KR20040010136A - Transparent ruler tape for cell count - Google Patents

Abstract

PURPOSE: A transparent ruler tape for cell counting is provided, thereby accurately counting the number of low concentration cells, cheaply and easily manufacturing the transparent ruler tape and observing the ruler and cells simultaneously. CONSTITUTION: A transparent ruler tape for cell counting is prepared by drawing vertical and horizontal lines having the same distance such as 2,5, 1, 0.5 and 0.25 mm with ink or blade on the adhesive surface of a general adhesive transparent tape. The transparent ruler tape is adhered to a plate and a flask for tissue cultivation and the ruler and cells are simultaneously observed by an optical microscope.

Description

Transparent ruler tape for cell count}

Cell counts of red blood cells, white blood cells (neutrophils, eosinophils, lymphocytes), macrophages, epithelial cells, tumor cells, etc. The clinical significance of the measurement is significant in the diagnosis, post-treatment observation and prognosis of the disease. For example, the number of erythrocytes can be used to examine the presence or the degree of anemia, and since leukocytes appear to increase or decrease in many diseases, the coefficient is important for screening diseases, and especially the measurement of leukocyte count in cerebrospinal fluid is tuberculous meningitis. It is used as a differential diagnosis of bacterial meningitis and viral meningitis, and is a particularly important test item for diseases requiring urgent judgment and treatment. Cell counts have also been used in tissue slides, particularly in bone marrow tissue, with idiopathic thrombocythemia, reactive thrombocytosis, and idiopathic thrombocytopenic purpura, depending on the increase or decrease in the number of megakaryocytes per unit area in bone marrow tissue. Is important in the diagnosis. In addition, after culturing specific cells or strains for research purposes, it is necessary to measure the number of cells in order to success and secondary use of the culture.

At present, the counting method of cell number is largely divided into a trial method and an automated method. The trial method is a method of preliminary examination on a manual hemocytometer and counts blood cells without treatment, and stains and counts nuclei of blood cell cells with a staining agent. The principle of the manual hemocytometer is made of glass material and the sample is spread out by capillary phenomenon and consists of a certain volume space so that a certain amount can be entered and observed directly under an optical microscope to count the cell number. The above-mentioned automated method is a method of diluting a certain amount of blood and passing it through an ultrafine flow path to detect blood cells by electric resistance or scattered light and counting the number of blood cells, and is usually performed by an automated blood counting device. These counting methods differ in whether they are based on human vision or by machine, but are actually methods of calculating cell numbers. However, these counting methods have the following problems.

In the trial method, a coefficient error occurs according to cell misunderstanding due to hemolysis of red blood cells, aggregation of blood cells, and cell transformation, and skill of a measurer. In addition, in the trial method, it is difficult to discriminate the shape of the cells, and in many cases, it is necessary to confirm the number of cells by checking them with a smear stained sample. In particular, when measuring the number of cells in the urine or body fluid using a manual hemocytometer, the cells are deformed with time and the measured cell number decreases drastically. Can be. In addition, the existing manual hemocytometer is made of glass material to maintain the correct volume is well broken, there is a relatively expensive disadvantage.

The automated method is used for the measurement of blood cell counts, and due to the appearance of erythrocytes, coagulation of blood, platelet aggregation, poor hemolysis of erythrocytes, and the like, correct counts may not be obtained. If present, the measurement is inaccurate and measured by the trial method. Such a device is expensive and large.

[Reference: Transfusion Science 1997; 18 (4): 505-15, Transfusion1994; 34 (1): 35-8, Clinical Laboratory Haematology 2001; 23: 43-51, Transfusion 2000; 40: 513-20, Transfusion 1993 ; 33 (5): 409-12.]

The purpose of the present invention is to provide a compact, convenient, accurate, simple and low cost tool for the calculation of cell numbers of white blood cells, red blood cells, platelets, megakaryocytes, and tissue cells on clinical specimens, research specimens and tissue slides at low cost. In particular, it is possible to provide a tool for accurately measuring low concentration cell numbers.

