CN105074422B - 用于染色和样品处理的方法和组合物 - Google Patents
用于染色和样品处理的方法和组合物 Download PDFInfo
- Publication number
- CN105074422B CN105074422B CN201480013419.9A CN201480013419A CN105074422B CN 105074422 B CN105074422 B CN 105074422B CN 201480013419 A CN201480013419 A CN 201480013419A CN 105074422 B CN105074422 B CN 105074422B
- Authority
- CN
- China
- Prior art keywords
- contrast agent
- enough
- sample
- amount
- particle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 146
- 238000004043 dyeing Methods 0.000 title claims abstract description 128
- 238000000034 method Methods 0.000 title claims abstract description 56
- 239000002245 particle Substances 0.000 claims abstract description 174
- 239000002872 contrast media Substances 0.000 claims abstract description 141
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 58
- 239000013078 crystal Substances 0.000 claims abstract description 52
- NZYCYASKVWSANA-UHFFFAOYSA-M new methylene blue Chemical compound [Cl-].CCNC1=C(C)C=C2N=C(C=C(C(NCC)=C3)C)C3=[S+]C2=C1 NZYCYASKVWSANA-UHFFFAOYSA-M 0.000 claims abstract description 47
- 239000000834 fixative Substances 0.000 claims abstract description 30
- 230000000740 bleeding effect Effects 0.000 claims abstract description 28
- HGUVPEBGCAVWID-KETMJRJWSA-N 7-O-(beta-D-glucosyl)isovitexin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1O)[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)=CC2=C1C(=O)C=C(C=1C=CC(O)=CC=1)O2 HGUVPEBGCAVWID-KETMJRJWSA-N 0.000 claims abstract description 21
- HGUVPEBGCAVWID-UHFFFAOYSA-N saponarin Natural products OC1C(O)C(O)C(CO)OC1OC(C(=C1O)C2C(C(O)C(O)C(CO)O2)O)=CC2=C1C(=O)C=C(C=1C=CC(O)=CC=1)O2 HGUVPEBGCAVWID-UHFFFAOYSA-N 0.000 claims abstract description 21
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000003384 imaging method Methods 0.000 claims description 25
- 238000004458 analytical method Methods 0.000 claims description 23
- 230000017531 blood circulation Effects 0.000 claims description 17
- 239000000872 buffer Substances 0.000 claims description 6
- 238000010438 heat treatment Methods 0.000 claims description 4
- 238000004040 coloring Methods 0.000 abstract description 3
- 239000000523 sample Substances 0.000 description 119
- 210000004027 cell Anatomy 0.000 description 73
- 210000000265 leukocyte Anatomy 0.000 description 27
- 239000008280 blood Substances 0.000 description 22
- 210000004369 blood Anatomy 0.000 description 21
- 239000003086 colorant Substances 0.000 description 20
- 239000012530 fluid Substances 0.000 description 20
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 18
- 210000003743 erythrocyte Anatomy 0.000 description 17
- 238000010586 diagram Methods 0.000 description 16
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
- 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 description 15
- 239000000975 dye Substances 0.000 description 14
- 238000010186 staining Methods 0.000 description 14
- 210000001772 blood platelet Anatomy 0.000 description 13
- OARRHUQTFTUEOS-UHFFFAOYSA-N safranin Chemical compound [Cl-].C=12C=C(N)C(C)=CC2=NC2=CC(C)=C(N)C=C2[N+]=1C1=CC=CC=C1 OARRHUQTFTUEOS-UHFFFAOYSA-N 0.000 description 13
- 239000007788 liquid Substances 0.000 description 12
- DWCZIOOZPIDHAB-UHFFFAOYSA-L methyl green Chemical compound [Cl-].[Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC(=CC=1)[N+](C)(C)C)=C1C=CC(=[N+](C)C)C=C1 DWCZIOOZPIDHAB-UHFFFAOYSA-L 0.000 description 12
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 11
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 11
- 239000002953 phosphate buffered saline Substances 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- HBXWUCXDUUJDRB-UHFFFAOYSA-N 1-octadecoxyoctadecane Chemical compound CCCCCCCCCCCCCCCCCCOCCCCCCCCCCCCCCCCCC HBXWUCXDUUJDRB-UHFFFAOYSA-N 0.000 description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 230000000007 visual effect Effects 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- 210000003979 eosinophil Anatomy 0.000 description 7
- 210000003714 granulocyte Anatomy 0.000 description 7
- 230000003287 optical effect Effects 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 238000002601 radiography Methods 0.000 description 7
- FFJCNSLCJOQHKM-CLFAGFIQSA-N (z)-1-[(z)-octadec-9-enoxy]octadec-9-ene Chemical compound CCCCCCCC\C=C/CCCCCCCCOCCCCCCCC\C=C/CCCCCCCC FFJCNSLCJOQHKM-CLFAGFIQSA-N 0.000 description 6
- -1 Dimethyl diaminophenazine chloride Chemical compound 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 238000004820 blood count Methods 0.000 description 6
- 210000000440 neutrophil Anatomy 0.000 description 6
- 210000003924 normoblast Anatomy 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 239000008187 granular material Substances 0.000 description 5
- 210000004698 lymphocyte Anatomy 0.000 description 5
- 210000001616 monocyte Anatomy 0.000 description 5
- 239000002736 nonionic surfactant Substances 0.000 description 5
- 230000009467 reduction Effects 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- CMCBDXRRFKYBDG-UHFFFAOYSA-N 1-dodecoxydodecane Chemical compound CCCCCCCCCCCCOCCCCCCCCCCCC CMCBDXRRFKYBDG-UHFFFAOYSA-N 0.000 description 4
- 125000001931 aliphatic group Chemical group 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- KRVSOGSZCMJSLX-UHFFFAOYSA-L chromic acid Substances O[Cr](O)(=O)=O KRVSOGSZCMJSLX-UHFFFAOYSA-L 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000002708 enhancing effect Effects 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 230000003834 intracellular effect Effects 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- FDCJDKXCCYFOCV-UHFFFAOYSA-N 1-hexadecoxyhexadecane Chemical compound CCCCCCCCCCCCCCCCOCCCCCCCCCCCCCCCC FDCJDKXCCYFOCV-UHFFFAOYSA-N 0.000 description 3
