CN112098181A - Raji staining solution used based on instrumental method and application thereof - Google Patents
Raji staining solution used based on instrumental method and application thereof Download PDFInfo
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- CN112098181A CN112098181A CN202010942995.9A CN202010942995A CN112098181A CN 112098181 A CN112098181 A CN 112098181A CN 202010942995 A CN202010942995 A CN 202010942995A CN 112098181 A CN112098181 A CN 112098181A
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- 239000012192 staining solution Substances 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 30
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 43
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 43
- 239000003599 detergent Substances 0.000 claims abstract description 40
- 239000000975 dye Substances 0.000 claims abstract description 34
- 239000000834 fixative Substances 0.000 claims abstract description 26
- 238000004043 dyeing Methods 0.000 claims abstract description 24
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims abstract description 20
- 229960000907 methylthioninium chloride Drugs 0.000 claims abstract description 20
- YOWZJZJLXUQHGF-UHFFFAOYSA-M [7-(dimethylamino)phenothiazin-3-ylidene]-dimethylazanium;dimethyl-[7-(methylamino)phenothiazin-3-ylidene]azanium;dichloride Chemical compound [Cl-].[Cl-].C1=CC(=[N+](C)C)C=C2SC3=CC(NC)=CC=C3N=C21.C1=CC(=[N+](C)C)C=C2SC3=CC(N(C)C)=CC=C3N=C21 YOWZJZJLXUQHGF-UHFFFAOYSA-M 0.000 claims abstract description 18
- SEACYXSIPDVVMV-UHFFFAOYSA-L eosin Y Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C([O-])=C(Br)C=C21 SEACYXSIPDVVMV-UHFFFAOYSA-L 0.000 claims abstract description 15
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000007864 aqueous solution Substances 0.000 claims abstract description 8
- 238000005406 washing Methods 0.000 claims abstract description 4
- 239000012153 distilled water Substances 0.000 claims description 66
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 66
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 36
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 32
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 32
- 238000010186 staining Methods 0.000 claims description 27
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 13
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 6
- 239000010452 phosphate Substances 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 5
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 5
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 5
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 abstract description 2
- 230000002349 favourable effect Effects 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract 4
- 230000003139 buffering effect Effects 0.000 abstract 2
- 239000002253 acid Substances 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 description 33
- 210000004027 cell Anatomy 0.000 description 32
- 210000004369 blood Anatomy 0.000 description 15
- 239000008280 blood Substances 0.000 description 15
- 238000002156 mixing Methods 0.000 description 13
- 230000000694 effects Effects 0.000 description 12
- 238000007447 staining method Methods 0.000 description 10
- 210000003855 cell nucleus Anatomy 0.000 description 5
- 210000000805 cytoplasm Anatomy 0.000 description 5
- 239000008187 granular material Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- DBZJJPROPLPMSN-UHFFFAOYSA-N bromoeosin Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Br)=C(O)C(Br)=C1OC1=C(Br)C(O)=C(Br)C=C21 DBZJJPROPLPMSN-UHFFFAOYSA-N 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 210000001772 blood platelet Anatomy 0.000 description 2
- 239000006153 eosin methylene blue Substances 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 201000004792 malaria Diseases 0.000 description 2
- 210000000440 neutrophil Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 210000003651 basophil Anatomy 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 239000001045 blue dye Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 230000002559 cytogenic effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000012333 histopathological diagnosis Methods 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Abstract
The invention is suitable for the technical field of medical examination, and provides a Rajjje's staining solution used based on an instrumental method and application thereof, wherein the Rajje's staining solution comprises: a cell fixative; the primary dye is an aqueous solution of eosin Y and potassium dihydrogen phosphate; the counterstain is an aqueous solution of methylene blue, azure II and potassium dihydrogen phosphate; and a buffer detergent. According to the dyeing liquid of the Raji's dyeing provided by the invention, the acid dyeing liquid eosin and the alkaline dyeing liquid azure II and methylene blue are respectively prepared into independent dyeing agents by selecting the azure II and the methylene blue as the secondary stainins, the steps of buffering and washing are combined, the steps of fixing, primary dyeing, secondary dyeing and buffering washing are used as independent steps during dyeing, a sample to be detected can be directly dried during instrument dyeing, the sample can be taken out for microscopic examination, the dyeing process is fast, errors caused by human factors can be avoided, and the dyeing liquid is favorable for popularization in clinical laboratories.
Description
Technical Field
The invention belongs to the technical field of medical examination, and particularly relates to a Rajbeck staining solution used based on an instrumental method and application thereof.
