CN114839011A - Preparation method of reticulocyte quality control substance based on fluorescence principle - Google Patents

Abstract

The invention relates to the technical field of blood detection quality control substances, in particular to a preparation method of a reticulocyte quality control substance based on a fluorescence principle, which comprises the following steps: washing and separating the anticoagulated blood to obtain non-nucleated red blood cells; treating and fixing the anucleated red blood cells by using a cell curing agent to obtain cured red blood cells; carrying out curing agent removing treatment on the cured red blood cells to obtain red blood cell products; the red blood cell product is washed and then stored in a cell maintenance solution to obtain the reticulocyte simulant, and the prepared reticulocyte simulant is combined with red blood cells to prepare the reticulocyte quality control substance, so that the problem that the preparation cost is increased because protein fluorescent dye is needed to dye the cells when the reticulocyte is prepared is solved.

Description

A method for preparing reticulocyte quality control substance based on fluorescence principle

technical field

The invention relates to the technical field of blood detection quality control substances, in particular to a method for preparing a reticulocyte quality control substance based on a fluorescence principle.

Background technique

Reticulocytes are immature erythrocytes that are intermediate between late immature erythrocytes and mature erythrocytes. It is an important indicator to reflect the hematopoietic function of the erythroid of the bone marrow and to judge the curative effect of anemia and related diseases. Quality controls are used to check the performance of analytical instruments or methods and can ensure the reliability of laboratory results. The nature of the control samples should be as close as possible to the actual clinical samples tested, while at the same time ensuring suitability for the test system being monitored.

At present, the prior art (CN112986589A) discloses a method for preparing reticulocyte-simulating particles and platelet-simulating particles and a quality control material. The method for preparing reticulocyte mimetic particles of the present application uses N-hydroxysuccinimide-activated protein fluorescent dye to stain mammalian erythrocytes with a volume of 60-120 fL, and fix the mammalian erythrocytes to prepare reticulocyte mimetic particles. particle. The platelet mimetic particles were prepared using mammalian erythrocytes with a volume of 2-25fL, and the rest were the same as the reticulocyte mimetic particles. In the preparation method of the present application, N-hydroxysuccinimide-activated protein fluorescent dye is used to stain mammalian red blood cells of different volumes to obtain reticulocyte-simulating particles and platelet-simulating particles, respectively. The distribution of blood reticulocytes, reticulated platelets and platelet scatter plots; the prepared simulated particles have good stability and do not interfere with the counting and classification of other cell particles.

In the above manner, when preparing reticulocytes, it is necessary to use protein fluorescent dye to dye the cells, thereby increasing the manufacturing cost.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a method for preparing a reticulocyte quality control substance based on a fluorescence principle, which aims to solve the problem of increasing the production cost by using protein fluorescent dyes to dye cells when preparing reticulocytes.

To achieve the above purpose, the present invention provides a method for preparing a reticulocyte quality control substance based on a fluorescence principle, comprising the following steps:

Wash and separate anticoagulated blood to obtain anucleated red blood cells;

using a cell solidifying agent to process and fix the anucleated red blood cells to obtain solidified red blood cells;

The solidified red blood cells are treated with a de-solidifying agent to obtain a red blood cell product;

The erythrocyte product is washed and stored in a cell maintenance solution to obtain a reticulocyte simulant.

Wherein, the specific method of washing and separating anticoagulated blood to obtain anucleated red blood cells is:

Mix anticoagulant with blood to get anticoagulated blood;

The anticoagulated blood is washed and separated to obtain anucleated red blood cells.

Wherein, the specific method of de-solidifying the solidified erythrocytes to obtain the erythrocyte product is as follows:

After centrifuging the solidified red blood cells, remove the supernatant to obtain red blood cell pellets;

A cell maintenance solution and a de-solidifying agent solution are added to the red blood cell pellet, and the solution is allowed to stand for 10 minutes to 1 hour to obtain a red blood cell product.