- 102000001554 Hemoglobins Human genes 0.000 description 3
- 108010054147 Hemoglobins Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000004218 Orcein Substances 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 210000000601 blood cell Anatomy 0.000 description 3
- 238000002296 dynamic light scattering Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- VKYKSIONXSXAKP-UHFFFAOYSA-N hexamethylenetetramine Chemical compound C1N(C2)CN3CN1CN2C3 VKYKSIONXSXAKP-UHFFFAOYSA-N 0.000 description 3
- 239000000416 hydrocolloid Substances 0.000 description 3
- 230000008595 infiltration Effects 0.000 description 3
- 238000001764 infiltration Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000000366 juvenile effect Effects 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 210000003887 myelocyte Anatomy 0.000 description 3
- 235000019248 orcein Nutrition 0.000 description 3
- 229910000489 osmium tetroxide Inorganic materials 0.000 description 3
- 239000011886 peripheral blood Substances 0.000 description 3
- 230000035699 permeability Effects 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 210000004765 promyelocyte Anatomy 0.000 description 3
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 3
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 210000000130 stem cell Anatomy 0.000 description 3
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 3
- 239000002888 zwitterionic surfactant Substances 0.000 description 3
- KJCVRFUGPWSIIH-UHFFFAOYSA-N 1-naphthol Chemical compound C1=CC=C2C(O)=CC=CC2=C1 KJCVRFUGPWSIIH-UHFFFAOYSA-N 0.000 description 2
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- JUQPZRLQQYSMEQ-UHFFFAOYSA-N CI Basic red 9 Chemical compound [Cl-].C1=CC(N)=CC=C1C(C=1C=CC(N)=CC=1)=C1C=CC(=[NH2+])C=C1 JUQPZRLQQYSMEQ-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 229920001503 Glucan Polymers 0.000 description 2
- 241001062009 Indigofera Species 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 240000003946 Saponaria officinalis Species 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- WLKAMFOFXYCYDK-UHFFFAOYSA-N [5-amino-4-[[3-[(2-amino-4-azaniumyl-5-methylphenyl)diazenyl]-4-methylphenyl]diazenyl]-2-methylphenyl]azanium;dichloride Chemical compound [Cl-].[Cl-].CC1=CC=C(N=NC=2C(=CC([NH3+])=C(C)C=2)N)C=C1N=NC1=CC(C)=C([NH3+])C=C1N WLKAMFOFXYCYDK-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 150000008378 aryl ethers Chemical class 0.000 description 2
- 239000000987 azo dye Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 235000012730 carminic acid Nutrition 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000005336 cracking Methods 0.000 description 2
- ZXJXZNDDNMQXFV-UHFFFAOYSA-M crystal violet Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1[C+](C=1C=CC(=CC=1)N(C)C)C1=CC=C(N(C)C)C=C1 ZXJXZNDDNMQXFV-UHFFFAOYSA-M 0.000 description 2
- DTPCFIHYWYONMD-UHFFFAOYSA-N decaethylene glycol Polymers OCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO DTPCFIHYWYONMD-UHFFFAOYSA-N 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 description 2
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 2
- RFKJHQXSLBUONF-UHFFFAOYSA-N methyl blue free acid Chemical compound C1=CC(S(=O)(=O)O)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=NC=2C=CC(=CC=2)S(O)(=O)=O)C=2C=CC(NC=3C=CC(=CC=3)S(O)(=O)=O)=CC=2)C=C1 RFKJHQXSLBUONF-UHFFFAOYSA-N 0.000 description 2
- 210000000633 nuclear envelope Anatomy 0.000 description 2
- 239000007800 oxidant agent Substances 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000011164 primary particle Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- CXZRDVVUVDYSCQ-UHFFFAOYSA-M pyronin B Chemical compound [Cl-].C1=CC(=[N+](CC)CC)C=C2OC3=CC(N(CC)CC)=CC=C3C=C21 CXZRDVVUVDYSCQ-UHFFFAOYSA-M 0.000 description 2
- 238000009738 saturating Methods 0.000 description 2
- 239000011163 secondary particle Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000012128 staining reagent Substances 0.000 description 2
- 108700024661 strong silver Proteins 0.000 description 2
- 210000004895 subcellular structure Anatomy 0.000 description 2
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- LNOBZXNCABUBKK-UHFFFAOYSA-N 2,3,5-triphenyltetrazolium Chemical compound C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 LNOBZXNCABUBKK-UHFFFAOYSA-N 0.000 description 1
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 1
- DUIOKRXOKLLURE-UHFFFAOYSA-N 2-octylphenol Chemical compound CCCCCCCCC1=CC=CC=C1O DUIOKRXOKLLURE-UHFFFAOYSA-N 0.000 description 1
- OALHHIHQOFIMEF-UHFFFAOYSA-N 3',6'-dihydroxy-2',4',5',7'-tetraiodo-3h-spiro[2-benzofuran-1,9'-xanthene]-3-one Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 OALHHIHQOFIMEF-UHFFFAOYSA-N 0.000 description 1
- JKYKXTRKURYNGW-UHFFFAOYSA-N 3,4-dihydroxy-9,10-dioxo-9,10-dihydroanthracene-2-sulfonic acid Chemical compound O=C1C2=CC=CC=C2C(=O)C2=C1C(O)=C(O)C(S(O)(=O)=O)=C2 JKYKXTRKURYNGW-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- GUQQBLRVXOUDTN-XOHPMCGNSA-N 3-[dimethyl-[3-[[(4r)-4-[(3r,5s,7r,8r,9s,10s,12s,13r,14s,17r)-3,7,12-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]propyl]azaniumyl]-2-hydroxypropane-1-sulfonate Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCC[N+](C)(C)CC(O)CS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 GUQQBLRVXOUDTN-XOHPMCGNSA-N 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- OLQIKGSZDTXODA-UHFFFAOYSA-N 4-[3-(4-hydroxy-2-methylphenyl)-1,1-dioxo-2,1$l^{6}-benzoxathiol-3-yl]-3-methylphenol Chemical compound CC1=CC(O)=CC=C1C1(C=2C(=CC(O)=CC=2)C)C2=CC=CC=C2S(=O)(=O)O1 OLQIKGSZDTXODA-UHFFFAOYSA-N 0.000 description 1