In order to achieve the above object, the present invention is a single compartment made of a square, by the ink and the blade at the interval of 2.5 mm, 1 mm, 0.5 mm, 0.25 mm horizontally and vertically on a general transparent tape having an adhesive surface, respectively The scales entered the field of view of the microscope magnified at 40 times, 100 times, 200 times and 400 times respectively. In the case of the conventional manual hemocytometer, the accurate measurement requires a high degree of accuracy in order to make the space of the accurate volume, whereas in the case of the transparent lattice tape, the sample is dipped with an accurate pipette that has already been controlled. In order to make the measurement, it is easy to manufacture because the distance of one lattice is made to enter the field of view of the microscope. In addition, by attaching the tape to tissue slides, culture flasks and plates, it is possible to measure various types of cells based on the grid scale under a microscope.

1 is a plan view of a slide with a transparent lattice tape for cell number measurement according to the present invention and a plan view at the time of microscope observation. (A): Plan view of slide with transparent lattice tape, (B): 40 times magnification microscope field of view with 40 times transparent lattice tape, (C): 100 times magnification microscope field with 100 times transparent lattice tape, (D): 200 times magnification microscope field of view when attaching the transparent grid tape for 200 times, (e): 400 times magnification microscope field when attaching the transparent grid tape for 400 times

Figure 2 is an illustration of a cell culture plate and flask attached with a transparent grid tape for measuring cell number according to the present invention

<Description of the symbols for the main parts of the drawings>

Transparent lattice tape for cell number measurement 2. Normal slide

3, 6. Transparent Grid Tape for Cell Number Measurement

4. Plate for cell culture 5. Well

7. Flasks for Cell Culture

The present invention is a 40 times use (FIG. 1, B), 100 times (depending on the magnification of the microscope for observing the cells to be measured with a transparent grid tape for measuring the number of cells that can be adhered to the microscope slide, culture plate and culture flask) 1, c), 200 times (Fig. 1, D), 400 times (Fig. 1, E) are divided into. In the case of 40 times use, the diameter of one field of view is 5 mm when viewed under a 40 times magnification microscope, so that the distance between each horizontal and vertical lattice is 2.5 mm so that one section of 6.25 mm square can be observed in one field of view. By attaching ink or blades to the adhesive side of the tape, each grating and the cells to be observed were simultaneously observed in the microscope field when attached on the slide. Thus, the space | interval between each lattice was set to 1 mm for 100 times, 0.5 mm for 200 times, and 0.25 mm for 400 times. The thickness of the line of demarcation is irrelevant because the cells and the lattice are well observed in each microscope field of view, but the spacing of the grids is 2.5 mm for 40 times, 1 mm for 100 times, 0.5 mm for 400 times, and 0.5 mm for 400 times. Since it is 0.25 mm, it is preferable to produce 0.625 mm, 0.25 mm, 0.125 mm, and 0.0625 mm or less which are 25% of the space | interval between gratings, respectively.

The method of using the transparent lattice tape for measuring cell number is to drop a certain amount of sample on the slide, and then to dry it, and to stain the cells for easy observation under an optical microscope (Wright Giemsa stain or Hematoxylin Eosin stain, etc.) After attaching the transparent lattice tape for measuring the cell number, the cell number of the sample deposited based on the lattice of the tape can be counted by counting under the optical microscope. By dripping, the concentration can be calculated from the counted cell number and the cell number can be measured. When calculating the number of cells in the culture plate or flask, a transparent lattice tape for measuring the cell number was attached to the lower part of the culture plate (Fig. 2, a) or flask (Fig. 2, b), and then observed under an optical microscope to measure cells based on the grid. A method of counting and measuring numbers. When calculating the number of cells such as megakaryocytes on the tissue slide, a transparent lattice tape for measuring the cell number was attached on the tissue slide (Fig. 1, a), and the cell number per unit area was counted based on the area of the lattice. As such, the transparent lattice tape for cell number measurement can be conveniently and easily attached to slides, plates and flasks when cell number measurement is required in clinical and research samples.

Hereinafter, embodiments of the present invention are as follows.

Three normal blood samples were collected in a test tube containing anticoagulant K ₃ EDTA, and leukocyte counts were measured using an automatic hemocytometer, CellDyn 4000 (Abott, USA). Automatic blood counting device, Naegeotte chamber, manual blood counting device, and cell number measurement for this slide were measured 4 times each using a transparent grid tape, and the measured average white blood cell counts were compared. The cell grid for slide counting was used for 200 times (Fig. 1, D), and each sample was dipped on the slide by 1 μl or 10 μl using a micropipette, and then dried and stained by Wright Giemsa. A tape was attached and counted under a 200 times magnification optical microscope to calculate the cell number. Each measured white blood cell count is shown in Table 1, and the result of calculating the error [{(expected value-measured value) / expected value} × 100] for each measured white blood cell number (Table 2). Leukocyte measurement method according to the design was the most accurate. The regression equation for each white blood cell count was calculated from the expected value, and the correlation coefficient (R ² ) with the expected value was found.Therefore, there was no significant difference between 0.99901 using an automatic hemocytometer, 0.9984 using a manual hemocytometer and this transparent lattice tape. .

(Unit: / μl) Sample 1, white blood cell count 4523 / μl Sample 2, white blood cell count 6505 / μl Sample 3, white blood cell count 7540 / μl Expected value (/ μl) Automatic blood cell calculator Manual blood cell calculator Transparent grid tape Expected value (/ μl) Automatic blood cell calculator Manual blood cell calculator Transparent grid tape Expected value (/ μl) Automatic blood cell calculator Manual blood cell calculator Transparent grid tape 2262 2238 2218 2239 3253 3227 2938 3012 3770 3738 3562 3549 1131 1143 996 1108 1626 1654 1578 1592 1885 1868 1772 1758 565 601 537 552 813 858 756 794 943 998 907 921 283 314 254 262 407 439 389 382 471 506 427 441 141 183 126 129 203 266 184 192 236 281 209 220 71 117 62 67 102 148 92 89 118 176 34 105 35 0 28 30 51 0 43 50 59 94 51 54 18 0 14 15 25 0 19 22 29 0 24 25

(unit: %) Sample 1, white blood cell count 4523 / μl Sample 2, white blood cell count 6505 / μl Sample 3, white blood cell count 7540 / μl Expected value (/ μl) Automatic blood cell calculator Manual blood cell calculator Transparent grid tape Expected value (/ μl) Automatic blood cell calculator Manual blood cell calculator Transparent grid tape Expected value (/ μl) Automatic blood cell calculator Manual blood cell calculator Transparent grid tape 2262 1.06 1.95 1.02 3253 0.80 9.68 7.41 3770 0.85 5.52 5.86 1131 1.06 11.94 2.03 1626 1.72 2.95 2.09 1885 0.90 5.99 6.74 565 6.37 4.96 2.30 813 5.54 7.01 2.34 943 5.83 3.82 2.33 283 10.95 10.25 7.42 407 7.86 4.42 6.14 471 7.43 9.34 6.37 141 29.78 10.64 8.51 203 31.03 9.36 5.42 236 19.07 11.44 6.78 71 64.79 12.68 5.63 102 45.10 9.80 12.74 118 49.15 71.19 11.02 35 100.00 20.00 14.29 51 100.00 15.69 1.96 59 59.32 13.56 8.47 18 100.00 22.22 16.67 25 100.00 24.00 12.00 29 100.00 17.24 13.79

As described above, the present invention attaches a transparent adhesive tape, which is equally spaced, to a slide containing clinical and research specimens, a flask for bacterial culture, and a plate, and various cells such as white blood cells, red blood cells, platelets, megakaryocytes, and tissue cells in the optical microscope field. It can be used for the calculation of the number of cells, and it is possible to measure the exact low concentration cell number. The tape is easy to manufacture, low cost, and easy to use, making it easy to use for measuring various cell numbers in small laboratories and laboratories.

Claims (1)

The adhesive surface of the general transparent tape with adhesive strength is horizontally and vertically delineated with ink or blades at equal intervals (2.5, 1, 0.5, 0.25 mm), respectively, to adhere to slides, tissue culture plates and flasks, and then to an optical microscope. At the time of observation, the lattice and the cells can be observed at the same time, so that cell number measurement is possible.