Wherein, the anticoagulant includes EDTA anticoagulant or citric acid anticoagulant;

The cell fixative includes aldehyde cell fixative and base solution;

The cell solidifying agent also includes any one of a cell spheroidizing agent and a surfactant;

The cell curing agent also includes any one of a protein salting-out agent and a heavy metal salt, specifically including zinc sulfate, potassium sulfate, anhydrous sodium sulfate, ammonium sulfate, magnesium sulfate, zinc fluoride, barium chloride, and chlorine hexahydrate. Any one or more of cobalt;

The surfactants include cationic surfactants, anionic surfactants and nonionic surfactants.

Wherein, the cationic surfactant includes a quaternary ammonium salt surfactant, and specifically includes any one of cetyl ammonium bromide and dodecyl trimethyl ammonium chloride;

The anionic surfactants include sulfonate surfactants or sulfate ester salt surfactants, specifically including sodium dodecyl sulfate, sodium dodecyl sulfonate and sodium dodecyl benzene sulfonate. any one;

The nonionic surfactant includes polyoxyethylene or polyol fatty acid ester surfactant, specifically including any one of polyethylene glycol and dodecyl-β-D-maltoside;

The aldehyde cell fixative includes any one or more of formaldehyde, glutaraldehyde, o-phthalaldehyde and glyoxal;

The bottom solution includes any one of sodium chloride solution, borate buffer, and phosphate buffer.

Wherein, the dosage of the cell solidifying agent is 5-20 times the volume of the enucleated red blood cells.

Wherein, the osmotic pressure of the cell maintenance solution is 300±20mOsm/kgH0, the pH value is 7.3±0.1, and the cell maintenance solution includes antibiotics, polysaccharides, bovine serum albumin, sodium chloride, sodium citrate and citric acid;

The antibiotics include any one or more of penicillins, aminoglycosides and amide antibiotics;

The polysaccharide includes any one or more of sucrose, glucose, fructose, galactose, lactose, maltose, starch and dextrin.

The method for preparing a reticulocyte quality control substance based on the fluorescence principle of the present invention comprises washing and separating anticoagulated blood to obtain non-nucleated red blood cells; using a cell solidifying agent to process and fix the non-nucleated red blood cells to obtain solidified red blood cells; The solidified erythrocytes are treated with a de-solidifying agent to obtain a erythrocyte product; the erythrocyte product is washed and stored in a cell maintenance solution to obtain a reticulocyte simulant, and the prepared reticulocyte simulant is prepared by combining with erythrocytes. The reticulocyte quality control material solves the problem that the cells need to be dyed with protein fluorescent dyes when preparing reticulocytes, which increases the production cost.

Description of drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the following briefly introduces the accompanying drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only These are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can also be obtained according to these drawings without creative efforts.

FIG. 1 is a flow chart of a method for preparing a reticulocyte quality control substance based on a fluorescence principle provided by the present invention.

FIG. 2 is a test picture of the reticulocyte simulant prepared by the method of the present invention on a blood analyzer with the principle of nucleic acid fluorescence analysis.

3 is a test picture of the reticulocyte simulant prepared in the first embodiment on a blood analyzer with the principle of nucleic acid fluorescence analysis.

4 is a test picture of the reticulocyte quality control substance prepared by adding the erythrocyte simulant to the reticulocyte simulant prepared in the first embodiment on a blood analyzer with the principle of nucleic acid fluorescence analysis.

5 is a test picture of the reticulocyte quality control substance prepared by adding the erythrocyte simulant to the reticulocyte simulant prepared in the second embodiment on a blood analyzer with the principle of nucleic acid fluorescence analysis.

6 is a test picture of the reticulocyte quality control substance prepared by adding the erythrocyte simulant to the reticulocyte simulant prepared in the third embodiment on a blood analyzer with the principle of nucleic acid fluorescence analysis.

7 is a test picture of the reticulocyte quality control substance prepared by adding the erythrocyte simulant to the reticulocyte simulant prepared in the fourth embodiment on a blood analyzer with the principle of nucleic acid fluorescence analysis.