- ALJHHTHBYJROOG-UHFFFAOYSA-N 7-(dimethylamino)phenothiazin-3-one Chemical compound C1=CC(=O)C=C2SC3=CC(N(C)C)=CC=C3N=C21 ALJHHTHBYJROOG-UHFFFAOYSA-N 0.000 description 1
- MPVDXIMFBOLMNW-ISLYRVAYSA-N 7-hydroxy-8-[(E)-phenyldiazenyl]naphthalene-1,3-disulfonic acid Chemical compound OC1=CC=C2C=C(S(O)(=O)=O)C=C(S(O)(=O)=O)C2=C1\N=N\C1=CC=CC=C1 MPVDXIMFBOLMNW-ISLYRVAYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-M Acrylate Chemical compound [O-]C(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-M 0.000 description 1
- SGHZXLIDFTYFHQ-UHFFFAOYSA-L Brilliant Blue Chemical compound [Na+].[Na+].C=1C=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C(=CC=CC=2)S([O-])(=O)=O)C=CC=1N(CC)CC1=CC=CC(S([O-])(=O)=O)=C1 SGHZXLIDFTYFHQ-UHFFFAOYSA-L 0.000 description 1
- DNPQJRSJLTVCCU-UHFFFAOYSA-N C(CCCCCCCCC)[N+](C)(C)OS(=O)(=O)CCC Chemical class C(CCCCCCCCC)[N+](C)(C)OS(=O)(=O)CCC DNPQJRSJLTVCCU-UHFFFAOYSA-N 0.000 description 1
- GAWIXWVDTYZWAW-UHFFFAOYSA-N C[CH]O Chemical group C[CH]O GAWIXWVDTYZWAW-UHFFFAOYSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Chinese gallotannin Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 244000303965 Cyamopsis psoralioides Species 0.000 description 1
- 102000018832 Cytochromes Human genes 0.000 description 1
- 108010052832 Cytochromes Proteins 0.000 description 1
- 239000004214 Fast Green FCF Substances 0.000 description 1
- RZSYLLSAWYUBPE-UHFFFAOYSA-L Fast green FCF Chemical compound [Na+].[Na+].C=1C=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C(=CC(O)=CC=2)S([O-])(=O)=O)C=CC=1N(CC)CC1=CC=CC(S([O-])(=O)=O)=C1 RZSYLLSAWYUBPE-UHFFFAOYSA-L 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 238000002738 Giemsa staining Methods 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 241000521257 Hydrops Species 0.000 description 1
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 1
- XXACTDWGHQXLGW-UHFFFAOYSA-M Janus Green B chloride Chemical compound [Cl-].C12=CC(N(CC)CC)=CC=C2N=C2C=CC(\N=N\C=3C=CC(=CC=3)N(C)C)=CC2=[N+]1C1=CC=CC=C1 XXACTDWGHQXLGW-UHFFFAOYSA-M 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 229920000161 Locust bean gum Polymers 0.000 description 1
- 229910001051 Magnalium Inorganic materials 0.000 description 1
- WWKGVZASJYXZKN-UHFFFAOYSA-N Methyl violet 2B Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC(N)=CC=1)=C1C=CC(=[N+](C)C)C=C1 WWKGVZASJYXZKN-UHFFFAOYSA-N 0.000 description 1
- 241000625836 Ochrolechia Species 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 1
- 240000008254 Rosa chinensis Species 0.000 description 1
- 235000000664 Rosa chinensis Nutrition 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 240000002825 Solanum vestissimum Species 0.000 description 1
- 235000018259 Solanum vestissimum Nutrition 0.000 description 1
- 108010076830 Thionins Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- JCGCKSUCGVTMNB-UHFFFAOYSA-N acetic acid;formaldehyde Chemical compound O=C.CC(O)=O JCGCKSUCGVTMNB-UHFFFAOYSA-N 0.000 description 1
- HCNZRGXMIOIZCH-UHFFFAOYSA-N acetic acid;phenol Chemical compound CC(O)=O.CC(O)=O.OC1=CC=CC=C1 HCNZRGXMIOIZCH-UHFFFAOYSA-N 0.000 description 1
- RZUBARUFLYGOGC-MTHOTQAESA-L acid fuchsin Chemical compound [Na+].[Na+].[O-]S(=O)(=O)C1=C(N)C(C)=CC(C(=C\2C=C(C(=[NH2+])C=C/2)S([O-])(=O)=O)\C=2C=C(C(N)=CC=2)S([O-])(=O)=O)=C1 RZUBARUFLYGOGC-MTHOTQAESA-L 0.000 description 1
- DGOBMKYRQHEFGQ-UHFFFAOYSA-L acid green 5 Chemical group [Na+].[Na+].C=1C=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)S([O-])(=O)=O)C=CC=1N(CC)CC1=CC=CC(S([O-])(=O)=O)=C1 DGOBMKYRQHEFGQ-UHFFFAOYSA-L 0.000 description 1
- CQPFMGBJSMSXLP-UHFFFAOYSA-M acid orange 7 Chemical compound [Na+].OC1=CC=C2C=CC=CC2=C1N=NC1=CC=C(S([O-])(=O)=O)C=C1 CQPFMGBJSMSXLP-UHFFFAOYSA-M 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000005215 alkyl ethers Chemical class 0.000 description 1
- 235000011126 aluminium potassium sulphate Nutrition 0.000 description 1
- WLDHEUZGFKACJH-UHFFFAOYSA-K amaranth Chemical compound [Na+].[Na+].[Na+].C12=CC=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(O)=C1N=NC1=CC=C(S([O-])(=O)=O)C2=CC=CC=C12 WLDHEUZGFKACJH-UHFFFAOYSA-K 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 210000004381 amniotic fluid Anatomy 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000003567 ascitic fluid Anatomy 0.000 description 1
- KSCQDDRPFHTIRL-UHFFFAOYSA-N auramine O Chemical compound [H+].[Cl-].C1=CC(N(C)C)=CC=C1C(=N)C1=CC=C(N(C)C)C=C1 KSCQDDRPFHTIRL-UHFFFAOYSA-N 0.000 description 1
- 229940052223 basic fuchsin Drugs 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004159 blood analysis Methods 0.000 description 1
- 239000001045 blue dye Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 238000009583 bone marrow aspiration Methods 0.000 description 1
- 235000012745 brilliant blue FCF Nutrition 0.000 description 1
- 239000004161 brilliant blue FCF Substances 0.000 description 1
- 229940055580 brilliant blue fcf Drugs 0.000 description 1
- 229960001506 brilliant green Drugs 0.000 description 1
- HXCILVUBKWANLN-UHFFFAOYSA-N brilliant green cation Chemical compound C1=CC(N(CC)CC)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](CC)CC)C=C1 HXCILVUBKWANLN-UHFFFAOYSA-N 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000012482 calibration solution Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 239000000679 carrageenan Substances 0.000 description 1
- 235000010418 carrageenan Nutrition 0.000 description 1
- 229920001525 carrageenan Polymers 0.000 description 1
- 229940113118 carrageenan Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 210000004081 cilia Anatomy 0.000 description 1
