Method for quantitatively analyzing fragmented red blood cells

A method for quantitatively analyzing fragmented red blood cells, comprising the steps of:

• adding a nucleic acid staining fluorochrome dye to a blood sample for preparation of a specimen containing particles;
• allowing the specimen to flow through a flow cytometer, and measuring a scattered light intensity and a fluorescent light intensity for each particle in the specimen;
• preparing a two-dimensional particle distribution diagram on the basis of the measured scattered light intensity and fluorescent light intensity;
• establishing a red blood cell distribution area on the two-dimensional distribution diagram, and counting particles falling within the red blood cell distribution area for determination of the total number A of red blood cells;
• establishing a fragmented red blood cell distribution area in the red blood cell distribution area, and counting particles falling within the fragmented red blood cell distribution area for determination of the number B of fragmented red blood cells;
• calculating a value B/A, and calculating a content by percentage F of the fragmented red blood cells from a conversion function F=f(B/A);
• establishing a microerythrocyte distribution area in a region in which the scattered light intensity is higher than in the fragmented red blood cell distribution area within the red blood cell distribution area;
• counting particles falling within this area for determination of the number C of microerythrocytes;
• and
• correcting the content by percentage F on the basis of CIA.

A method for quantitatively analyzing fragmented red blood cells, comprising the steps of:

• adding a nucleic acid staining fluorochrome dye to a blood sample for preparation of a specimen containing particles;
• allowing the specimen to flow through a flow cytometer, and measuring a scattered light intensity and a fluorescent light intensity for each particle in the specimen;
• preparing a two-dimensional particle distribution diagram on the basis of the measured scattered light intensity and fluorescent light intensity;
• establishing a red blood cell distribution area on the two-dimensional distribution diagram, and counting particles falling within the red blood cell distribution area for determination of the total number A of red blood cells;
• establishing a fragmented red blood cell distribution area in the red blood cell distribution area, and counting particles falling within the fragmented red blood cell distribution area for determination of the number B of fragmented red blood cells;
• and
• calculating a value B/A, and calculating a content by percentage F of the fragmented red blood cells from a conversion function F=f(B/A), wherein the conversion function F=f(B/A) is expressed as a linear equation F=m(B/A)+n wherein “m” and “n” are constants.

An apparatus for analyzing fragmented red blood cells, comprising:

• a flow cell for flowing a specimen containing particles therein, the specimen being prepared by adding a nucleic acid staining fluorochrome dye to a blood sample;
• a light source for emitting a light beam to the particles in the specimen;
• a first detector for detecting an intensity of scattered light generated from each particle;
• a second detector for detecting an intensity of fluorescent light generated from each particle;
• and
• a data processor for processing the detected intensities, wherein the data processor carries out,
• a first step of preparing a two-dimensional particle distribution diagram on the basis of the measured scattered light intensity and fluorescent light intensity;
• a second step of establishing a red blood cell distribution area on the two-dimensional diagram, and counting particles falling within the red blood cell distribution area for determination of the total number A of red blood cells;
• a third step of establishing a fragmented red blood cell distribution area in the red blood cell distribution area, and counting particles falling within the fragmented red blood cell distribution area for determination of the number B of fragmented red blood cells;
• and
• a fourth step of calculating a value B/A, and calculating a content by percentage F of the fragmented red blood cells from a conversion function F=f(B/A).