Detailed ways

The following describes in detail the embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein the same or similar reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary, and are intended to explain the present invention and should not be construed as limiting the present invention.

Please refer to FIG. 1 to FIG. 7 , the present invention provides a method for preparing a reticulocyte quality control substance based on a fluorescence principle, comprising the following steps:

S1 washes and separates anticoagulated blood to obtain anucleated red blood cells;

The specific method is as follows: S11 mixes the anticoagulant with blood to obtain anticoagulated blood;

Specifically, the anticoagulant includes an EDTA anticoagulant or a citric acid anticoagulant;

S12 washes and separates the anticoagulated blood to obtain anucleated red blood cells.

S2 uses a cell solidifying agent to process and fix the anucleated red blood cells to obtain solidified red blood cells;

Specifically, the dosage of the cell fixative includes 5-20 times the volume of the anucleated red blood cells, and the greater the dosage of the cell fixative, the faster the reaction process.

The cell fixative includes aldehyde cell fixative and base solution;

The cell solidifying agent also includes any one of a cell spheroidizing agent and a surfactant;

The cell curing agent also includes any one of a protein salting-out agent and a heavy metal salt, specifically including zinc sulfate, potassium sulfate, anhydrous sodium sulfate, ammonium sulfate, magnesium sulfate, zinc fluoride, barium chloride, and chlorine hexahydrate. The role of protein salting out agent/heavy metal salt is to change the protein structure on the cell, increase the viscosity, improve the ability to bind the dye, thereby enhancing the fluorescence intensity. The reticulocyte simulant can also be prepared without adding protein salting-out agent/heavy metal salt, but the fluorescence intensity of the cells will be slightly weaker, which is reflected in the instrument as the reticulocyte mass is closer to the erythrocyte mass. After adding protein salting out agent, the reticulocyte mass and the red blood cell mass can be separated more openly, which is helpful for the identification of the instrument. It is advisable to use heavy metal salts with a final concentration of 0.01%-1%. For example, ammonium sulfate at a final concentration of 0.1%-1% (w/v), cobalt chloride hexahydrate at a final concentration of 0.1%-1% (w/v), and a final concentration of 0.1%-1% ( w/v) barium chloride etc., also can be any combination of above-mentioned reagents;

The surfactants include cationic surfactants, anionic surfactants and nonionic surfactants.

Described cationic surfactant includes quaternary ammonium salt surfactant, specifically includes cetyl ammonium bromide, dodecyl trimethyl ammonium chloride, etc.;

The anionic surfactants include sulfonate surfactants or sulfate ester salt surfactants, specifically including sodium dodecyl sulfate, sodium dodecyl sulfonate, and sodium dodecyl benzene sulfonate, etc. ;

The nonionic surfactants include polyoxyethylene or polyol fatty acid ester surfactants, specifically including polyethylene glycol with a molecular weight of 1000-20000, dodecyl-β-D-maltoside, and the like;

The aldehyde cell fixative includes any one or more of formaldehyde, glutaraldehyde, o-phthalaldehyde and glyoxal; wherein, glutaraldehyde or a combination of glutaraldehyde and formaldehyde is preferred as the aldehyde Cell fixative; the concentration of the aldehyde cell fixative usually needs to be determined according to the time of action, and the final concentration can be 0.01%-2%. For example, formaldehyde at a final concentration of 0.01%-1% (v/v), glutaraldehyde at a final concentration of 0.001%-0.1% (v/v), and a final concentration of 0.01%-2% (v/v) can be used ) glyoxal, (w/v) o-phthalaldehyde with a final concentration of 0.01%-2%, etc., any combination of the above reagents can also be used; the action time of aldehyde reagents should not be too long, otherwise the red blood cells will be fixed. For the cells that are difficult to dissolve, red blood cells cannot be hemolyzed by hemolytic agents, and will be recognized as white blood cells when detected on the instrument, which affects the cell count; the action time of aldehyde reagents is usually 10 minutes to 2 hours, and the action time depends on the aldehyde reagents. depending on the concentration.

