Method for preparing reticulocyte simulating particles and platelet simulating particles, and reference control

A method for preparing reticulocyte simulating particles, comprising:

• treating mammalian anucleated red blood cells, each of which has a volume of 60-120 fL, with a protein fluorescent dye having a carboxyl group activated by N-hydroxysuccinimide, the carboxyl group activated by N-hydroxysuccinimide of the protein fluorescent dye is covalently cross-linked with an activatable amino group on membrane surfaces of the mammalian anucleated red blood cells;
• and
• treating the mammalian anucleated red blood cells with a fixative to prepare the reticulocyte simulating particles, wherein the fixative is at least one selected from a group consisting of dichromate, formaldehyde, and glutaraldehyde,
• wherein the treating with the fixative is performed before or after the treating with the protein fluorescent dye;
• and
• obtaining scatter diagram of the prepared reticulocyte stimulating particles, wherein in a scatter diagram of the prepared reticulocyte simulating particles, the reticulocyte simulating particles have a same distribution as fresh blood reticulocytes in a scatter diagram of fresh blood, wherein the scatter diagram of the reticulocyte simulating particles is obtained by detecting the reticulocyte simulating particles in a RET channel of a blood analyzer;
• the scatter diagram of the fresh blood is obtained by detecting a fresh blood sample in the RET channel of the blood analyzer;
• and in both of the scatter diagrams, an abscissa represents fluorescence intensity and an ordinate represents forward scattered light intensity;
• wherein the protein fluorescent dye comprises at least one fluorescent dye selected from the group consisting of fluorescent dyes A, B, C, D, E, F, G, and H,
• structural formula of the fluorescent dye A is
• structural formula of the fluorescent dye B is
• structural formula of the fluorescent dye C is
• structural formula of the fluorescent dye D is
• structural formula of the fluorescent dye E is
• structural formula of the fluorescent dye F is
• structural formula of the fluorescent dye G is
• structural formula of the fluorescent dye H is