Background
The Reye dyeing method is a blood dyeing method invented by James Homer Reiter in 1902, and a classic Ruie dye consists of an acid eosin and a basic methylene blue dye, wherein the acid eosin and the basic methylene blue are mixed and become neutral eosin methylene blue after chemical action, and the neutral eosin methylene blue is contained after long-term oxidation. The three dyes respectively neutralize NH in cell nucleus and pulp3+And COO-And the like, to stain the nucleus and cytoplasm. Due to NH in cell nucleus and cell cytoplasm of different cell lines3+And COO-Different contents and different cell staining conditions, so that different blood cells can be easily distinguished by the method. The Ruhry staining method is mainly used for staining peripheral blood smear, urine sample and bone marrow, and is performed under an optical microscopeThe examination can assist in diagnosing clinical diseases such as leukemia, malaria and the like. But the method has 3 defects since the invention: (1) grinding is needed in preparation; (2) the dye can not be used immediately after being prepared, and can be dyed after being placed for more than half a year; (3) has better dyeing effect on cytoplasm and neutral granules, but has poorer dyeing effect on cell nucleus.
The Jiemsa color method is invented by German chemist and bacteriologist Gustav Giemsa, is a nucleic acid staining agent, and the staining solution consists of eosin and azure, and the staining principle is the same as that of the Ruhry staining method, and can be used for cytogenetics and histopathological diagnosis of malaria and other parasites. However, this method has a good effect on staining nuclei but a poor effect on staining cytoplasm.
The Rujje's staining method is a method for satisfying the staining effect of cell nucleus and cytoplasm by using the advantages of the Rujje's staining method and the Jimsa's staining method, and the method can be applied to the Rujje's staining method and the Jimsa's staining method. Currently, the development of the rebaudius dyeing method is rapid, a plurality of formulas and operation methods of the rebaudius dyeing method are presented, and three methods, namely a rebaudius complex dyeing method, a 30-second quick single dyeing method and a quick dyeing method, are listed in national clinical examination operation regulations (4 th edition), and the defects that a dye solution needs to be stored for a period of time and can be used, and the newly prepared dye solution has a poor dyeing effect.
Disclosure of Invention
The embodiment of the invention aims to provide a Ray Ji's staining solution used based on an instrumental method, and aims to solve the problems in the background art.
The embodiment of the invention is realized by the method, and the method is based on the instrument method and used for the dyeing liquor of the Rajbec, which comprises the following steps:
a cell fixative;
the primary dye is an aqueous solution of eosin Y and potassium dihydrogen phosphate;
the counterstain is an aqueous solution of methylene blue, azure II and potassium dihydrogen phosphate; and
a buffer detergent.
As a preferred embodiment of the present invention, the cell fixative is methanol.
As another preferred embodiment of the present invention, every 500mL of the primary dye comprises the following components: 0.5-0.9 g of eosin Y, 4-6 g of monopotassium phosphate and the balance of distilled water.
As another preferred embodiment of the present invention, every 500mL of the counterstain comprises the following components: 0.7-1 g of methylene blue, 0.7-1 g of azure II, 2-4 g of monopotassium phosphate and the balance of distilled water.
As another preferred version of the embodiment of the present invention, the buffer detergent is an aqueous solution of phosphate.
In another preferred embodiment of the present invention, the phosphate is at least one of disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, and potassium dihydrogen phosphate.
As another preferable mode of the embodiment of the present invention, the phosphate is disodium hydrogen phosphate and potassium dihydrogen phosphate.
As another preferred embodiment of the present invention, the buffer detergent comprises the following components per 500 mL: 0.2 to 0.4g of disodium hydrogen phosphate, 0.1 to 0.5g of monopotassium phosphate and the balance of distilled water.
Another object of the present invention is to provide an application of the above-mentioned staining solution for rebalancing cells.
As another preferable aspect of the embodiment of the present invention, the application includes the steps of:
fixing a sample to be stained by using a cell fixing agent;
sequentially dyeing the fixed sample to be dyed with a primary dyeing agent and a secondary dyeing agent;
and washing the dyed sample with a buffer detergent.
The invention provides a Rui Ji's staining solution used based on an instrumental method, and aims to solve the inherent defects of longer preparation time and other methods existing in the traditional manual Rui Ji's staining method, the Ji's staining method and the Rui Ji's-Ji's staining method and the problem of interference of human factors on staining quality. Specifically, the acid eosin, the alkaline methylene blue and the azure II are respectively prepared into solutions, so that the staining solution can be used immediately after preparation, and the cell morphology can be identified under the microscope by the slide stained by the instrument, the whole slide staining process is about 3 minutes, errors caused by human factors can be avoided, and the method is favorable for popularization in clinical laboratories.
Drawings
FIG. 1 is a graph showing the staining effect of the staining solution of Rajbech's stain on clinical blood smears, which is provided in example 1 of the present invention.
