JP2569073B2 - Cell processing apparatus and cell processing method - Google Patents

Description

The present invention relates to an apparatus and a method for treating cells.

[Conventional technology]

Conventionally, as a chamber plate for holding cells, the Journal of the Japan Society of Precision Engineering Spring Conference (1987), p.8
As described at 45, there is a large number of compartments (chambers) arranged in a matrix on a Si wafer. As means for supplying cells to each chamber one by one, there is a means for sending cells together with a fluid using a flow cell as described in the same paper.

[Problems to be solved by the invention]

The above-mentioned prior art does not take any measures to eliminate the cells, and has a problem that a plurality of cells are supplied in a row to one chamber or dust or the like is mixed. An object of the present invention is to provide a cell processing apparatus and a cell processing method that can solve the above problem and supply only a specified number of cells in a chamber.

[Means for solving the problem]

The object is to provide a means for detecting the state of particles in a liquid containing cells, a plurality of chambers for accommodating single cells among the particles, and an exclusion hole for excluding particles other than the single cells. A cell processing apparatus characterized by having a plate with, and a step of detecting the state of particles in a liquid containing cells, and a single cell of the particles,
This is achieved by a cell processing method comprising: a step of accommodating a cell other than a single cell in a chamber provided in a plate; and a step of eliminating particles other than the single cells using an exclusion hole provided in the plate. .

[Action]

When the cells are fluidly supplied into the chamber, the cell position detecting means checks whether the cells are connected in a rosary or not, and determines whether or not the cells are necessary. By using this means, and a plate having a plurality of chambers for holding cells and a hole adjacent to the chambers for excluding cells, unnecessary cells can be eliminated, and one cell can be surely inserted into the chamber. Allows cells to be inserted.

〔Example〕

An embodiment of the present invention will be described with reference to FIG. The chamber plate 1 shown in the figure has chambers (C ₁ , C ₂ , C ₃ , C ₄ ,...) For holding the cells 2 and holes (P ₁ , P _2, ...) For excluding the cells. They are arranged in a matrix.
Desired cells are inserted into each chamber on the chamber plate 1 in the following procedure. That is, the cells 2 are supplied to the chamber 1 in a liquid flow in the nozzle 3, and are sucked and held on the chamber bottom by the suction nozzle 4.

Next, a procedure for securely inserting one cell into each chamber will be described in detail. The cells 2 flowing in the nozzle 3 scatter the laser beam 7 emitted from the laser light source 6.

The level of scattering of the laser beam generated at this time is detected by the photomultiplier 5, and it is determined whether the cell is a single cell, a plurality of cells connected to each other, or another foreign substance such as dust. In order to insert cells one by one into the chamber, it is necessary to discharge particles other than single cells through the holes (P ₁ , P ₂ ...). For this reason, when it is recognized that the particles are unnecessary, the particles are fed back to the drive unit 9 of the XY table through the processing of the control system 8 so that the nozzle 3 excludes the cells (P ₁ , P ₂ ). Then, the chamber plate 1 is moved so as to be located above the nearest clearance hole. As a result, of the supplied cells, only one independent cell that is not in a daisy chain can be inserted into the chamber.

Next, it will be shown that the elimination operation can be performed successfully by arranging chambers for isolating and holding cells in a matrix and appropriately providing holes for excluding the cells therein. FIG. 2 is a top view of the arrangement of chambers such that the vertical and horizontal pitches are P. The recess of the chamber has a truncated pyramid shape, and two slits are provided on the bottom surface for holding cells. Among them, the second row of A _22, A _24, ... for those of the even-numbered, placing holes to eliminate cells. Now, consider the operation of supplying one cell to A ₁₁ , A ₁₂ , and A ₁₃ among the chambers (A ₁ i, i = _{1 to} n) arranged in the first row. When the cells flow independently through the nozzle, the cells are sequentially supplied to A ₁₁ , A ₁₂ , and A _13. However, if it is determined that the beads are connected, the cells are eliminated at the exclusion hole of A ₂₂ . Cells of whether flowed nozzles independently judgment, the mutual distance 1 _C of cell flows one after another nozzle, is performed based on the relationship of the moving velocity V _P of the flow velocity V _C and Chillan bar plate. That is, if the following equation is satisfied when the distance between the nozzle position and the chamber to which the cell is to be supplied next is 1 _P , the cells may be determined to be flowing independently.

