JP2002122523A - Semi-automated solid sample preparation method and structure - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make a microscope sample of a higher quality for diagnosing cells with a higher reproducibility and in a shorter time from a cell suspension, especially a specimen containing a flat epitherial cell or the like by a method for trapping desired cells selectively on a filter from the cell suspension while removing foreign matters. SOLUTION: A desired cell sample without any foreign matter is prepared by catching the flat epitherial cell or the like and the desired cell simultaneously with separate filters. A suction pressure is kept constant by sucking a fixed outside air simultaneously during the continuous suction. A cell suction sample is kept from adhering to the inner wall of a funnel to improve the operability with a structure and a mechanism that facilitates loading or unloading of the filter and the funnel. Mechanical automation is accomplished for parts alone susceptible to variations due to manual work among various conditions necessary for the making of a quality sample and the parts not susceptible thereto and time consuming portions are covered by manual work thereby enabling shorter time treatment.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for preparing a cell specimen for cytodiagnosis in the field of clinical examination and a structure of each part necessary for the method.

[0002]

2. Description of the Related Art Filtration and collection of cells from a biological suspension containing cells has been conventionally performed. However, in the case of manual operation, it is not possible to control the strength of suction, and the judgment at the end point may vary, so that excessive suction may cause cells to sink into the filter and deform, or insufficient fixation may dilute the fixative. Therefore, it was not possible to make a reproducible sample with large fluctuation factors.

As a means for solving these problems, an automatic fixed specimen preparing apparatus and method are disclosed in JP-A-2000-146.
No. 782. According to this method, when filtering and collecting cells from a suspension containing target cells, particularly a liquid biological sample, collecting the target cells in a pure form and without denaturing the cells and fixing the cells on a filter. Can be done.

However, also in this method, the speed required for processing one sample requires about 25 minutes.
Speedup was required.

[0005] Also, in this method, when this method is performed in order to capture smaller cells from a sample containing a large solid such as squamous epithelial cells, the target cells collected on a filter are squashed. Epithelial cells and the like are collected so as to cover them, which makes them unsuitable for observing target cells.

In addition, it is necessary to keep the suction pressure constant. However, in the conventional ON-OFF control of the solenoid valve, a pressure pulsation is generated and the height of the suction bottle at the time of decompression suction through the suction bottle is set. (In general, the same height as the table on which the main body is installed, or the difference in height when the main body is installed on the table and the suction bottle is placed on the floor.) Was.

[0007] In addition, a filter for collecting target cells is attached, a sample is charged, and a solid content (including target cells) in the sample adheres to the inner wall of a funnel into which a washing / fixing solution is charged. There was a problem that the work of setting the filter and fixing the funnel was complicated.

[0008]

SUMMARY OF THE INVENTION In filtering and collecting cells from a suspension containing target cells, in particular, from a liquid biological sample, the cells are processed in a short time while maintaining the goodness of the obtained sample.

A method for selectively collecting target cells from a sample containing squamous epithelial cells and the like.

[0010] A method of adjusting the suction pressure which eliminates the pulsation of the suction pressure and is not affected by the installation height of the suction bottle.

A structure of a funnel that does not allow solids in a sample liquid to adhere to the inner wall of the funnel and a one-touch fixing method of a filter funnel.

[0012]

Means for Solving the Problems JP-A-2000-1467
The standardization disclosed in No. 82 (a good sample can be easily obtained by anyone without any experience) is examined and tested, and the cells are collected in a pure form and without denaturation, and are collected on a filter. To extract the minimum necessary elements for fixation and to create a sample with good reproducibility, the parts that require skill (work from the introduction of the washing solution after fixation of the sample, fixation solution to suction, and detection of the end point) The work was automatically processed, and simple tasks that did not require skill were manually performed. The work was clarified and separated.

[0013] When a cell suspension containing cells collected from various organs of a living body is filtered, the cells are collected on a filter, and a preparation for cytodiagnosis is prepared. In order to remove this, treatment was carried out using a mesh having a large pore size or a multilayer structure of a filter and a filter having a small pore size. The lower layer is provided with a filter for collecting target cells, and the upper layer is provided with a mesh or filter which collects large cell clumps (small tissue pieces) and non-cell components and transmits cells to be collected. It has a three-layer structure in which a strong material, for example, a stainless steel wire mesh filter is interposed between the two types of filters to facilitate filtration.