FIG. 2 is a graph showing the staining effect of the staining solution of the invention provided in example 1 on another group of clinical blood smears.
FIG. 3 is a graph showing the staining effect of the staining solution of the invention provided in example 1 on another group of clinical blood smears.
FIG. 4 is a graph showing the staining effect of the staining solution of the invention provided in example 1 on another group of clinical blood smears.
FIG. 5 is a graph showing the staining effect of the staining solution of the invention provided in example 1 on another group of clinical blood smears.
FIG. 6 is a graph showing the staining effect of the staining solution of the invention provided in example 1 on another group of clinical blood smears.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.8g of eosin Y, 5g of monopotassium phosphate and 400mL of distilled water, and then using the distilled water to fix the volume to 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 1g of methylene blue, 1g of azure II, 2.5g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.3g of disodium hydrogen phosphate, 0.2g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Example 2
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.5g of eosin Y, 4g of monopotassium phosphate and 400mL of distilled water, and then using the distilled water to fix the volume to 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.7g of methylene blue, 0.7g of azure II, 2g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.2g of disodium hydrogen phosphate, 0.1g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Example 3
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.9g of eosin Y, 6g of monopotassium phosphate and 400mL of distilled water, and then using the distilled water to fix the volume to 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 1g of methylene blue, 1g of azure II, 4g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.4g of disodium hydrogen phosphate, 0.5g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Example 4
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.6g of eosin Y, 4.5g of monopotassium phosphate and 400mL of distilled water, and then adding distilled water to a constant volume of 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.8g of methylene blue, 0.8g of azure II, 2.5g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: and uniformly mixing 0.3g of dipotassium hydrogen phosphate, 0.3g of sodium dihydrogen phosphate and 400mL of distilled water, and then adding the distilled water to a constant volume of 500mL to obtain the buffer detergent.
Example 5
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.8g of eosin Y, 5.5g of monopotassium phosphate and 400mL of distilled water, and then adding distilled water to a constant volume of 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.9g of methylene blue, 0.9g of azure II, 3.5g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.1g of disodium hydrogen phosphate, 0.2g of potassium dihydrogen phosphate, 0.1g of dipotassium hydrogen phosphate, 0.2g of sodium dihydrogen phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Example 6
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.7g of eosin Y, 5g of monopotassium phosphate and 400mL of distilled water, and then using the distilled water to fix the volume to 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.85g of methylene blue, 0.85g of azure II, 3g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: and uniformly mixing 0.8g of monopotassium phosphate with 400mL of distilled water, and then diluting the mixture to 500mL by using the distilled water to obtain the buffer detergent.
Example 7
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.8g of eosin Y, 4.2g of monopotassium phosphate and 400mL of distilled water, and then adding distilled water to a constant volume of 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.75g of methylene blue, 0.95g of azure II, 3g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: and uniformly mixing 0.6g of disodium hydrogen phosphate with 400mL of distilled water, and then adding distilled water to a constant volume of 500mL to obtain the buffer detergent.
Example 8
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.7g of eosin Y, 5.3g of monopotassium phosphate and 400mL of distilled water, and then adding distilled water to a constant volume of 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.9g of methylene blue, 0.8g of azure II, 3.3g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.3g of disodium hydrogen phosphate, 0.1g of monopotassium phosphate, 0.1g of dipotassium hydrogen phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Example 9
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.7g of eosin Y, 5g of monopotassium phosphate and 400mL of distilled water, and then using the distilled water to fix the volume to 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.8g of methylene blue, 0.8g of azure II, 2.4g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.3g of disodium hydrogen phosphate, 0.2g of dipotassium hydrogen phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Example 10
This example provides an instrumental-based staining solution of RAIL, which includes a cell fixative, a primary stain, a secondary stain, and a buffer detergent.
Wherein the cell fixative is methanol.
The preparation method of the primary dye comprises the following steps: uniformly mixing 0.6g of eosin Y, 5g of monopotassium phosphate and 400mL of distilled water, and then using the distilled water to fix the volume to 500mL to obtain the primary dye.
The preparation method of the counterstain comprises the following steps: 0.8g of methylene blue, 0.8g of azure II, 3.4g of monopotassium phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to 500mL by using the distilled water, so that the counterstain is obtained.
The preparation method of the buffer detergent comprises the following steps: 0.3g of monopotassium phosphate, 0.3g of sodium dihydrogen phosphate and 400mL of distilled water are mixed uniformly, and then the volume is determined to be 500mL by using the distilled water, so that the buffer detergent is obtained.
Experimental example:
selecting 10 blood routine detection specimens given clinically, performing staining microscopic examination after production, and performing image acquisition by an image acquisition system. Wherein the dyeing step comprises the following steps: after the blood smear is naturally dried, the blood smear is placed in a HaoXin staining machine, the blood smear is stained according to a blood staining mode, and the staining solution of the Ruijie staining solution provided by the embodiment 1 is used for staining, specifically, the blood smear is firstly fixed for 1.5 minutes by using a cell fixative, then is sequentially stained for 15 seconds by using an initial staining agent and a counterstaining agent for 15 seconds, and then is treated for 1 minute by using a buffer detergent (the whole staining process only needs about 3 minutes), and after the instrument is stopped, the slide can be taken out for microscopic examination. The microscopic staining effect graphs of 6 clinical blood smears are respectively shown in figures 1-6. As can be seen from the figure, after the clinical blood smear is stained by the RAIL staining solution provided by the embodiment of the invention, lymphocytes, neutrophils, erythrocytes, platelets and the like which meet the staining requirements can be seen in microscopic examination.
The cell staining solution provided by the invention is used for staining the cells, and a clear cell staining result can be obtained: after staining, the cell nucleus is purple red, the lymphocyte plasma is grayish blue, the neutrophil granules are light purple, the eosinophil granules are red to reddish brown, the basophil granules are purple black to black, the platelet is purple, and the red cells are red, so that the feasibility of the Reibec's staining solution can be proved.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A Rajie's staining solution used based on an instrumental method, comprising:
a cell fixative;
the primary dye is an aqueous solution of eosin Y and potassium dihydrogen phosphate;
the counterstain is an aqueous solution of methylene blue, azure II and potassium dihydrogen phosphate; and
a buffer detergent.
2. The apparatus-based staining solution of RAIL according to claim 1, wherein the cell fixative is methanol.
3. The apparatus-based staining solution of RAIL according to claim 1, wherein the primary stain comprises the following components per 500 mL: 0.5-0.9 g of eosin Y, 4-6 g of monopotassium phosphate and the balance of distilled water.
4. The apparatus-based RAIL staining solution as claimed in claim 1, wherein the counterstain comprises the following components per 500 mL: 0.7-1 g of methylene blue, 0.7-1 g of azure II, 2-4 g of monopotassium phosphate and the balance of distilled water.
5. The apparatus-based staining solution of RAIL according to claim 1, wherein the buffer detergent is an aqueous solution of phosphate.
6. The staining solution of RAPIS for use according to claim 5, wherein the phosphate is at least one of disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, and potassium dihydrogen phosphate.
7. A staining solution according to claim 5 or 6, wherein the phosphate is disodium hydrogen phosphate and potassium dihydrogen phosphate.
8. The apparatus-based staining solution of RAIL according to claim 7, wherein the buffer detergent comprises the following components per 500 mL: 0.2 to 0.4g of disodium hydrogen phosphate, 0.1 to 0.5g of monopotassium phosphate and the balance of distilled water.
9. Use of the RAIL-ABM staining solution of any one of claims 1 to 8 for cell staining.
10. Use according to claim 9, characterized in that it comprises the following steps:
fixing a sample to be stained by using a cell fixing agent;
sequentially dyeing the fixed sample to be dyed with a primary dyeing agent and a secondary dyeing agent;
and washing the dyed sample with a buffer detergent.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4741898A (en) * | 1985-04-01 | 1988-05-03 | Fisher Scientific Company | Stabilized stain composition |
| JP2002350431A (en) * | 2001-05-23 | 2002-12-04 | Nippon Koden Corp | Decoloring method and device of stained specimen |
| CN101587039A (en) * | 2009-07-07 | 2009-11-25 | 中国人民解放军第三军医大学第二附属医院 | A kind of blood cell stain |
| CN109406246A (en) * | 2018-10-25 | 2019-03-01 | 潍坊市康华生物技术有限公司 | A kind of sperm morphological rapid dyeing reagent and its colouring method |
-
2020
- 2020-09-09 CN CN202010942995.9A patent/CN112098181A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4741898A (en) * | 1985-04-01 | 1988-05-03 | Fisher Scientific Company | Stabilized stain composition |
| JP2002350431A (en) * | 2001-05-23 | 2002-12-04 | Nippon Koden Corp | Decoloring method and device of stained specimen |
| CN101587039A (en) * | 2009-07-07 | 2009-11-25 | 中国人民解放军第三军医大学第二附属医院 | A kind of blood cell stain |
| CN109406246A (en) * | 2018-10-25 | 2019-03-01 | 潍坊市康华生物技术有限公司 | A kind of sperm morphological rapid dyeing reagent and its colouring method |
Non-Patent Citations (1)
| Title |
|---|
| 李乃津等: "莓形体的两种快速染色法", 《家畜传染病》 * |
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