1 _P / V _P ≦ 1 _C / V _{C In the} above equation, the left side is the time during which the chamber can move, and the right side is the time during which the chamber must move to supply cells one by one.

Having thus described the A _11, A _12, the operation to put the cells one by one to Chiyanba of A _13, for also Chiyanba of _{A 14, A 15, A 16} , the same operation the A ₂₄ as exclusion hole in the cell Just do it. With respect to A ₂₁ , A ₂₂ ,... In the second row, the supply operation of the folded cells is performed in the order of A _2n , A _{2 (n−1)} . A ₃₁ , A ₃₂ ,
A ₃₃ ... Are operated in the same manner as in the first row so that A ₂₂ becomes an exclusion hole for the chambers of A ₃₁ , A ₃₂ , and A ₃₃ .

In this way, if the exclusion holes are provided at even-numbered positions from the (3m-1) th row (m = 1, 2,...), The operation of supplying cells one by one can be performed smoothly. .

In the examples, the even-numbered positions from the left of the (3m-1) -th line (m = 1, 2,...) Were used as the cell exclusion holes, such as A ₂₂ and A ₂₄ . In addition to the above-described embodiment, various methods of providing such exclusion holes are conceivable, such as providing one for each one. However, any of these can be easily analogized from the present invention.

In the embodiment, the bottom of the chamber is completely penetrated as the exclusion hole, but the chamber at a specific position is used as the exclusion hole as in the above-described embodiment to capture cells that are connected in several rosaries. It is also possible to have only the role. The cells guided to and trapped in such an exclusion hole may not be subjected to subsequent cell operations, or may be subjected to an elimination operation at an appropriate time.

〔The invention's effect〕

As is apparent from the above description, the operation of supplying cells one by one can be performed extremely easily in the cell holding chamber plate according to the present invention.

[Brief description of the drawings]

FIG. 1 is a cross-sectional view showing the concept of a mechanism for eliminating rosary-connected cells, and FIG. 2 is a front view showing an example of arrangement of exclusion holes suitable for eliminating rosary-connected cells. 1 ...... Chillan bar plate, 2 ...... cells, P _1, P ₂ ...... elimination hole, C _1, C ₂ ...... Chiyanba.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Hiroyuki Kobida 1-280 Higashi Koikebo, Kokubunji-shi, Tokyo Inside the Central Research Laboratory, Hitachi, Ltd. (72) Inventor Masatoshi Sakurano 390 Muramatsu, Shimizu-shi, Shizuoka Prefecture Hitachi Inside the Shimizu Factory

Claims (5)

(57) [Claims] 1. A means for detecting the state of particles in a liquid containing cells, a plurality of chambers for containing single cells among the particles, and an exclusion hole for excluding particles other than the single cells. And a plate provided with: 2. The cell processing apparatus according to claim 1, wherein said plurality of chambers and said exclusion holes are formed at a constant pitch in a matrix. 3. The cell processing apparatus according to claim 1, wherein a specific chamber among the plurality of chambers is used as an exclusion hole. 4. A step of detecting a state of particles in a liquid containing cells, a step of storing a single cell of the particles in a chamber provided on a plate, and a step of storing particles other than the single cell. Excluding using an exclusion hole provided in the plate. 5. The method according to claim 5, wherein the chamber containing the single cell is:
The cell treatment method according to claim 4, further comprising a step of containing a single cell different from the cell.