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical group [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 description 1
- 125000000853 cresyl group Chemical group C1(=CC=C(C=C1)C)* 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000002559 cytogenic effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- SOROIESOUPGGFO-UHFFFAOYSA-N diazolidinylurea Chemical compound OCNC(=O)N(CO)C1N(CO)C(=O)N(CO)C1=O SOROIESOUPGGFO-UHFFFAOYSA-N 0.000 description 1
- 229960001083 diazolidinylurea Drugs 0.000 description 1
- GKJMOBYTOXEHKJ-UHFFFAOYSA-N dihydroxymethylurea Chemical compound NC(=O)NC(O)O GKJMOBYTOXEHKJ-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000019240 fast green FCF Nutrition 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- AWJWCTOOIBYHON-UHFFFAOYSA-N furo[3,4-b]pyrazine-5,7-dione Chemical compound C1=CN=C2C(=O)OC(=O)C2=N1 AWJWCTOOIBYHON-UHFFFAOYSA-N 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 229960001235 gentian violet Drugs 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- 125000005908 glyceryl ester group Chemical group 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 229940015043 glyoxal Drugs 0.000 description 1
- 239000001046 green dye Substances 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 238000005534 hematocrit Methods 0.000 description 1
- 235000010299 hexamethylene tetramine Nutrition 0.000 description 1
- 239000004312 hexamethylene tetramine Substances 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 125000002632 imidazolidinyl group Chemical group 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- KHLVKKOJDHCJMG-QDBORUFSSA-L indigo carmine Chemical compound [Na+].[Na+].N/1C2=CC=C(S([O-])(=O)=O)C=C2C(=O)C\1=C1/NC2=CC=C(S(=O)(=O)[O-])C=C2C1=O KHLVKKOJDHCJMG-QDBORUFSSA-L 0.000 description 1
- 229960003988 indigo carmine Drugs 0.000 description 1
- 235000012738 indigotine Nutrition 0.000 description 1
- 239000004179 indigotine Substances 0.000 description 1
- 210000005061 intracellular organelle Anatomy 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000013010 irrigating solution Substances 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 239000000711 locust bean gum Substances 0.000 description 1
- 235000010420 locust bean gum Nutrition 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 229940107698 malachite green Drugs 0.000 description 1
- FDZZZRQASAIRJF-UHFFFAOYSA-M malachite green Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](C)C)C=C1 FDZZZRQASAIRJF-UHFFFAOYSA-M 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000010339 medical test Methods 0.000 description 1
- 229940008716 mercurochrome Drugs 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229960004011 methenamine Drugs 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229940012189 methyl orange Drugs 0.000 description 1
- STZCRXQWRGQSJD-GEEYTBSJSA-M methyl orange Chemical compound [Na+].C1=CC(N(C)C)=CC=C1\N=N\C1=CC=C(S([O-])(=O)=O)C=C1 STZCRXQWRGQSJD-GEEYTBSJSA-M 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 108700019599 monomethylolglycine Proteins 0.000 description 1
- 210000000272 myelencephalon Anatomy 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- YWFWDNVOPHGWMX-UHFFFAOYSA-N n,n-dimethyldodecan-1-amine Chemical compound CCCCCCCCCCCCN(C)C YWFWDNVOPHGWMX-UHFFFAOYSA-N 0.000 description 1
- 210000004493 neutrocyte Anatomy 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- SNQQPOLDUKLAAF-UHFFFAOYSA-N nonylphenol Chemical compound CCCCCCCCCC1=CC=CC=C1O SNQQPOLDUKLAAF-UHFFFAOYSA-N 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000012285 osmium tetroxide Substances 0.000 description 1
- 230000000065 osmolyte Effects 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- QUBQYFYWUJJAAK-UHFFFAOYSA-N oxymethurea Chemical compound OCNC(=O)NCO QUBQYFYWUJJAAK-UHFFFAOYSA-N 0.000 description 1
- 229950005308 oxymethurea Drugs 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 229940075930 picrate Drugs 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-M picrate anion Chemical compound [O-]C1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-M 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- 229940050271 potassium alum Drugs 0.000 description 1
- GRLPQNLYRHEGIJ-UHFFFAOYSA-J potassium aluminium sulfate Chemical compound [Al+3].[K+].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O GRLPQNLYRHEGIJ-UHFFFAOYSA-J 0.000 description 1
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 239000004302 potassium sorbate Substances 0.000 description 1
- 235000010241 potassium sorbate Nutrition 0.000 description 1
- 229940069338 potassium sorbate Drugs 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- YBBJKCMMCRQZMA-UHFFFAOYSA-N pyrithione Chemical compound ON1C=CC=CC1=S YBBJKCMMCRQZMA-UHFFFAOYSA-N 0.000 description 1
- INCIMLINXXICKS-UHFFFAOYSA-M pyronin Y Chemical compound [Cl-].C1=CC(=[N+](C)C)C=C2OC3=CC(N(C)C)=CC=C3C=C21 INCIMLINXXICKS-UHFFFAOYSA-M 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- AZJPTIGZZTZIDR-UHFFFAOYSA-L rose bengal Chemical compound [K+].[K+].[O-]C(=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 AZJPTIGZZTZIDR-UHFFFAOYSA-L 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229940101011 sodium hydroxymethylglycinate Drugs 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- YCUVUDODLRLVIC-VPHDGDOJSA-N sudan black b Chemical compound C1=CC(=C23)NC(C)(C)NC2=CC=CC3=C1\N=N\C(C1=CC=CC=C11)=CC=C1\N=N\C1=CC=CC=C1 YCUVUDODLRLVIC-VPHDGDOJSA-N 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 230000010415 tropism Effects 0.000 description 1