Similarly, in order to collect small cells such as lymphocytes and blood cells, the cells are treated with a combination of filters. In this case, a filter having a large pore size is used in the upper layer to collect target cells, and a filter having a smaller pore size than the upper layer is placed in the lower layer to collect small cells. It is a three-layer structure filter in which a strong material, for example, a stainless steel wire mesh filter is interposed between the two types of filters to facilitate filtration.

A branch pipe is provided in the suction pipe near the suction port of the funnel while continuously suctioning without performing ON-OFF control that causes pulsation. A pore is provided at the tip of the branch pipe,
A mechanism for adjusting the size of the hole is added. At the time of suction, constant air always enters through the fine holes, and as a result, the suction pressure of the funnel suction port becomes constant.

The funnel shape is a cylinder, and the inner diameter of the cylinder is slightly larger than the size (diameter) of the filter. A structure in which the lower cross section of the cylindrical funnel uniformly hits the frame of the filter. The cylindrical funnel has a structure with a flange below, and is equipped with a fixing jig that presses the flange from above from both sides.

[0017]

FIG. 1 is a work flow chart showing the sharing of automatic operation and manual operation on the apparatus side according to the present invention. First, the specimen is placed in a cell suspension such as physiological saline. For viscous samples, add a commercially available lysing agent to the cell suspension in advance. Prepare samples to be processed in this way. The apparatus turns on the power, and sets a filter (in some cases, a multilayer filter) and a funnel suitable for the sample.
The sample is dropped into the funnel together with the cell suspension, and the start switch is turned ON. The apparatus automatically starts suction and stops suction at the liquid level just above the filter. At this time, if the liquid level does not reach the above-mentioned level within a predetermined time (for example, 30 seconds), an alarm is issued (the alarm means that the sample is viscous and sufficient dissolving agent is not added). After reaching the liquid level and stopping the suction, immediately the cleaning liquid (injection liquid 1)
Is put into a fixed amount (variable) funnel and starts suction at the same time. The suction stops at the time when the liquid level is suctioned, and
Immediately, a fixed amount (variable) of a fixative (injection liquid 2 = generally using ethanol) is introduced, and after continuing the state for a fixed time (variable), suction starts and the liquid is sucked up to the above-mentioned liquid level An alarm is issued at that point to inform that the processing in the device has been completed. The operator removes the funnel, moves the target filter to a separate fixing solution tank, sets a new filter and funnel, inputs a new sample, and turns on the start switch. This series of operations is repeated, and when the last sample operation is completed, the timer switch attached to the apparatus is turned ON. This timer switch issues an alarm after the time (variable) for immersing the filter in the fixed tank. After this alarm, the filters in the fixing solution tank are collectively transferred to the next commercially available dye stabilizing solution tank, and the next timer switch attached to the apparatus is turned on. This timer issues an alarm after a certain period of time (variable) to indicate that all operations have been completed.

FIG. 2 is a conceptual diagram in the case of superposition filtration when fixing the funnel and the filter of the present invention, and FIG.
It is sectional drawing at the time of lap filtration when fixing the funnel of this invention and a filter. A multi-layer (three) filter is placed on the funnel and filter receiving portion, and a funnel with a cylindrical flange is placed on the filter. The flange of the funnel is fixed at one time from both sides by a funnel filter holder.

FIG. 3 is a conceptual diagram in the case of conventional filtration when fixing the funnel and the filter of the present invention.
FIG. 2 is a cross-sectional view of a conventional filtration when fixing the funnel and the filter of the present invention. Place the spacer in the funnel and filter receiver and place the filter. A funnel with a cylindrical collar is placed on it, and the flange of the funnel is fixed at once from both sides with a funnel filter holder.

