JPH04104036A - Manufacture of bone marrow specimen - Google Patents

Abstract

PURPOSE:To reduce the overlapping of cells and to make dispersion uniform by dispersing the cells in bone marrow liquid into diluent liquid, collecting the specified number of the cells, and performing centrifugal coating. CONSTITUTION:Bone marrow liquid is sucked from an organism with a syringe 3 containing fetal bovine serum/physiological saline wherein diluent anticoagulent is added. Diluent liquid 2 of the bone marrow is transferred into a test tube 4, pipetting is performed by using a pipet 5 and cells are made to float in the liquid 2. When the state wherein the cells are uniformly floating in the diluent liquid 2 of the bone marrow is obtained, the pipetting is finished. Then, the liquid 2 undergoes centrifugal separation, and supernatant 2a is removed. The cells having the amount required for centrifugal coating are collected. The diluent liquid 2b from which the supernatant 2a is removed is dropped on slide glass and spread into a uniform layer by the centrifugal coating. Then, Romanowsky stain is performed. In this way, the specimen wherein the overlapping of the cells is less and the dispersion is uniform can be prepared.

Description

[Detailed Description of the Invention] (a) Industrial Application Field This invention relates to a method for producing a bone marrow specimen, and more specifically,
The present invention relates to a method of producing a bone marrow specimen that is passed for classification by an automatic classification device.

(B) Conventional technology The conventional method for producing bone marrow specimens involves collecting bone marrow fluid from an animal, placing the collected bone marrow fluid on a slide glass, pressing it lightly with the front edge of the sliding glass to slowly smear it, and then removing the cells. is being stained.

(c) Problems to be Solved by the Invention Since bone marrow fluid has a high viscosity, in the conventional bone marrow specimen manufacturing method described above, cells in the specimen often overlap and are unevenly distributed. The overlapping and non-uniform distribution of cells differs depending on the skill of the specimen creator, and even the same specimen creator changes from time to time, and is not always the same.Especially, when using an automatic classification device, When automatically classifying cells in a specimen, specimens with overlapping cells or uneven distribution are inappropriate, and there is a problem in that accurate classification cannot be performed.

This invention was made in view of the above, and is aimed at providing a method for producing bone marrow specimens that allows a specimen with uniform distribution (uniform monolayer specimen) with few overlapping cells to be obtained.

(d) Means for solving the problem
It consists of the steps described in section -v.

i: a first step of aspirating bone marrow fluid from a living body with a syringe containing a diluent; ii: a second step of pipetting the bone marrow diluent diluted with the diluent; if: this pipetting a third step of centrifuging the diluted bone marrow solution and removing the supernatant to collect a predetermined amount of cells; tv: a fourth step of centrifugally smearing the collected cells;
2. A fifth step of Wright staining the centrifuged smeared cells.

In the method for producing a bone marrow specimen of the present invention, cells in bone marrow fluid are once dispersed in a diluent by pipetting, and a predetermined number of cells required for centrifugal smearing is collected by centrifugation. Since this is spread into a uniform monolayer by centrifugal smearing, there is less overlapping of cells and uniform dispersion can be achieved. In addition, there is no difference depending on the skill of the creator, and the sameness can be maintained at all times.

(e) Examples An embodiment of the present invention will be described below based on the drawings.

FIG. 1 is a diagram illustrating aspiration of bone marrow fluid.

1 is a small bone marrow puncture needle, which punctures the third to fifth sternum of the dog. Bone marrow fluid was diluted with EDTA as a diluent anticoagulant.
Aspirate with syringe 3 containing approximately 1 d of 30% tallow serum/saline 2° plus -2K.

Bone marrow diluent 2 diluted with tallow calf serum/physiological saline is
Transferred to test tube 4 [see Figure 2(a)].

Next, the bone marrow diluent 2 is pipetted using the pipette 5, and the cells are suspended in the bone marrow diluent 2 [see FIG. 2(b)].

When the cells become uniformly suspended in bone marrow diluent 2 [
Refer to FIG. 2(C)], and end the pipetting. Then, this bone marrow diluted solution 2 is centrifuged at 800 rpm for 5 minutes, and the supernatant 2a is removed so that the cell components account for about 70%, thereby collecting the amount of cells necessary for centrifugal smearing. (see figure).

The bone marrow diluted solution 2b from which the supernatant 2a has been removed is dropped onto a slide glass 6 [see FIG. 3(a)] and spread into a uniform monolayer by centrifugal smearing [see FIG. 3(b)]. The specimen 7 thus obtained is subjected to Romanovsky staining, and in this example, Wright staining.

FIG. 4 shows an enlarged image of the specimen 7 thus obtained. , it can be confirmed that there is little overlapping of cells and the distribution is uniform, and in particular, overlapping of nucleated cells (marked with arrows) is avoided. Therefore, automatic classification by an automatic classification device can also perform accurate classification. of course,
The specimen is also suitable for visual classification under a microscope.

(f) As described in detail of the invention, the method for producing bone marrow specimens of this invention includes:
The first step is to aspirate bone marrow fluid from a living body using a syringe containing a diluent.
Step 1, a second step of pipetting the diluted bone marrow solution diluted with the diluent, and a step of centrifuging the pipetted diluted bone marrow solution and collecting a predetermined amount of cells from which the supernatant has been removed. This process consists of step 3, a fourth step of centrifugally smearing the collected cells, and a fifth step of Wright staining the centrifugally smeared cells/cells, so there is little overlap of cells and they are dispersed. has the advantage of being able to produce uniform specimens. It also has the advantage that identity can always be ensured regardless of the skill of the specimen preparer.

[Brief explanation of the drawing]

Figure 1 is a diagram illustrating the aspiration of bone marrow fluid in an embodiment of the present invention, Figure 2 (a), Figure 2 (b), Figure 2 (
C) and Figure 2 are diagrams explaining the treatment of bone marrow dilution in the same example, respectively, and Figures 3 (a) and 3 (b) are diagrams showing the centrifugal smearing of cells in the same example, respectively. FIG. 4 is a diagram schematically showing an enlarged image of the specimen obtained in the same example. 2: Bone marrow dilution solution, 2゛: dilution solution, 3: syringe,
5: Pipette. Patent Applicant: OMRON Co., Ltd. Agent, Patent Attorney Shigeru Nakamura Nobuo Nakamura Diagram No. 3-5 Chest Diagram (a) Diagram (a) Diagram (b) 5: Vipe Diagram (C) Figure (d) Figure

Claims (1)

[Claims] (1) A first step of aspirating bone marrow fluid from a living body with a syringe containing a diluent, a second step of pipetting the bone marrow diluent diluted with the diluent, and a second step of pipetting the bone marrow diluent diluted with the diluent. A third step is to centrifuge the solution and remove the supernatant to collect a predetermined amount of cells, a fourth step is to centrifugally smear the collected cells, and a fifth step is to Wright-stain the centrifugally smeared cells. A bone marrow specimen manufacturing method comprising the following steps.