one village is not contaminated by neighboring villages. It is essential. If such transfer were to be carried out by hand, the operation would be extremely difficult. It is labor intensive, time consuming, and error prone.

According to the invention, a sample of material is removed from a body of semi-rigid medium, such as an agar culture medium. A sampling device with a diameter per square centimeter of the sampling surface. It includes a large number of sampling protrusions or sampling surfaces with a certain arrangement density, and these The protrusions are formed integrally with the base, and the base or these protrusions are interposed between them. Inserting these protrusions into the body of the medium while holding them tightly in spaced relation. a sample collection device adapted to receive said medium from said body; is provided.

The present invention also provides a transfer device for transferring microbial colonies from between semi-rigid culture media. , a respective holder of a source medium and a source medium and a colony within said source medium. and a transfer device for collecting the sample and transferring the sample to the source medium, The apparatus is capable of harvesting substantially greater than the density of colonies per unit area of said source medium. It has a large number of sampling protrusions with a density per unit area of the surface, and these protrusions are connected to the substrate. The base is integrally formed with these protrusions separated by M distances between them. This is done by inserting these protrusions into the body of the medium while holding them tightly in relation. providing a transfer facility adapted to receive the medium from a body of the invention.

The present invention further provides for replicating arrays of cell colonies using a device or equipment according to the present invention. The method comprises: placing the device on a culture plate containing the cleavage array to be replicated; a step of allowing cells from each colony to adhere to protrusions of the device; at least some of the cells or the new culture plate. and transferring the culture plate to the surface of the culture plate.

Conveniently, these projections are cross-sectioned or square.

These projections and the base are machined from a block of metal or molded from tick material.

As will become clear from the following description, the present invention provides a method for extracting cell colonies from other media. To provide an effective device for transferring media.

The invention will be explained in more detail with reference to the following description and the accompanying drawings, which Of these drawings, FIG. 1 is a corner perspective view of a partially completed device according to the present invention. the law of nature, Figure 2 shows the completed device shown in Figure 1 (the dimensions of the protrusions are shown in Figure 2 for clarity). Schematic diagram of automated equipment for cell colony transfer, including (note exaggeration) It shows.

FIG. 1 shows a device according to the invention ft1 in the form of one block, which The block consists of two equally spaced parallel orthogonal walls that create a set of parallel walls. A set of grooves 2a, 2b are cut. These sets of grooves are arranged in a perpendicular array. It forms a number of rigid pillars or protrusions 3, with the tops of these protrusions being flat. It matches the plane.

These grooves are usually on the order of 1-2fflff+ deep, and the protrusions are on the order of 1-2fflff+. It is of a height of -. These grooves are a fraction of a millimeter wide.

Dozens of grooves are cut per centimeter, making it a fraction of a square millimeter. It forms protrusions, and many protrusions, even tiny ones, are formed per square centimeter. There is.

The dimensions and density of these protrusions are determined by the actual manufacturing technology used to cut the grooves. It is regulated by the fineness that can be applied to metals such as aluminum. In the case of devices made by It can be formed as follows. Machined block concavity (impres) sion) to produce relatively inexpensive equipment made of plastic wherever possible. It can be used as a mold. These things can be done cheaply enough cleaning of the equipment between uses, which is required if the equipment is to be disposed of after use and reused. The need for cleaning and sterilization may be avoided.

In one suitable construction, the pins are spaced 1.2 mm apart (i.e., between the center of the bottle and and the center of the milled groove is set to a pitch of 2 millimeters). normal 22a Using the transfer device dimensions of n square, this would be a 200x200 bin array, all ie 40,000 bottles per device.

The upper and lower limits of pin spacing are 0.5 and 1, respectively, in normal applications. ．． It can be made 5mm.

Usually the colonies are about 1 mm in diameter, and sometimes smaller. small In order to obtain a pick-up rate of about 80% for villages, a pin spacing of 0.5 mm is required. Any suitable and, in any case, optimum spacing can be determined by simple experimentation.

Usually, on one culture plate, there are 10 cells each consisting of a single cell with a diameter of about 1 mr+. There are approximately 000 villages. Using a sampling device with an array of 2°0 x 200 protrusions. When the protrusions are pushed into the agar medium and then pulled out, there are usually two or more The three protrusions collect cells from each colony each time during the collection process. some protrusions that are close to and therefore have the ability or potential to pick up cells from (e.g. 3 or 4) are always present. Large and dense arrangement of protrusions The degree also ensures that cells from different colonies can be kept separate from each other.

The stiffness and arrangement density of the projections 3 are such that the projections 3 maintain their normal spacing during operation. to avoid contamination of the sample (such as could occur if the projections were bent). This has a significant effect in that it is desirable for accurate transportation of settlements.

In the embodiment of the drawing, the groove is cut by a square tool and has a rectangular cross section. , so the protrusion is flattened at the top. However, different shapes for cutting grooves are required. to make a pointed or other shaped protrusion by using a shaped tool Can be done.

Other methods of forming grooves can also be used, such as chemical etching.

The method of using this device is as follows. Arrange cell colonies or agar culture medium on culture plate. grown in a layer of water and nutrients. Each colony is a cell with specific characteristics each colony contains a different arbitrary part of the human genome, for example. , colonies are identified by their position on the culture plate.

In order to be able to perform tests or experiments on cell colonies, the cell colonies must be It needs to be on the surface of the culture medium, rather than being buried within it.