FIG. 6 is a flowchart showing an adjustment mechanism for maintaining a constant suction pressure and a flow chart in a case where the suction pressure is attached to the suction line. A branch pipe is provided upward in the vicinity of a portion from the funnel filter section to the suction pipe through the electromagnetic valve, and a suction pressure adjusting section is provided at the tip thereof. The pressure adjusting unit has a hole that communicates with the outside air, and a threaded portion having a sharp end toward the hole is provided. The screw portion has a structure that can adjust the size of the hole. By tightening the screw to reduce the hole opening, or loosening the screw to increase the hole opening,
The suction pressure of the suction pump can be increased, not, or reduced.

[Brief description of the drawings]

FIG. 1 is a flowchart showing details of the apparatus of the present invention and manual operation.

FIG. 2 is a conceptual diagram in the case of the lap filtration method of the present invention.

FIG. 3 is a conceptual diagram in the case of a conventional filtration method of the present invention.

FIG. 4 is a view showing a state in which a funnel and a filter of the present invention are fixed.
It is sectional drawing of a lap filtration method.

FIG. 5 is a view showing a state in which the funnel and the filter of the present invention are fixed.
It is sectional drawing in the case of the conventional filtration.

FIG. 6 is a schematic diagram of a mechanism of a suction pressure adjusting unit of the present invention.

[Explanation of symbols]

 REFERENCE SIGNS LIST 1 funnel 2 funnel, filter holder 3 O-ring 4 filter with large pore size 5 spacer filter 6 filter with small pore size 7 funnel, filter receiving part 8 connection part to suction pipe 9 spacer 10 liquid level detection electrode 11 solenoid valve 12 suction pipe 13 Suction pressure adjustment unit 14 Suction bottle (waste liquid tank) 15 Suction pump 16 Connection part from suction pressure adjustment unit to suction pipe 17 External air intake hole adjustment screw 18 External air intake hole

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 1/30 G01N 33/48 P 33/48 G02B 21/34 G02B 21/34 G01N 1/28 U J ( 72) Inventor Masao Honma 141-17 Fuji, Shirai-machi, Inba-gun, Chiba Prefecture (72) Inventor Zenshiro Koyama 1162-3, Miyanogi-cho, Inage-ku, Chiba-shi AA27 BB11 CC01 FA01 FA11 4B063 QA19 QQ08 QR52 QR53 QS10 QS12 QS20 QS39

Claims (4)

[Claims] 1. A specimen from a biological sample is used as a cell suspension,
It is a device that semi-automatically performs a series of operations for collecting cells by filtering it and preparing a preparation for cytodiagnosis, and separates the parts to be automatically processed from the tasks to be performed manually. Specifically, first, the specimen must be manually placed in a cell suspension such as physiological saline. For viscous samples, add a commercially available lysing agent to the cell suspension in advance.
Put the specimen into the device together with the cell suspension. After the automatic processing of the device, remove the filter and put it into the cell sample fixation solution tank. After a certain period of time, transfer the filter from the cell sample fixing solution tank to the staining stabilizer solution tank. The device aspirates the cell suspension containing the injected sample, stops the aspiration when the liquid level reaches a liquid level sensor point (hereinafter referred to as point A) attached near the filter surface, and immediately removes the washing liquid. Inject and start liquid aspiration. When the liquid level reaches point A, the suction is stopped and the sample fixative is immediately injected. Stop for a certain period after injection, then start suction. An alarm is issued when point A is reached. A cell specimen fixing method in which this series of operations are all performed automatically. 2. A method for filtering a cell suspension containing cells collected from various organs of a living body, collecting the cells on a filter, and preparing a preparation for cytodiagnosis. A method of treating cells with a multilayer structure of a mesh (or filter) and a filter to remove cell components. Use a large pore size filter (or mesh) for the upper layer,
Place a small pore size filter in the lower layer. A method of performing suction filtration with a three-layer filter in which a strong material, for example, a stainless steel wire mesh filter is interposed between the two types of filters. 3. A branch pipe is provided in a suction pipe near the lower part of the funnel, near the funnel and after receiving the filter. A suction pressure adjusting device that has a fine hole at the tip of a branch pipe and adds a mechanism that can adjust the size of the fine hole. 4. A suction filter can be easily set,
Cylindrical funnel with a fixing flange that has a structure in which all collected cells etc. flow into the filter