- 210000005239 tubule Anatomy 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- XOSXWYQMOYSSKB-LDKJGXKFSA-L water blue Chemical compound CC1=CC(/C(\C(C=C2)=CC=C2NC(C=C2)=CC=C2S([O-])(=O)=O)=C(\C=C2)/C=C/C\2=N\C(C=C2)=CC=C2S([O-])(=O)=O)=CC(S(O)(=O)=O)=C1N.[Na+].[Na+] XOSXWYQMOYSSKB-LDKJGXKFSA-L 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
-
- 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
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1429—Signal processing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1429—Signal processing
- G01N15/1433—Signal processing using image recognition
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
- G01N15/1436—Optical arrangements the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
- G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
- G01N15/147—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle the analysis being performed on a sample stream
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
- G01N21/53—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
- G01N33/4915—Blood using flow cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5094—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for blood cell populations
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B7/00—Mountings, adjusting means, or light-tight connections, for optical elements
- G02B7/28—Systems for automatic generation of focusing signals
- G02B7/36—Systems for automatic generation of focusing signals using image sharpness techniques, e.g. image processing techniques for generating autofocus signals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1019—Associating Coulter-counter and optical flow cytometer [OFC]
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
- G01N15/1409—Handling samples, e.g. injecting samples
- G01N2015/1411—Features of sheath fluids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
- G01N2015/1413—Hydrodynamic focussing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
- G01N2015/144—Imaging characterised by its optical setup
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
- G01N2015/1452—Adjustment of focus; Alignment
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1481—Optical analysis of particles within droplets
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N2015/1486—Counting the particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N21/05—Flow-through cuvettes
- G01N2021/058—Flat flow cell
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/24—Base structure
- G02B21/241—Devices for focusing
- G02B21/244—Devices for focusing using image analysis techniques
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B7/00—Mountings, adjusting means, or light-tight connections, for optical elements
- G02B7/28—Systems for automatic generation of focusing signals
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T2207/00—Indexing scheme for image analysis or image enhancement
- G06T2207/10—Image acquisition modality
- G06T2207/10141—Special mode during image acquisition
- G06T2207/10148—Varying focus
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T7/00—Image analysis
- G06T7/0002—Inspection of images, e.g. flaw detection
- G06T7/0012—Biomedical image inspection
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Dispersion Chemistry (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Ecology (AREA)
- Cell Biology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Signal Processing (AREA)
- Tropical Medicine & Parasitology (AREA)
- Computer Vision & Pattern Recognition (AREA)
- Physiology (AREA)
- Optics & Photonics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Optical Measuring Cells (AREA)
Abstract
本公开涉及一种染色方法,所述染色方法利用能够在单个步骤中快速将细胞染色的颗粒造影剂组合物。所述颗粒造影剂组合物可由一种或多种颗粒造影剂的组合、一种或多种渗透剂以及一种或多种固定剂构成。所述颗粒造影剂组合物可包括结晶紫、新亚甲蓝、皂草苷和戊二醛。
Description
相关专利申请的交叉引用
本专利申请要求2013年3月15日提交的、名称为“Analysis of Particles inFluid Samples(流体样品中的颗粒分析)”的美国专利申请第61/799,152号的权益,所述专利申请的全文以引用方式并入本文中。
技术领域
本公开总体涉及颗粒造影剂,更具体地,涉及颗粒造影剂组合物,所述颗粒造影剂组合物用于完全自动或部分自动装置中以鉴别和量化颗粒,如样品中的血细胞。
背景技术
血细胞分析是用于提供患者大致健康状况最常进行的医学检验之一。血液样品可从患者的身体抽取并保存在含有抗凝血剂以防止凝结的试管中。全血样品通常包括血细胞的三个主要类别,包括红血细胞(红细胞)、白血细胞(白细胞)和血小板(凝血细胞)。每一类可进一步分为具有多个成员的子类。例如,白血细胞(WBC)的五个主要类型或子类具有不同的形状和功能。白血细胞可包括嗜中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞和嗜碱性粒细胞。也有红血细胞类型的子类。样品中的颗粒外观可根据病理状况、细胞成熟度和其它原因而不同。红血细胞子类可包括网织红细胞和有核红血细胞。
估算RBC、WBC或血小板浓度的血细胞计数可手动进行或使用自动分析仪进行。当血细胞计数手动完成时,将一滴血施加到显微镜载片作为薄涂片。传统上,一直使用显微镜载片上的经干燥、染色的血液涂片的手工检查来确定五种白血细胞的数目或相对量。已使用组织学染料和染色剂(stain)来将细胞或细胞结构染色。例如,瑞氏染色剂(Wright'sstain)是已用来将在光学显微镜下检查的血涂片染色的组织学染色剂。将样品染色包括以适当的顺序使用多种溶液和步骤以确保染色剂正确施用,并且细胞结构被适当地保留。在第一步骤中,可将固定剂(fixing agent)施用于样品以保护样品不降解并保持细胞结构。随后,在第二步骤中,可将渗透剂施用于样品来溶解细胞膜,以允许染色剂进入细胞。在第三步骤中,可将染色剂施用于样品以将适当的结构染色。可将样品进一步漂洗用于观察,或可进行另外的步骤来施用另外的染色剂、复染色剂,或执行其它操作。
以适当的顺序持续适量的时间执行所述步骤是重要的。如果样品在被固色之前被渗透,则样品中的细胞结构可在被固色之前被降解,而且辨别原始细胞形态的任何能力都丧失。此外,染色不能先于渗透步骤发生,否则染色剂将无法正常穿透细胞并使细胞内的结构染色。此外,如果任何一个步骤,如固色、渗透和染色执行过快,则细胞形态可能丧失和/或细胞和其内部结构可能无法被正常染色。现有的染色技术需要多个步骤和大量时间。
现有的染色技术需要样品以通常约1:500或1:5000稀释于造影剂中。因此,现有的染色技术下正常的染色导致
自动分析仪正在变得越来越普及。全血细胞计数(CBC)可使用自动分析仪获得,其中的一种类型基于当颗粒或细胞沿小管通过感测区域时的阻抗或动态光散射来对血液样品中的不同颗粒或细胞计数。自动CBC可采用仪器或方法来区分不同类型的细胞(包括RBC、WBC和血小板(PLT)),所述不同类型的细胞可被独立地计数。例如,需要最小粒度或体积的计数技术可用来只对大细胞计数。血液中的某些细胞(如异常细胞)可能未被计数或正确识别。彼此附着的小细胞可能被错误地作为一个大细胞而计数。当怀疑错误计数时,可能需要手动复查仪器结果以验证和识别细胞。
自动血细胞计数技术可涉及流式细胞术。流式细胞术涉及提供狭窄流道,并对各个血细胞的通过进行感测和计数。流式细胞术方法已被用于检测悬浮在流体中的颗粒,如血液样品中的细胞,以及用于分析颗粒的类型、尺寸和体积分布,以推断出血液样品中相应颗粒类型的浓度或颗粒体积。用于分析悬浮于流体中的颗粒的合适方法的例子包括沉淀、微观表征、基于阻抗计数以及动态光散射。这些工具都会有测试误差。另一方面,颗粒的类型和浓度的准确表征在例如医疗诊断的应用中可能至关重要。
在基于成像的计数技术中,使用连接到数字照相机的显微镜物镜镜头捕捉可能正通过查看区域的制备样品的像素数据图像。像素图像数据可使用数据处理技术进行分析,并且还可显示在监视器上。
利用流动池(flowcell)的自动诊断系统的方面在Turner等人的美国专利第6,825,926号和Kasdan等人的美国专利第6,184,978号、第6,424,415号以及第6,590,646号中公开,这些专利均以引用方式并入,犹如在本文中充分阐述一样。
采用动态光散射或阻抗的自动系统已被用于获得全血细胞计数(CBC):总的白血细胞计数(WBC)、总的红血细胞的细胞体积(RBC分布)、血红蛋白HGB(血液中血红蛋白的量);平均细胞体积(MCV)(红细胞的平均体积);MPV(平均PLT体积);血细胞比容(HCT);MCH(HGB/RBC)(每个红血细胞的血红蛋白的平均量);以及MCHC(HGB/HCT)(细胞中血红蛋白的平均浓度)。已使用自动或部分自动工艺以有利于白血细胞五部分分类计数和血液样品分析。
以上所述的各种自动系统依赖于样品的快速分析。染色过程的步骤数和持续时间可能是自动颗粒分析系统的速度和功效的限制因素。如果染色过程缩短,则自动颗粒分析系统可能更有效率,并且如果染色过程以单一步骤进行,则自动颗粒分析系统会进一步更有效率。另外,如果样品的总大小被保持在最小,则自动颗粒分析系统可能更有效率。
发明内容
本发明公开了一种用于对在自动颗粒分析系统中成像的血液流体样品染色的颗粒造影剂组合物。所述颗粒造影剂组合物可包括选自结晶紫、新亚甲蓝、甲基绿、伊红Y以及番红O的至少一种颗粒造影剂。所述颗粒造影剂组合物可进一步包括选自表面活性剂、皂草苷、季铵盐、非离子表面活性剂、洗涤剂和两性离子表面活性剂的渗透剂。所述颗粒造影剂组合物可进一步包括选自戊二醛和甲醛的固定剂。
在一个实施例中,渗透剂可为皂草苷,所述皂草苷存在的量为在染色条件下足以得到介于约50mg/L和约750mg/L之间的浓度。固定剂可为戊二醛,所述戊二醛存在的量为在染色条件下足以得到0.1%或低于0.1%的浓度。
在一个实施例中,所述至少一种颗粒造影剂可包括结晶紫、新亚甲蓝以及伊红-Y。在染色条件下结晶紫与新亚甲蓝的比率可介于约1:90至约1:110之间。伊红-Y可以在染色条件下足以得到约3μM至约300μM的浓度的量存在。
在一个实施例中,结晶紫可以在染色条件下足以得到约6μM至约10μM的浓度的量存在。新亚甲蓝可以在染色条件下足以得到约70μM至约2.4mM的浓度的量存在。伊红-Y可以在染色条件下足以得到约10μM至约50μM的浓度的量存在。
在一些实施例中,结晶紫为大约90%的纯度或更纯。新亚甲蓝可为大约70%的纯度或更纯。伊红-Y可为大约80%的纯度或更纯。
在一些实施例中,结晶紫以在染色条件下足以得到约7.8μM的浓度的量存在。新亚甲蓝以在染色条件下足以得到约735μM的浓度的量存在。伊红-Y可以在染色条件下足以得到约27μM的浓度的量存在。在一些实施例中,颗粒造影剂组合物可另外包括缓冲组分。
本发明公开了一种用于处理血液流体样品的颗粒的方法,所述颗粒将使用自动颗粒分析系统成像。所述方法可包括将血液流体样品与颗粒造影剂组合物组合以获得样品混合物,以及将所述样品混合物在约37℃和约60℃之间的温度下温育少于90秒。所述颗粒造影剂组合物包括选自结晶紫、新亚甲蓝、甲基绿、伊红Y以及番红O的至少一种颗粒造影剂;选自表面活性剂、皂草苷、季铵盐、非离子表面活性剂、洗涤剂和两性离子表面活性剂的渗透剂;以及选自戊二醛和甲醛的固定剂。
在一些实施例中,所述颗粒造影剂可包括结晶紫、新亚甲蓝,结晶紫和新亚甲蓝的量为在染色条件下足以得到结晶紫与新亚甲蓝之间介于约1:1至约1:500的比率。皂草苷可以在染色条件下足以得到介于约50mg/L与约750mg/L之间的浓度的量被包括。戊二醛可以在染色条件下足以得到0.1%或低于0.1%的浓度的量被包括。所述方法可包括将样品混合物温育少于60秒。
在一些实施例中,所述颗粒造影剂组合物可包括结晶紫,结晶紫存在的量在染色条件下足以得到约6μM至约10μM的浓度。新亚甲蓝可以在染色条件下足以得到约70μM至约2.4mM的浓度的量存在。伊红-Y可以在染色条件下足以得到约10μM至约50μM的浓度的量存在。血液流体样品可与颗粒造影剂组合物以血液流体样品与颗粒造影剂组合物的比率为约1:2至约1:10进行组合。
在一些实施例中,所述方法可包括加热样品混合物至46℃到约49℃之间历时40到50秒之间。
在一些实施例中,结晶紫可为大约90%的纯度或更纯。新亚甲蓝可为大约70%的纯度或更纯。伊红-Y可为大约80%的纯度或更纯。
在一些实施例中,颗粒造影剂可包括结晶紫,结晶紫存在的量在染色条件下足以得到约7.8μM的浓度;新亚甲蓝,新亚甲蓝存在的量在染色条件下足以得到约735μM的浓度;以及伊红-Y,伊红-Y存在的量在染色条件下足以得到约27μM的浓度。所述颗粒造影剂组合物可进一步包括缓冲组分。血液流体样品可与颗粒造影剂组合物以血液流体样品与颗粒造影剂组合物的比率为约1:3至约1:4进行组合。可加热样品混合物至约47℃历时约45秒。
当结合附图来考虑时,通过参考下文的具体实施方式,本发明实施例的上述特征和许多其它特征以及伴随的优点将会变得显而易见并得到进一步理解。
附图说明
本说明书参照了下列附图,其中在不同图中使用相似的参考标记旨在表示相同或相似的部件。
图1是根据一个实施例用于输送样品流体的流动池的示意图。
图2是根据一个实施例制备颗粒造影剂组合物的示意图。
图3是根据一个实施例的快速、一步染色过程的流程图。
图4是根据一个实施例根据快速、一步染色过程染色的选择的白血细胞的代表性示图。
图5是根据一个实施例来自经颗粒造影剂组合物染色的样品的选择的白血细胞(selected while blood cells)的代表性示图。
图6是根据早期的实例1的经染色的细胞的代表性示图。
图7是根据早期的实例2的经染色的细胞的代表性示图。
图8是根据早期的实例3的经染色的细胞的代表性示图。
图9是根据早期的实例4的经染色的细胞的代表性示图。
图10是根据早期的实例的经染色的细胞的代表性示图。
图11是根据早期的实例5的经染色的细胞的代表性示图。
图12是根据早期的实例6的经染色的细胞的代表性示图。
具体实施方式
本公开涉及令人惊奇和意料不到的用于在样品中快速产生视觉区别的颗粒造影剂组合物。所述颗粒造影剂组合物尤其可用于自动流式细胞术系统中。所述颗粒造影剂组合物由颗粒造影剂的组合、渗透剂和固定剂组成。在一个实施例中,颗粒造影剂组合物是结晶紫、新亚甲蓝、皂草苷和戊二醛的混合物。在一个令人惊奇地有效的实施例中,在染色条件下,结晶紫存在的量足以得到约7.8μM的浓度,新亚甲蓝存在的量足以得到约735μM的浓度,皂草苷存在的量足以得到介于约50mg/L与约750mg/L之间的浓度,所述组合物进一步包括伊红-Y,其存在的量足以得到约27μM的浓度,以及戊二醛存在的量足以得到0.1%的浓度或低于0.1%的浓度。
提供这些示例性实例以向读者介绍本文讨论的一般主题,并非旨在限制所公开的构思的范围。以下部分参照附图描述了各种附加特征和实例,其中类似的标记表示类似的元件,且使用方向性描述来描述图示实施例,但是,如同图示实施例一样,它们不应当被用来限制本公开。在本文图示中包括的元件可能是未按比例绘制的。
本发明的颗粒造影剂组合物,当施用于血液流体样品时,使得这个样品中的细胞的染色类似于用标准血涂片染色剂处理的血涂片的染色,并且尤其类似于采用瑞氏染色剂(Wright’s stain)的血涂片的染色。瑞氏染色剂是一种组织学染色剂,它有利于区别血液细胞类型(例如WBC)。它主要用来将在光学显微镜下检查的外周血涂片和骨髓抽出物染色。在细胞遗传学中,它用于将染色体染色以方便症状和疾病的诊断。有以下一些已知的相关染色剂,如经缓冲的瑞氏染色剂、瑞氏-吉姆萨(Wright-Giemsa)染色剂以及经缓冲的瑞氏-吉姆萨染色剂。因为瑞氏染色过程包括醇溶剂,所以这种染色程序对活细胞是破坏性的,并且不能得到基本上完整的细胞。产生更强烈颜色的迈-格林华(May-Grünwald)染色剂也需要更长的时间来进行。
本发明的多个方面和实施例是基于令人惊奇和意想不到的发现,即某些颗粒造影剂组合物(包括例如染色剂/染料组合物,和/或它们的组合),当用于执行自动的、基于成像的样品分析(如血液分析)时,具有意想不到的性能和功效。
血液学-颗粒分析系统
本文公开的组合物和方法可用于许多不同类型的血液学成像系统。尤其是,本文所述的组合物和方法可用于基于图像的样品分析,如流动池分析。该流动池分析的一个实例可包括传统、已知的流式细胞术的方法。另外,本文所述的组合物和方法可有利地用于在以下简要描述的流动池分析系统和方法中,并进一步在2014年3月17日提交的名称为“Flowcell Systems And Methods For Particle Analysis In Blood Samples(用于血液样品中颗粒分析的流动池系统和方法)”、申请号为__/___,___以及2014年3月17日提交的名称为“Hematology Systems and Methods(血液学系统和方法)”、PCT申请号为________的共同提交的专利申请中进行描述,所述两个专利申请均以引用的方式并入本文中。
图1是用于将样品流体(例如以下所述的样品混合物)输送通过高光学分辨率成像装置24的观察区23的示例性流动池22的示意图,所述高光学分辨率成像装置24构造为使用数字图像处理使样品流(sample flow stream)32中的微观颗粒成像。流动池22连接到可能已经受处理(如与以下进一步详细描述的颗粒造影剂组合物接触)的样品流体源25。流动池22还连接到颗粒和/或细胞内细胞器校准液(PIOAL)的一个或多个源27,如粘度大于样品流体粘度的澄清甘油溶液。
样品流体经由样品进料管29的远端28的扁平开口注入,并且在PIOAL流已经基本上建立导致在带形样品流的上方和下方(或在相对侧上)的稳定和对称的PIOAL层流的点处进入流动池22内部。样品和PIOAL流可经由精密计量泵来提供,该精密计量泵使PIOAL与所注入的样品流体一起沿显著变窄的流动路径移动。PIOAL包裹并将样品流体压入流动路径变窄的区21中。因此,在区21中流动路径厚度的减小可有助于样品流32的几何集中。样品流体带32被包裹并在变窄区21的下游与PIOAL顺流而下,在高光学分辨率成像装置24的前方通过,或者说通过高光学分辨率成像装置24的查看区23,在该处例如使用CCD收集图像。样品流体带与PIOAL一起流至排放口33。
如此处所示,变窄区21可具有近侧流动路径部分21a(它具有近侧厚度PT)和远侧流动路径部分21b(它具有远侧厚度DT),使得远侧厚度DT小于近侧厚度PT。因此,可将样品流体在相对于近侧部分21a为远侧而相对于远侧部分21b为近侧的位置处通过样品管29的远端28注入。因此,当PIOAL被区21压缩时样品流体可进入PIOAL包层,其中样品流体注入管具有远侧出口,样品流体通过所述远侧出口被注入流动的护套流体(sheath fluid)中,所述远侧出口受流动池的流动路径尺寸的减少限制。
具有物镜镜头46的数字高光学分辨率成像装置24沿着与带形样品流32相交的光轴定向。物镜46和流动池33之间的相对距离经由操作电机驱动54可改变,以用于在光传感器阵列上分辨和收集聚焦的数字图像。
颗粒造影剂组合物
图2是根据一个实施例制备颗粒造影剂组合物的示意图。在方框208,将颗粒造影剂202、渗透剂204以及固定剂206组合生成颗粒造影剂组合物210。在一个实施例中,将颗粒造影剂202、渗透剂204以及固定剂206同时组合。在其它实施例中,以任何顺序将颗粒造影剂202、渗透剂204以及固定剂206中的一者与颗粒造影剂202、渗透剂204以及固定剂206中的另一者组合,随后与颗粒造影剂202、渗透剂204以及固定剂206中的最后一者组合。在方框208处的组合可以任何顺序并且以任何合适的方式进行。
在替代实施例中,渗透剂204和固定剂206中的一者不包括在颗粒造影剂组合物210中。在另外的实施例中,在方框208处其它物质被组合作为颗粒造影剂组合物210的一部分,如下文中进一步详细描述。
颗粒造影剂组合物210可作为试剂盒的一部分提供。颗粒造影剂组合物210可作为已制备好的形式提供,或作为必须要组合的一种或多种组分提供。
颗粒造影剂
颗粒造影剂202可为能够产生可见区别的任何造影剂,如那些类似于瑞氏染色剂的造影剂。此类造影剂的实例包括阿尔新蓝和阿尔新蓝86(PAS中性和酸性粘液物质);茜素红S;诱惑红AC(偶氮染料红色染料#40);苯胺蓝(用草酸强化的纤毛);金胺O;天蓝B;天蓝C;俾斯麦棕(Bismarck Brown);亮蓝FCF(考马斯蓝);亮甲酚蓝;亮绿;胭脂红(Carmium)(由胭脂红酸和钾明矾构成的红核染料);刚果红;Chlorozol黑E(核黑、细胞色素灰、糖原粉红);醋酸甲酚紫;达罗红(Darrow red);伊红蓝色;赤藓红B(红色染料#3);乙基伊红;固绿FCF(绿色染料#3);碱性品红(Fuchin basic)-(核和鞭毛);荧光素-(汞溴红);吉姆萨-外周血涂片;哈里斯苏木精(Harris hematoxylin)-退化核染色剂;靛蓝胭脂红(蓝色染料#2);詹纳斯绿B(Janus Green B)(线粒体);哲纳尔氏染色剂(Jenner Stain)-(外周血涂片);浅绿SF淡黄;麦克尼尔(MacNeal)-(四色血液染色剂);孔雀石绿;甲基橙;马休黄(Martiusyellow);梅尔氏苏木精(Mayer’s Hematoxylin)-渐进核染色剂;甲基紫2B;六次甲基四胺银(Methenamine Silver)-过碘酸;亚甲基紫;迈-格林华(May Grunwald)-血液染色剂;MTT-甲瓒染色剂(formazan stain);粘蛋白胭脂红-原发肿瘤染色剂;中性红;苯胺黑;尼罗蓝A;核固红C.I.60760;Napthal AS;硝基蓝四唑鎓-牢固甲瓒染料;橙色G;橙色II;地衣红;巴氏染色剂EAS(Papanicolaou Stain EAS)-鲜亮的细胞质染色;副品红(Pararosanilin);酸性品红(Pararosanaline);过碘酸雪夫氏(Periodic Acid Schiff)-(PAS,特异的碳水化合物染色剂);苯基喔星B(Phyloxine B);强蛋白银S(Protargol S);派洛宁B(Pyronin B);派洛宁Y;刃天青;罗曼诺斯基-吉姆萨(Romanowsky-Giemsa);孟加拉玫瑰红;番红O;苏丹黑B;苏丹III-(用α-萘酚染色骨髓颗粒);苏丹IV-染色三甘油酯;酒石黄-(偶氮染料黄色#5);硫堇(Thionin)-元核染色质染色剂;三苯基四唑鎓;TTC-甲瓒红色染料;甲苯胺蓝O;瑞氏染色剂-(用于常规血涂片的固定剂、缓冲剂和染色剂);以及瑞氏-吉姆萨。
通过不平凡的努力和试验,已经发现(如本文中进一步详细描述)使用包括结晶紫、新亚甲蓝、番红O、伊红Y以及甲基绿的至少一种颗粒造影剂202可获得具有令人惊奇地有效的结果的颗粒造影剂组合物210。以使活细胞和/或基本上完整的细胞染色的有效量添加颗粒造影剂202以便基于图像分类和子分类(subcategorization)。颗粒造影剂202可为上述颗粒造影剂的两种或更多种的任何组合。可选择颗粒造影剂202以有效地获得活细胞和/或基本上完整的细胞的“瑞氏样(Wright-like)”的染色图像。
在一个实施例中,颗粒造影剂202包括结晶紫。结晶紫可以在染色条件下足以达到介于约1μM至约100μM之间的量存在。如本文所用,术语“在染色条件下”是指当组分与样品混合时。结晶紫可以在染色条件下足以达到介于约6μM至约10μM之间的量存在。结晶紫可以在染色条件下足以达到约7.8μM的量存在。结晶紫可以在染色条件下足以达到非常接近7.8μM的量存在。结晶紫可被纯化至至少90%的纯度。结晶紫可被纯化至至少91%、92%、93%、94%、95%、96%、97%或98%的纯度。结晶紫可被纯化至至少99%的纯度。颗粒造影剂202可为单独结晶紫,或可为与一种或多种另外的颗粒造影剂组合的结晶紫。
在一个实施例中,颗粒造影剂202包括新亚甲蓝。新亚甲蓝可以在染色条件下足以达到介于约70μM至约2.4mM之间的量存在。新亚甲蓝可以在染色条件下足以达到介于约500μM至约950μM之间的量存在。新亚甲蓝可以在染色条件下足以达到约735μM的量存在。新亚甲蓝可以在染色条件下足以达到非常接近735μM的量存在。新亚甲蓝可被纯化至至少70%的纯度。新亚甲蓝可被纯化至至少71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的纯度。新亚甲蓝可被纯化至至少100%的纯度。
在一些实施例中,当颗粒造影剂202包括结晶紫和新亚甲蓝两者时获得令人惊奇地有效的结果。结晶紫与新亚甲蓝的比率可介于约1:1至约1:500(摩尔/摩尔)之间。结晶紫与新亚甲蓝的比率可介于约1:50至约1:160(摩尔/摩尔)之间。结晶紫与新亚甲蓝的比率可介于约1:90至约1:110(摩尔/摩尔)之间。
在一个实施例中,颗粒造影剂202包括伊红Y。伊红Y可以在染色条件下足以达到介于约3μM至约300μM之间的量存在。伊红Y可以在染色条件下足以达到介于约10μM至约50μM之间的量存在。伊红Y可以在染色条件下足以达到约27μM的量存在。伊红Y可以在染色条件下足以达到非常接近27μM的量存在。伊红Y可被纯化至至少80%的纯度。伊红Y可被纯化至至少81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的纯度。伊红Y可被纯化至至少100%的纯度。
在一些实施例中,当颗粒造影剂202是结晶紫、新亚甲蓝及伊红Y的组合,每一种具有如上所述的浓度和纯度的任何组合时,可获得令人惊奇地有效的结果。在一些实施例中,颗粒造影剂202具体来说是以足以达到约7.8μM的量存在的结晶紫、以足以达到约735μM的量存在的新亚甲蓝以及以足以达到约27μM的量存在的伊红Y。在一些实施例中,颗粒造影剂202具体来说是至少99%纯度的以足以达到约7.8μM的量存在的结晶紫、至少99%纯度的以足以达到约735μM的量存在的新亚甲蓝以及至少99%纯度的以足以达到约27μM的量存在的伊红Y。
在一个实施例中,颗粒造影剂202包括番红O。番红O可以在染色条件下足以达到介于约1μM至约100μM的量存在。番红O可以在染色条件下足以达到介于约3μM至约30μM的量存在。番红O可以在染色条件下足以达到约9μM的量存在。番红O可以在染色条件下足以达到非常接近9μM的量存在。番红O可被纯化至至少80%的纯度。番红O可被纯化至至少81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的纯度。番红O可被纯化至至少100%的纯度。
在一个实施例中,颗粒造影剂202包括甲基绿。甲基绿可以在染色条件下足以达到约0.1g/L的量存在。甲基绿可以在染色条件下足以达到非常接近0.1g/L的量存在。甲基绿可被纯化至至少80%的纯度。甲基绿可被纯化至至少81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%或99%的纯度。甲基绿可被纯化至至少100%的纯度。
在一些实施例中,颗粒造影剂202包括结晶紫、新亚甲蓝、番红O、伊红Y以及甲基绿中的一种或多种,它们存在的量使得当提供用于成像时能有效地产生对颗粒的视觉区别,例如经由增强样品中颗粒的细胞内内容物的特性产生区别。颗粒造影剂202可以足以使嗜中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞和嗜碱性粒细胞以及网织红细胞、有核红血细胞、血小板、母细胞、前髓细胞、髓细胞、后髓细胞或细胞碎片的亚细胞结构增强和/或染色。可视化或视觉区别可包括可利用任何光源(例如UV、可见光、IR)可视或者可检测的任何颗粒或颗粒内特性。
在颗粒造影剂组合物210包括两种或更多种颗粒造影剂202的实施例中,每种颗粒造影剂202的量可根据所述颗粒造影剂202对产生视觉区别以便进行颗粒分类和子分类是否具有独立、有竞争性和/或增强的影响来适当地调整。
渗透剂
在一些实施例中,渗透剂204可包括表面活性剂。在一些实施例中,渗透剂204可包括皂草苷。在替代实施例中,渗透剂204可包括季铵盐、非离子表面活性剂及两性离子表面活性剂的至少一种。渗透剂可改变细胞的渗透性,以增加颗粒造影剂202到细胞内内容物的可及性。渗透剂可经选择并以足以允许快速、一步染色过程的量被包括。
非离子表面活性剂的实例可包括(1)聚氧乙烯烷基醚或芳基醚(聚乙氧基化物),包括醚化为聚乙二醇或聚氧乙烯乙醇的直链脂肪族疏水物,例如35;(2)醚化为聚乙二醇的支链脂肪族/芳族(例如辛基酚)疏水物,例如Triton(3)醚化为聚乙二醇的直链脂肪族/芳族(例如正壬基苯酚)疏水物,例如C0897;以及(4)酯化为聚乙二醇的直链脂肪族(例如羧酸)疏水物,例如53,等等。非离子聚氧乙烯烷基或芳基醚(聚乙氧基化物)表面活性剂的实例可包括聚氧乙烯(4)月桂基醚(30);聚氧乙烯(23)月桂基醚(35);聚氧乙烯(2)鲸蜡基醚(52);聚氧乙烯(20)鲸蜡基醚(58);聚氧乙烯(2)硬脂基醚(72);聚氧乙烯(10)硬脂基醚(76);聚氧乙烯(20)硬脂基醚(78);聚氧乙烯(2)油基醚(92);聚氧乙烯(10)油基醚(96);聚氧乙烯(20)油基醚(98);聚氧乙烯(21)硬脂基醚(721);聚氧乙烯(100)硬脂基醚(700);等等。非离子表面活性剂的另外的实例可包括Triton(未还原的或还原的)、X-114(未还原的或还原的)、Triton以及Triton(未还原的或还原的),等等。
在一个实施例中,渗透剂204可包括35,其量在染色条件下足以得到约0.10g/L至约0.20g/L的浓度。35可以在染色条件下足以得到约0.10g/L至约0.16g/L的浓度的量存在。35可以足以得到约.012g/L至约0.14g/L的浓度的量存在。
两性离子表面活性剂的实例可包括TDAPS(四癸基二甲基铵基丙烷磺酸盐)、CHAPSO(3-[(3-胆酰胺丙基)二甲基铵基]-2-羟基-1-丙烷磺酸盐)、具有约12至约16个碳原子的烷基N,N-二甲基N-氧化物、月桂基二甲胺N-氧化物(LO)、DDAPS(正十二烷基-N,N-二甲基-3-铵基-1-丙烷磺酸盐),等等。
在一些实施例中,渗透剂204包括足以裂解红血细胞的试剂。在一些实施例中,渗透剂204包括足以裂解除网织红细胞或有核红血细胞之外的红血细胞的试剂。在一些实施例中,渗透剂204包括足以裂解红血细胞的试剂,而白血细胞、网织红细胞、有核红血细胞、血小板及其它细胞保持基本上完整。在一些实施例中,渗透剂204使白血细胞、网织红细胞、有核红血细胞和/或血小板的部分(member)和/或核膜可渗透性更强和/或更加多孔以有利于颗粒造影剂202进入。
在一些实施例中,渗透剂204选择成能够快速造成允许颗粒造影剂202进入样品中的细胞中的必需的孔或开口。
通过不平凡的努力和试验,已经发现,在使用包括得自佛罗里达劳德岱堡(Ft.Lauderdale,Florida)的临床诊断解决方案公司(CDS)的5PD-Lytic的渗透剂204的颗粒造影剂组合物210的一些实施例中可获得令人惊奇地有效的结果。5PD-Lytic包括皂草苷。5PD-Lytic在美国专利6,632,676中有一般性描述,该专利以引用的方式并入本文。
通过不平凡的努力和试验,已经发现,在使用包括皂草苷(所述皂草苷存在的量在染色条件下足以得到约10mg/L至约1000mg/L的浓度)的渗透剂204的颗粒造影剂组合物210的一些实施例中可获得令人惊奇地有效的结果。在一些实施例中,皂草苷以足以得到约50mg/L至约750mg/L的浓度的量存在。在一些实施例中,皂草苷可为季铵取代的皂草苷醚。
固定剂
在一些实施例中,固定剂206可选择成确保在染色和成像过程中白血细胞不降解。在一些实施例中,固定剂206可确保其它细胞和细胞结构不降解。固定剂的实例可包括戊二醛;甲醛;交联剂;等渗盐水中的苦味酸氨(例如用于亚甲蓝染色);乙醇;甲醇(例如在室温、-20℃或-70℃);海登海因氏苏萨液(Heidenhain’s Susa)-HgCl2、NaCl三氯乙酸、福尔马林;包因氏液(Bouin’s)-苦味酸、福尔马林、乙酸;杜波斯巴西液(Duboseq-Brazil)-具有80%EtOH的包因氏液;卡诺氏液(Carnoy’s)-EtOH、氯仿、乙酸;岑克尔氏液(Zenker’s)-HgCl2、K2CrO7、NaSO4.H2O;乙酸胭脂红;盖腾斯比液(Gatensby’s)-铬酸、四氧化锇、NaCl;贝克氏液(Baker’s)-福尔马林、CaCl2;史密斯氏液(Smith’s)-K2Cr2O7、福尔马林、乙酸;1%甲基绿、1%乙酸;苯酚、福尔马林、甘油、龙胆紫;萧丁液(Schaudin)-HgCl2、EtOH、乙酸;钱皮氏液(Champy’s)-铬酸、K2CrO7、OsO4;弗莱明氏液(Fleming’s)-铬酸(Cromic acid)、OsO4、乙酸;甲醛银液(Formol-Silver)-甲醛、AgNO3;施特雷克氏组织固定液(Streck’s TissueFixative)-溴代硝基丙二醇、二偶氮烷基脲、ZnSO4 .7H2O、柠檬酸钠;PBS中1%的咪唑烷基脲;乙二醛:Glyofix、Prefer、Safefix、Histochoice;1,3-二羟甲基-5,5-二甲基乙内酰脲-乙内酰脲;二羟甲基脲;N-羟甲基甘氨酸钠;卡诺夫斯基氏液(Karnovsky’s);水银氯(Mecuric chloride)(B-5);荷氏液(Hollande’s);等等。此外,合适的示例性固定剂可包括以下物质的任意一种或它们的任意组合。
在一些实施例中,固定剂206可为氧化剂、水银剂、苦味酸盐、羟乙基哌嗪乙磺酸-谷氨酸缓冲液介导的有机溶剂保护作用(HOPE)固定剂,或水溶性防腐剂。氧化剂的实例包括重铬酸钾、铬酸、高锰酸钾,等等。水银剂的实例包括B-5、岑克尔氏固定剂(Zernker’sfixative),等等。水溶性防腐剂的实例包括对羟基苯甲酸甲酯、羟基苯甲酸丙酯、二羟甲基脲、2-吡啶硫醇-1-氧化物、山梨酸、山梨酸钾,等等。
通过不平凡的努力和试验,已经发现使用包括戊二醛和甲醛的至少一种的固定剂206的颗粒造影剂组合物210的一些实施例中可获得令人惊奇地有效的结果。
在一些实施例中,使用包括0.1重量%或低于0.1重量%的戊二醛的固定剂206可获得令人惊奇地有效的结果。
其它组分