After adding the aldehyde cell fixative to the purified red blood cells, the cell shape and structure are changed under the action of cell spheroidizing agent or surfactant, and at the same time, the cells are fixed under the action of aldehydes and other fixatives. The change of cell shape is that the cell changes from a concave cake shape to a spherical shape, and the cell surface area increases; in this process, the cell membrane or protein molecules in the cell are fixed under the action of aldehydes, so that the protein has a certain binding fluorescent nucleic acid. The ability of dyes, even if cells have the ability to bind fluorescent nucleic acid dyes;

The final concentration of the surfactant in use is 0.005%-0.05% (w/v). For example, dodecyltrimethylammonium chloride at a final concentration of 0.005%-0.05% (w/v), sodium lauryl sulfate at a final concentration of 0.005%-0.05% (w/v), Dodecyl-β-D-maltoside at a final concentration of 0.005%-0.05% (w/v), or any combination of the above reagents;

The bottom solution is plasma isotonic solution, which can be sodium chloride solution or other buffer solution. For example, 0.9% (w/v) NaCl solution or borate buffer, phosphate buffer and the like can be used.

S3 performs de-solidifying agent treatment on the solidified red blood cells to obtain a red blood cell product;

Specifically, in S31, the solidified red blood cells are centrifuged and the supernatant is removed to obtain red blood cell pellets;

S32 Add a cell maintenance solution and a de-solidifying agent solution to the red blood cell pellet, and let it stand for 10 minutes to 1 hour to obtain a red blood cell product.

Specifically, the de-solidifying agent solution can be a reagent that can react with an aldehyde reagent, such as a protein reagent, and a bovine serum albumin solution with a concentration of 30%-50% (w/v) can be used.

In S4, the erythrocyte product is washed and stored in a cell maintenance solution to obtain a reticulocyte simulant.

Specifically, the osmotic pressure of the cell maintenance solution is 300±20mOsm/kgH20, the pH value is 7.3±0.1, and the cell maintenance solution includes 0.04%-0.2% (w/v) antibiotics, 0.5%-2% (w/v) /v) polysaccharides, 2%-5% (w/v) bovine serum albumin, 0.5%-1% (w/v) sodium chloride, 0.01%-2% (w/v) sodium citrate and 0.0001% -0.5% (w/v) citric acid;

Described antibiotics include any one or more of penicillin, aminoglycoside and amide antibiotics; specifically, one or more of penicillin, benzin penicillin, gentamicin, streptomycin and chloramphenicol can be used. combination of two or more. When the selection of antibiotics is the combination of the above two or more, the ratio between them can be any ratio;

The polysaccharide includes any one or more of sucrose, glucose, fructose, galactose, lactose, maltose, starch and dextrin, and when the selection of the polysaccharide is the above two or more combinations, the ratio between them can be Any ratio.

Example 1:

Fresh anticoagulated pig blood was collected using anticoagulant. Pig blood was centrifuged at 2000 rpm for 10 minutes. The plasma, platelet layer and leukocyte layer are removed after centrifugation, leaving purified red blood cells. Add 10 times the volume of erythrocyte volume to the erythrocytes, and place it at room temperature for 40 minutes after mixing, and mix it every 10 minutes. After the treatment, centrifuge at 2000 rpm for 5 minutes, pour off the supernatant, add cell maintenance solution 3 times the volume of the remaining cells, add 5% of the total volume of the de-solidifying agent solution after mixing, and act at room temperature for 30 minutes. Repeat washing the cells two to three times with the cell maintenance solution, and then transfer the cells to an appropriate amount of cell maintenance solution for preservation, which can be used as a reticulocyte simulant. Red blood cell mimics, white blood cell mimics, platelet mimics, and nucleated red blood cell mimics can be added to prepare the required quality controls. Add erythrocyte simulant to 4 × 10 ¹² cells/L, adjust the reticulocyte ratio to 5% to 6%, and use a blood analyzer based on nucleic acid fluorescence analysis principle for detection. Reticulocyte simulant) and Figure 4 (the left cell mass in the scatter diagram is an erythrocyte mimic, and the right cell mass is a reticulocyte mimic) only provide pictures of the RET channel.