It is also common to have multiple copies of colony arrays for multiple testing and control purposes. It is advantageous if To make such copies, a device according to the invention is used.

Thus, in use, the device transfers the source medium (and then the cell colony) with an even pressure. (desired source media), and then The protrusions retain material from this source medium when the tray is withdrawn.

The device is then moved towards the destination medium, e.g. a new agar plate, and plunged into it. A village is ``planted'' by imposing an origin. Then the source medium is be reproduced for a short period of time, after which further copies may be retrieved be done. Cells deposited on the surface of the destination culture plate multiply and form new colonies. .

In some applications, there is no need to wait for this regeneration to occur. The reason is that Each protrusion takes out a large number of cells per colony, allowing for collection/transfer/deposition of a large number of cells. The process is carried out successively to place a large number of colonies on various destination boards or different areas of a large board. This is because it creates the same number of copies.

The device according to the invention allows rapid and reliable replication of arrays of colonies. protrusion A large placement density ensures that the replicas have the same distribution as the original colony, making it easier for the replicas to Allows for easy identification.

The device 1 is attached to a handle or the like for manual use. alternative Typically, the device includes a source medium for picking up the sample by the device. a sample collection movement for the sample, a transport movement to move the device to the destination medium, and a The deposition movement to the destination medium for depositing the liquid is carried out under the control of the controller 3. an automatic transfer device 2 (e.g. an XYZ axis controller) which can be programmed to The attached attachment may be provided with a device.

The picking surface of this device need not be rectangular or square as shown; Alternatively, if it is desired to pick up colonies from a circular agar culture plate, for example, a circular It can be done.

Also, the picking surface of this device does not need to be flat.

In one alternative, the picking surface is a gently convex surface (e.g. part of a cylindrical surface). used to perform a gradual transfer movement or a pick-up action. I can stay. This means that if the agar culture medium is Picking up settlements from an area will be incomplete, but it is possible to use a convex picking surface. This is an educational benefit because it can be improved by Similarly, this pick-up device It may be cylindrical with a bottle provided on its circumference, preferably a holder. The holder is biased by a gently acting return spring toward the reference orientation within the holder. mounted so that it can be rotated. This is done by rolling this device over the surface of the agar culture medium. can be used to pick up colonies from long, narrow areas of agar culture plates. After this transfer operation, the spring will pick up the device in its reference orientation in the holder. , thus setting the village sample to the same layout as the original sample (similarly (by the rolling action of) allows the deposition operation to be carried out.

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Claims (14)

[Claims] 1. A sample in which a sample of material is removed from a body of semi-rigid medium, such as an agar medium. A sampling device that has a large placement density per square centimeter of sample collection surface. a sample collection surface provided with a large number of collection protrusions, and these protrusions are formed integrally with the base body, and this base body connects these protrusions with a distance between them. Inserting these protrusions into the body of the medium while holding them firmly in place; a sample collection device adapted to receive said medium from said body; 2. The protrusions are formed by an array of closely spaced vertical pairs of grooves on the sampling surface. Apparatus as defined in claim 1. 3. Claim 1 or 2, wherein the free end of the protrusion conforms to a flat plane. Equipment described in Section. 4. Claims wherein said protrusion and said base are machined from a block of metal. Apparatus according to paragraphs 1, 2 or 3 of the subsection. 5. The protrusion and the base body are molded from a plastic material. Apparatus according to scope 1, 2 or 3. 6. Any of claims 1 to 5, wherein the protrusion has a square cross section. The device according to item 1. 7. Transfer equipment for transferring microbial colonies between semi-rigid culture media, holder of each of the source and destination media and the colonies in the source media and the a transfer device for transferring the sample of the sample to the destination medium; A unit area of the sampling surface that is substantially greater than the density of settlements per unit area of the source medium. It has a large number of collection protrusions with a certain density, and these protrusions are formed integrally with the base body. This substrate holds these protrusions firmly in spaced relationship between them. the body of the medium by inserting these protrusions into the body of the medium. Transfer equipment adapted to receive storage media. 8. The protrusions are formed by an array of closely spaced vertical pairs of grooves on the sampling surface. 8. The equipment according to claim 7, which is made of a single surface. 9. Claim 7 or 8, wherein the free end of the protrusion conforms to a flat plane. Equipment described in Section. 10. The claim 1, wherein the protrusion and the base are machined from a block of metal. Equipment according to scope 7, 8 or 9. 11. Claim in which the protrusion and the base are molded from a plastic material. Equipment described in item 7, item 8 or item 9 within the scope of. 12. Claims 7 to 11, wherein the protrusion has a square cross section. Equipment described in any one of the above. 13. a sampling movement relative to the source medium; Transfer movement between the holders of the destination medium and sample deposition on said destination medium. an automated transfer machine operable to drive said collection device by a landing motion; The equipment according to any one of claims 7 to 12, comprising a structure. 14. The device according to any one of claims 1 to 6 or Cell colony arrangement using the equipment described in any one of the scope items 7 to 13. A method of duplicating a column, the method comprising: Place the device on a culture plate containing the array to be replicated and collect the samples from each colony. allowing cells to adhere to the protrusions of the device; and placing the device in a new culture plate. at least some of the cells in contact with the surface are transferred to the surface of the new culture plate. A method comprising the steps of: