SU1144031A1 - Erythrocyte sphericity index determination method - Google Patents

Abstract

A METHOD FOR DETERMINING THE ERYRTHROCHGG SPHERICAL INDICATOR based on dilution of whole blood, illuminating the suspension with a collimated light beam, recording the light fluxes of radiation that passed through the suspension, replicating so that, in order to simplify the method and ensure accuracy, they are recorded in a solid angle O; Q: 2.4 10 cf minimum value of the luminous flux that passed through the suspension in the spectral range of 450-800 nm before and after the substance that forms the red blood cells is introduced into the sphere without changing their volume, and then in relation to the measured minimum (L values of the light fluxes court r on the desired value.

Description

 N

The invention relates to blood spectrophotometry and can be used to diagnose in hematology. A known method for determining the sphericity parameter of aqueous suspensions, based on the illumination of a suspension by a collimated beam of light, measures the mm luminance scattered by particles from two angles of 140 and 90 ° D. The disadvantage of this method is its applicability only to suspensions part of the index of premature lies in the range of P 1.1-1.15, which makes it unacceptable for erythrocytes, in which P is 1.07. There is a known method for determining the sphericity index of particles p, the basis of which is measuring the logarithmic derivative of luminance angle, in the interval of angles 135-140 2. The disadvantage of this method is its complexity, which consists in the need to move the scattered radiation receiver, in the corners 135- 140 with high angular resolution to determine the logarithmic derivative of luminance. With insufficient angular resolution increases the measurement error. The need to measure small light fluxes as compared with the incident light and {exceeding the background no more than 10 times in the angles of 135–140tac. E increases the measurement error. The disadvantages associated with complexity and inaccuracies make the method unacceptable for widespread use in hematology. The closest in technical essence to the present invention is a method for determining the red blood cell sphericity, based on the dilution of whole blood, placing the erythrocyte suspension in two cuvettes, creating a laminar flux in one of the cuvettes, illuminated by their radiation with a wavelength of 415 nm, where erythrocytes have an absorption maximum, are determined by the absorbance values of the optical densities of two cells in one of which red blood cells are in a state of rest, and in the other direction they move in a flow, and determine the average sphericity index of red blood cells in Gz. The disadvantage of this method is its complexity, due to the need to ensure the orientation of red blood cells in one of the cuvette, as well as an error of about 5% in determining the optical density due to photochemical reactions when exposed to light from 415 nm due to the formation of hemoglobin derivatives. In addition, when creating a laminar flow, the erythrocytes are deformed and destroyed. The purpose of the invention is to simplify the method and increase the accuracy of measurements. The goal is achieved according to the method of determining the sphericity index of erythrocytes, based on the dilution of whole blood, illuminating the suspension with a collimated light beam, recording the light fluxes of radiation transmitted through the suspension, recorded in a solid angle of 2.4 the minimum value of the light flux that led through the suspension in the spectral the interval 450-800 nm before and after the introduction into suspension of a substance that forms red blood cells into the sphere without changing their volume, and then in relation to the measured minimum The values of luminous fluxes is judged on the desired value. When registering the minimum transmitted light flux through a suspension of erythrocytes, the determined physical quantity is the attenuation index: where B is the thickness of the suspension layer; incident luminous flux; transmitted minimum light flux; in one of the wavelengths in the spectral range of 450-800 nm. In this case, ("Ro. I Fog, g-en-; HS2 g6n-, where Ct is the indicator of the attenuation of the luminous flux Fd | when it passes through a layer of thickness B of the erythrocyte suspension; the indicator of the attenuation of the light flux F during its passage through the layer the thickness of the suspension of erythrocytes with the addition of a substance that forms the erythrocytes into the sphere, 1 is the minimum value of the luminous flux passed through the suspension of erythrocytes, Tj is the maximum value of the luminous flux passed through the suspension of erythrocytes with the addition of the substance forming the erythrocytes into the sphere. It goes, that. It, / f,. 2 bp og / sr, C use of 1aduyroochny schedule of dependence -t .- (p), obtained by the control method, determine the indicator of sphericity of erythrocytes R. Selection of the spectral interval 450-800 nm is due to the fact that the possible shnimalnye values of the luminous flux, and P3 passed through the suspension of viewers with different values of the sphericity index p before and after the introduction of the red blood cell forming substance into it, in an angle of 0 b $ 2.4, are in spectral region of 450-800 nm, and therefore, due to the weak absorption bands, the function The mutual mutual transformations of erythrocyte hemoglobin are insignificant and do not introduce errors in determining p. The reception angle, 4 10 sr, provides a measure of the attenuation index & transmitted radiation in the specified spectral interval with a minimum error (1-2%), arising mainly due to the contribution of scattered forward radiation, which determines the accuracy of measuring the sphericity index p of erythrocytes 1-2%. . B as a substance that forms red blood cells into the sphere without changing their volume, use lecithin, as a diluent, mTa3Nry or a physiological solution, which guarantees the invariance of the form of red blood cells at 18-22 ° C. Figure 1 shows the calibration curve; Fig. 2 shows a graph of the spectral dependence of the light flux ratio for erythrocytes with various sphericity indices. The calibration graph (figure 1 determines the erythrocyte sphericity index p by the ratio 1 1.4 attenuation index 1 for the corresponding minimum values of the light fluxes Pdim the erythrocyte passed through the suspension () and the lecithin added suspension (Pj), while plotting the sphericity index value p, equal to the ratio of the maximum size of the erythrocyte to the minimum, is determined by a microscopic method. The average diameter of the disk-shaped erythrocytes, rf, is then determined by spherical Jc (p and based on the equality of their volumes, the average value of the erythrocyte sphericity is determined Figure 2 shows the coincidence of Pf / 9d and g Lp red blood cells (points) with the corresponding p values and oblate rotation ellipsoids (solid lines). Example: The erythrocyte sphericity index, Three different human blood samples were used. Measurements were carried out at 18-22 C. Whole blood was diluted 2000 times with physiological saline, placed in a flat cuvette about 1 cm thick. The suspension was illuminated with a collimated light beam (angle 2.440 sr source, monochromator dispersion 50 A / mm). The luminous flux of transmitted radiation in a solid angle of 2.4 "10 sr is measured in the spectral range 450-800 nm, the minimum value of the transmitted luminous flux 1 is recorded. Then, licitin in the amount of 30 mg / l is added to the cuvette, and the measurements are repeated, registering the minimum value of the transmitted light flux -P. The indicator shows the sphericity of red blood cells in the table. The table shows the experimental results of the determination for pex blood samples of a person. Measurements to improve the accuracy of 2-5 important diagnos nical parameters of blood.

0.66 0.8A6 0.96

1.2 1.65 1.38

e, / Er

Continued Tabl Sample

Indicators

J.IZII

five

p2,6 53,7

p (control) 2.5 4.6 3.5

ten

 Vg

R

YU

1

Ni to

Claims (1)

THE METHOD FOR DETERMINING THE INDICATOR OF THE REDUCTION SPACE OF ERYTHROCYTES, based on the dilution of whole blood, illumination of the suspension with a collimated beam of light, registration of light fluxes of radiation transmitted through the suspension, characterized in that, in order to simplify the method and increase accuracy, they are recorded in a solid angle of 0 <Q ί 2.4 "10 ~ ^ cf the minimum value of the light flux passing through the suspension in the spectral range 450-800 nm before and after the introduction of the substance forming red blood cells into the sphere without changing their volume, and then by the ratio The measured minimum luminous flux values are used to judge the desired value. SLL ..., 1144031 ί