Example 2:

Fresh anticoagulated pig blood was collected using anticoagulant. Pig blood was centrifuged at 2000 rpm for 10 minutes. The plasma, platelet layer and leukocyte layer are removed after centrifugation, leaving purified red blood cells. Add 10 times the volume of erythrocytes to the erythrocytes, and place the cell solidifying agent 2 at room temperature for 30 minutes after mixing, and mix once every 10 minutes. After the treatment, centrifuge at 2000 rpm for 5 minutes, pour off the supernatant, add cell maintenance solution 3 times the volume of the remaining cells, add 5% of the total volume of the de-solidifying agent solution after mixing, and act at room temperature for 30 minutes. Repeat washing the cells two to three times with the cell maintenance solution, and then transfer the cells to an appropriate amount of cell maintenance solution for preservation, which can be used as a reticulocyte simulant. Red blood cell mimics, white blood cell mimics, platelet mimics, and nucleated red blood cell mimics can be added to prepare the required quality controls. Add erythrocyte simulant to 4×10 ¹² cells/L, and adjust the ratio of reticulocytes to 5% to 6%. Use a blood analyzer based on the principle of nucleic acid fluorescence analysis for detection. The results are shown in Figure 5 (the cells on the left of the scatter diagram in the figure The cluster is an erythrocyte mimetic, and the cell cluster on the right is a reticulocyte mimetic) only the pictures of the RET channel are provided.

Example 3

Fresh anticoagulated pig blood was collected using anticoagulant. Pig blood was centrifuged at 2000 rpm for 10 minutes. The plasma, platelet layer and leukocyte layer are removed after centrifugation, leaving purified red blood cells. 15 times the volume of erythrocytes was added to the erythrocytes, and the cell solidifying agent 3 was placed at room temperature for 30 minutes after mixing, and the mixture was mixed every 10 minutes. After the treatment, centrifuge at 2000 rpm for 5 minutes, pour off the supernatant, add cell maintenance solution 3 times the volume of the remaining cells, add 5% of the total volume of the de-solidifying agent solution after mixing, and act at room temperature for 30 minutes. Repeat washing the cells two to three times with the cell maintenance solution, and then transfer the cells to an appropriate amount of cell maintenance solution for preservation, which can be used as a reticulocyte simulant. Red blood cell mimics, white blood cell mimics, platelet mimics, and nucleated red blood cell mimics can be added to prepare the required quality controls. Add erythrocyte simulant to 4×10 ¹² cells/L, adjust the reticulocyte ratio to 5% to 6%, and use a blood analyzer based on nucleic acid fluorescence analysis principle for detection. The cluster is an erythrocyte mimetic, and the cell cluster on the right is a reticulocyte mimetic) only the pictures of the RET channel are provided.

Example 4:

Fresh anticoagulated pig blood was collected using anticoagulant. Pig blood was centrifuged at 2000 rpm for 10 minutes. The plasma, platelet layer and leukocyte layer are removed after centrifugation, leaving purified red blood cells. The cell solidifying agent 4 was added to the red blood cells in an amount 10 times the volume of the red blood cells. After mixing, the cells were placed at room temperature for 1 hour, and the mixture was mixed every 10 minutes. After the treatment, centrifuge at 2000 rpm for 5 minutes, pour off the supernatant, add cell maintenance solution 3 times the volume of the remaining cells, add 5% of the total volume of the de-solidifying agent solution after mixing, and act at room temperature for 30 minutes. Repeat washing the cells two to three times with the cell maintenance solution, and then transfer the cells to an appropriate amount of cell maintenance solution for preservation, which can be used as a reticulocyte simulant. Red blood cell mimics, white blood cell mimics, platelet mimics, and nucleated red blood cell mimics can be added to prepare the required quality controls. Add erythrocyte simulant to 4×10 ¹² cells/L, adjust the reticulocyte ratio to 5% to 6%, and use a hematology analyzer based on nucleic acid fluorescence analysis principle for detection. The cluster is an erythrocyte mimetic, and the cell cluster on the right is a reticulocyte mimetic) only the pictures of the RET channel are provided.

It can be seen from Fig. 3 and Fig. 4 that the reticulocyte simulant prepared in Example 1 is far away from the position of the cell mass and the position of the red blood cell mass, which is conducive to the accurate identification of the instrument. During the actual preparation, the reaction conditions can also be controlled as required to adjust the position of the cell mass, such as the preparation method described in Example 2 and the test results shown in FIG. 4 .

It can be seen from the accompanying drawings 1-6 that the reticulocyte quality control substance prepared in each embodiment of the present invention is detected in a blood analyzer, and a reticulocyte mass having a clear boundary with the red blood cell mass is obtained. Therefore, the reticulocyte mimetic obtained by the method of the present invention can realize the quality control of reticulocytes based on the principle of nucleic acid fluorescent staining. The reticulocyte simulant obtained by the present invention can only add erythrocyte simulant to prepare for quality control of reticulocyte detection alone; erythrocyte simulant, leukocyte simulant, platelet simulant and nucleated erythrocyte simulant can also be added at the same time The whole blood quality control material containing all parameters is prepared for the quality control of all parameters detected by the blood analyzer.

Reagents used in the examples:

1. Anticoagulant: add 7.3 g of citric acid, 22.0 g of trisodium citrate, and 24.5 g of anhydrous glucose per liter of purified water. The osmotic pressure was adjusted to 400 ± 10 mOsm/kgH20 with NaCl.

2. Cell curing agent 1: per liter of purified water, add 8.0 mL of formaldehyde (37%), 1.5 mL of glutaraldehyde (25%), 0.125 g of dodecyl trimethyl ammonium chloride, and 1.5 g of cobalt chloride hexahydrate , Sodium chloride 9.0g.

3. Cell curing agent 2: per liter of purified water, add 6.0 mL of formaldehyde (37%), 1.0 mL of glutaraldehyde (25%), 0.125 g of dodecyl trimethyl ammonium chloride, and 1.0 g of cobalt chloride hexahydrate , Sodium chloride 9.0g.

3. Cell curing agent 3: add 5.0 mL of formaldehyde (37%), 1.5 mL of glutaraldehyde (25%), 0.04 g of dodecyl-β-d-maltoside, 5 g of anhydrous sodium sulfate, Sodium chloride 7.5g.

4. Cell curing agent 4: add 9.0 mL of formaldehyde (37%), 1.5 mL of glutaraldehyde (25%), 0.125 g of dodecyl trimethyl ammonium chloride, and 9.0 g of sodium chloride per liter of purified water.

5. Cell maintenance solution: penicillin 0.15g/L, gentamicin 0.15g/L, chloramphenicol 0.1g/L, glucose 2g/L, bovine serum albumin 20g/L, sodium chloride 6.3g/L, Sodium citrate 7.8g/L, citric acid 0.01g/L; the osmotic pressure of the cell maintenance solution is 305mOsm/kgH20, and the pH value is 7.32.

6. De-solidifying agent solution: bovine serum albumin 400g/L.

The instrument used in the test: XN-1000 blood analyzer with nucleic acid fluorescence analysis principle from Sysmex.

Beneficial effects:

1. The preparation method of the reticulocyte simulant provided by the present invention can be completed only by simple treatment, the preparation method is simple, the cost is low, and it is suitable for mass production.

2. The reticulocyte quality control substance provided by the present invention is prepared from animal blood and has high biological safety.

3. In the reticulocyte quality control material provided by the present invention, the red blood cell mass and the reticulocyte mass are completely separated, which is conducive to the stable identification of the instrument, and at the same time, the stability is high, and the bottle can be stable for more than 14 days after opening, and the validity period is 3 month or more.

What is disclosed above is only a preferred embodiment of the method for preparing a reticulocyte quality control substance based on a fluorescence principle of the present invention. Of course, it cannot limit the scope of rights of the present invention. Those of ordinary skill in the art can understand that the above embodiment can be realized. All or part of the process, and the equivalent changes made according to the claims of the present invention, still belong to the scope covered by the invention.

Claims (7)

1. a reticulocyte quality control substance preparation method of fluorescence principle, is characterized in that, comprises the following steps: Wash and separate anticoagulated blood to obtain anucleated red blood cells; using a cell solidifying agent to process and fix the anucleated red blood cells to obtain solidified red blood cells; The solidified red blood cells are treated with a de-solidifying agent to obtain a red blood cell product; The erythrocyte product is washed and stored in a cell maintenance solution to obtain a reticulocyte simulant. 2. the reticulocyte quality control substance preparation method of fluorescence principle as claimed in claim 1 is characterized in that, The specific method of washing and separating anticoagulated blood to obtain anucleated red blood cells is as follows: Mix anticoagulant with blood to get anticoagulated blood; The anticoagulated blood is washed and separated to obtain anucleated red blood cells. 3. the reticulocyte quality control substance preparation method of fluorescence principle as claimed in claim 1 is characterized in that, The described solidified red blood cells are treated with a de-solidifying agent, and the specific manner of obtaining the red blood cell product is as follows: After centrifuging the solidified red blood cells, remove the supernatant to obtain red blood cell pellets; A cell maintenance solution and a de-solidifying agent solution are added to the red blood cell pellet, and the solution is allowed to stand for 10 minutes to 1 hour to obtain a red blood cell product. 4. the reticulocyte quality control substance preparation method of fluorescence principle as claimed in claim 2 is characterized in that, The anticoagulant includes EDTA anticoagulant or citric acid anticoagulant; The cell fixative includes aldehyde cell fixative and base solution; The cell solidifying agent also includes any one of a cell spheroidizing agent and a surfactant; The cell curing agent also includes any one of a protein salting-out agent and a heavy metal salt, specifically including zinc sulfate, potassium sulfate, anhydrous sodium sulfate, ammonium sulfate, magnesium sulfate, zinc fluoride, barium chloride, and chlorine hexahydrate. Any one or more of cobalt; The surfactants include cationic surfactants, anionic surfactants and nonionic surfactants. 5. the reticulocyte quality control substance preparation method of fluorescence principle as claimed in claim 4 is characterized in that, Described cationic surfactant comprises quaternary ammonium salt type surfactant, specifically comprises any one in cetyl ammonium bromide, dodecyl trimethyl ammonium chloride; The anionic surfactants include sulfonate-based surfactants or sulfate-based surfactants, specifically including sodium dodecyl sulfate, sodium dodecylsulfonate and sodium dodecylbenzenesulfonate. any kind; The nonionic surfactant includes polyoxyethylene or polyol fatty acid ester surfactant, specifically including any one of polyethylene glycol and dodecyl-β-D-maltoside; The aldehyde cell fixative includes any one or more of formaldehyde, glutaraldehyde, o-phthalaldehyde and glyoxal; The bottom solution includes any one of sodium chloride solution, borate buffer, and phosphate buffer. 6. the reticulocyte quality control substance preparation method of fluorescence principle as claimed in claim 1 is characterized in that, The amount of the cell solidifying agent is 5-20 times the volume of the anucleated red blood cells. 7. the reticulocyte quality control substance preparation method of fluorescence principle as claimed in claim 3, is characterized in that, The osmotic pressure of the cell maintenance solution is 300±20mOsm/kgH20, the pH value is 7.3±0.1, and the cell maintenance solution includes antibiotics, polysaccharides, bovine serum albumin, sodium chloride, sodium citrate and citric acid; The antibiotics include any one or more of penicillins, aminoglycosides and amide antibiotics; The polysaccharide includes any one or more of sucrose, glucose, fructose, galactose, lactose, maltose, starch and dextrin.

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