在一些实施例中,可选的其它组分212可在方框208可选地组合入颗粒造影剂组合物210中。其它组分212的实例可包括缓冲组分、粘度改良剂(modifying agent)、抗微生物剂、渗透调节剂、离子强度改性剂、表面活性剂、螯合剂,等等。在一些实施例中,当颗粒造影剂组合物210包括磷酸盐缓冲盐水时可获得令人惊奇地有效的结果。
示例性粘度改良剂包括天然水胶体(和衍生物),诸如角叉菜胶、刺槐豆胶、瓜尔豆胶和明胶;糖(和衍生物),诸如右旋糖、果糖;聚右旋糖;葡聚糖类;聚葡聚糖类;糖类;和多糖;半合成的水解胶体(和衍生物),诸如甲基纤维素、羧甲基纤维素;合成的水解胶体(和衍生物),诸如丙烯酸聚合物以及粘土(和衍生物),诸如膨润土和胶体硅酸镁铝
快速、一步染色过程
图3是根据一个实施例的快速、一步染色过程300的流程图。虽然快速、一步染色过程300可包含多个子步骤,术语“一步”用于标识该样品不需要在染色过程中引至多种不同的溶液中。颗粒造影剂组合物210在方框302制备,如上文参照图2所述。可选地,在一些实施例中,可在方框306纯化组分,如任意颗粒造影剂202。纯化颗粒造影剂202可降低与样品接触时形成的沉淀物的水平,从而减少背景并且改进基于图像的血液样品分析的结果,同时减少进一步复查图像或幻灯片或手动进行显微镜检查的需求。
在方框308,颗粒造影剂组合物210与样品组合。颗粒造影剂组合物210可以任何合适的方式与样品组合,包括混合在一起。方框308处的组合可包括用一定量的颗粒造影剂组合物210稀释样品。样品可用颗粒造影剂组合物210稀释。可选择稀释液的量以在基于图像的分析过程中每帧提供最佳数目的细胞。可选择稀释液的量以在基于图像的分析过程中每帧提供最佳数目的白血细胞。另外,可选择稀释液的量以为任何其它非基于图像的分析提供最佳体积。
通过不平凡的努力和试验,已经发现,在使用颗粒造影剂组合物210与样品的比率介于约2:1至约20:1的颗粒造影剂组合物210的一些实施例中可获得令人惊奇地有效的结果。颗粒造影剂组合物210与样品的比率可介于约3:1至约10:1之间。颗粒造影剂组合物210与样品的比率可介于约3:1至约4:1之间。颗粒造影剂组合物210与样品的比率可介于约3:1或约4:1。在一些实施例中,使用颗粒造影剂组合物210与样品的比率非常接近3:1或非常接近4:1可获得令人惊奇地有效的结果。
将颗粒造影剂与40mL 5PD-Lytic和50mL磷酸盐缓冲盐水一起使用,采用颗粒造影剂组合物210与样品的稀释比为10:1可获得令人惊奇地有效的结果。将颗粒造影剂与40mL5PD-Lytic、额外的皂草苷以及40mL磷酸盐缓冲盐水一起使用,采用颗粒造影剂组合物210与样品的稀释比为5:1可获得令人惊奇地有效的结果。将颗粒造影剂与40mL 5PD-Lytic、额外的皂草苷以及36mL磷酸盐缓冲盐水一起使用,采用颗粒造影剂组合物210与样品的稀释比为4:1可获得令人惊奇地有效的结果。
在一些实施例中,样品与颗粒造影剂组合物210在升高的温度(诸如下文结合温育所描述的任何温度)下组合。
如本文所用,组合的样品和颗粒造影剂组合物210称为样品混合物。
在方框310,将样品混合物在一定温度下温育一定量的时间。温育可增加细胞或它们的内部结构的渗透性,从而允许颗粒造影剂202更好地渗透到细胞或细胞结构中。可选择温育的时间和温度,以使颗粒造影剂组合物210能适当地渗透、固着和染色样品。可以选择温育的时间和温度,以确保红血细胞裂解,同时保持白血细胞、血小板和有核红血细胞基本上完好无损。
通过不平凡的努力和试验,已经发现当样品混合物在介于约37℃与约60℃之间的温度下温育约1至60秒时,在颗粒造影剂组合物210的一些实施例中可获得令人惊奇地有效的结果。可将样品混合物加热至介于约46℃约49℃之间的温度。样品混合物可温育40与50秒之间的时间。样品混合物可温育至多一小时。在一些实施例中,在约48℃下温育样品混合物约45秒可获得令人惊奇地有效的结果。在一些实施例中,在约47℃下温育样品混合物约45秒可获得令人惊奇地有效的结果。
在一些实施例中,方框308处的组合和方框310处的温育完成所需时间量与样品混合物在成像设备中处理和成像设备线被冲洗和/或清洁所需时间量大约相同或比之更短。以此方式,可使第一样品混合物成像的同时使第二样品混合物组合和温育。一旦第一样品混合物已成像并且成像设备已经被清洁,可立即使第二样品混合物成像。
在替代实施例中,方框308处的组合和方框310中处的温育完成所需时间量比样品混合物在成像设备中处理和成像设备线被冲洗和/或清洁所需时间量少不到两倍。以此方式,当第一样品混合物成像时,第二样品混合物可已准备好进行成像,并且第三样品混合物和第四样品混合物可处于混合及温育的过程中。一旦第一样品混合物已成像并且成像设备已经被清洁,可立即使第二样品混合物成像。第三样品混合物可正结束它的组合和温育,并且第四样品混合物可仍正进行组合和温育。一旦第二样品混合物已成像并且成像设备已经被清洁,可立即使第三样品混合物成像,同时第四样品混合物开始结束组合和温育,并且第五样品混合物开始进行组合和温育。该过程可无限继续以使样品混合物连续成像。
通过不平凡的努力和试验,已经发现,通过颗粒造影剂组合物210的某些实施例、组合颗粒造影剂组合物210与样品的某些方式以及温育样品混合物的某些方式的结合,可获得令人惊奇地有效的结果。
特别地,通过使用以下颗粒造影剂组合物210可获得令人惊奇地有效的结果:所述颗粒造影剂组合物210包括在染色条件下为约7.8μM的90%纯度或更纯的结晶紫、在染色条件下为约735μM的70%纯度或更纯的新亚甲蓝、在染色条件下为约27μM的80%纯度或更纯的伊红-Y、在染色条件下为约50mg/L至约750mg/L的经预处理的皂草苷,以及在染色条件下为约0.1%或0.1%以下的戊二醛;其中以介于约3:1与约4:1之间的颗粒造影剂210与样品的比率使颗粒造影剂210与样品组合;并且其中所得样品混合物在约48℃下温育约45秒。
某些有效的颗粒造影剂组合物210和染色程序使得活细胞和/或基本上完整的细胞的“瑞氏样”染色图像能够使用基于非醇的溶剂系统中的染料用自动视觉分析仪有效地获得。某些有效的颗粒造影剂组合物210和染色程序使得样品能够进行快速染色以使各种细胞组分、核叶以及颗粒结构可清楚地分辨。某些有效的颗粒造影剂组合物210和染色程序适用于离体活体染色。某些有效的颗粒造影剂组合物210和染色程序产生用于颗粒分类和子分类的视觉区别。某些有效的颗粒造影剂组合物210和染色程序可有效增强血清、脑脊髓液、胸膜液、滑液、精液、腹膜液、羊水、灌洗液、骨髓抽吸流体、积液、渗出液或血液样品中的细胞内颗粒内含物特征。某些有效的颗粒造影剂组合物210和染色程序可有效将嗜中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞、嗜碱性粒细胞、血小板、网织红细胞、有核红血细胞、母细胞、前髓细胞、髓细胞、后髓细胞、管型、细菌、上皮细胞和/或寄生生物染色。某些有效的颗粒造影剂组合物210和染色程序可有效产生视觉区别以用于颗粒分类和子分类,例如经由提供细胞的初级和次级颗粒的鉴别染色,基于该初级和次级颗粒的鉴别染色和增强(例如)来帮助未成熟粒细胞子分类以及确定它们的老化程度(age)。某些有效的颗粒造影剂组合物210和染色程序可有效产生视觉区别以用于计数或表征红血细胞、网织红细胞、有核红血细胞和血小板,以及用于白血细胞分类计数及白血细胞表征和分析。某些有效的颗粒造影剂组合物210和染色程序可有效地在活菌和/或活细胞和/或基本上保持完整结构的细胞中产生视觉区别。某些有效的颗粒造影剂组合物210和染色程序可有效地将嗜中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞、嗜碱性粒细胞,以及网织红细胞、有核红血细胞、血小板、母细胞、前髓细胞、髓细胞、后髓细胞或细胞碎片的的亚细胞结构染色。
经由本文所述的某些有效的颗粒造影剂组合物210和染色程序得以进行的快速染色可与手动或半自动成像和/或分析程序一起使用。
通过不平凡的努力和试验,已经发现,在颗粒造影剂组合物210的某些实施例中可获得令人惊奇地有效的结果,所述组合物210包含基于非醇的溶剂系统中的颗粒造影剂,所述造影剂能够(发明人第一次知道)产生活细胞和/或基本上完整的细胞的“瑞氏样”染色图像,这些图像可反映揭示各种细胞组分、核叶及粒状结构,并且使这些颗粒和/或细胞结构视觉上不同。
通过不平凡的努力和试验,已经发现,当使用由表1所列组分组成的颗粒造影剂组合物210时可获得令人惊奇地有效的结果,其中工作染色试剂(Working Stain Reagent)是由混合40mL新甲基蓝和5mL结晶紫来制得。
表1
50mL | 磷酸盐缓冲盐水 |
40mL | 工作染色试剂 |
40mL | 0.09%CDS 5PD-Lytic中的新甲基蓝 |
5mL | 0.009%CDS 5PD-Lytic中的结晶紫 |
10mL | 0.5%皂草苷 |
在染色条件下足以达到0.1%的量 | 戊二醛 |
图4是所选择的白血细胞的代表性示图,所述白血细胞来自用表1中所述的颗粒造影剂组合物210染色并且使用上文所述的快速、一步染色过程染色的样品。白血细胞是完整的且显示出瑞氏染色剂的染色特征。各种类型的白血细胞(例如嗜中性粒细胞、淋巴细胞、单核细胞、嗜酸性粒细胞、嗜碱性粒细胞等)在视觉上可分辨。
在一些实施例中,经本公开的颗粒造影剂组合物染色的细胞的特征记录在表2中。
表2
在某些实施例中,染色剂/染料组合物被配制成具有稳定性、便于储存、清理和/或有限的毒性。
图5是自以根据一个实施例的颗粒造影剂组合物210染色的样品选择的白血细胞的代表性示图,包括通过手动湿涂片成像和自动血流成像所成像的细胞。
早期实验
如参照以下实例所述,对许多染色组合物和方法进行了测试和修改,以产生以上公开的实施例。
在早期实例1中,存在两步染色方法,其中样品和颗粒造影剂组合物的早期实施例在47.5℃下组合并温育40秒,随后将骤冷试剂施用到样品混合物中。所述颗粒造影剂组合物包括库尔特LH系列稀释剂(Coulter LH Series Dilutent)、库尔特裂解S III差分裂解试剂(Coulter Lyse S III diff Lytic Reagent)、库尔特LH系列Pak试剂盒(Coulter LHSeries Pak Reagent Kit),以及库尔特LH系列网织红细胞PAK试剂盒(Coulter LH SeriesRETIC PAK Reagent Kit)。结果参见图6。
在早期实例1后的实例2中,实例1的两步染色法被一步染色法取代。相比于实例1的结果,嗜碱性粒细胞的改善的结果见图7。
在早期实例3中,不包括戊二醛的颗粒造影剂组合物导致白血细胞变弱,它们会由于流动池中的剪切力而碎裂。图8中示出显示实例3中损坏的膜的结果的图像。
在早期实例3后的实例4中,戊二醛被添加到颗粒造影剂组合物中。实例4中的白血细胞膜更完整,但核膜仍被损坏。在对PIOAL进行调整以减少甘油含量后,白血细胞的形态在成像过程中大多数没有变化,如图9中所示。
在使用新亚甲蓝和结晶紫的颗粒造影剂组合物的两染料染色剂的早期实例中,大多数细胞类型都很好分辨,除了嗜酸性粒细胞,它们一定程度上不一致并且不总是容易与嗜中性粒细胞区别开,如图10中所示。在后续的实例5和6中,将第三颗粒造影剂添加到颗粒造影剂组合物中。
在实例5中,将甲基绿添加到颗粒造影剂组合物中。甲基绿帮助使嗜酸性粒细胞染色更好,但所述细胞的核不再被染成想要的紫色,而成了蓝色。图11示出了实例5的具有蓝色染色的细胞核的嗜中性粒细胞的图像,但没有颗粒的细节。
在实例6中,将伊红-Y代替甲基绿用作颗粒造影剂组合物中的第三颗粒造影剂。伊红-y保留了核的紫红色染色,并且颗粒始终被略带橙色的光泽所染色,如图12中所见。
通过上述和另外的实验,已经确定,所公开的实施例和要求保护的实施例提供优选的结果。
本公开中所论述的所有专利、专利公布、专利申请、期刊论文、书籍、技术参考手册等全文以引用方式并入本文中以用于所有目的。
本文中所使用的任何标题仅用于组织目的,并且不被解释为以任何方式限制本公开或权利要求书。
在附图中描绘或上文所述的组件的不同布置,以及未示出或描述的组件和步骤也是可行的。相似地,一些特征结构和子组合是可用的,且它们可以在与其它特征结构和子组合无关的情况下被采用。出于例示性和非限制性的目的描述了本发明的实施例,但可供选择的实施例对于本专利的读者而言将是显而易见的。在某些情况下,方法步骤或操作可按不同的顺序执行或实施,或者可对操作进行添加、删除或修改。应当理解,在本发明的某些方面,可用多个部件来替换单个部件,并且可用单个部件来替换多个部件,以提供元件或结构或者执行给定的一种或多种功能。除了这种替换将不能有效实践本发明的某些实施例的情况之外,这种替换被视为在本发明的范围之内。因此,本发明不限于上文所述或在附图中示出的实施例,并且可在不脱离下面权利要求书的范围的前提下,做出各种实施例和进行各种修改。
Claims (11)
1.一种用于对在自动颗粒分析系统中成像的血液流体样品染色的颗粒造影剂组合物,包括:
结晶紫、新亚甲蓝和伊红- Y;
渗透剂,所述渗透剂包括皂草苷,所述皂草苷存在的量在染色条件下足以得到介于50mg/L与750mg/L之间的浓度;以及
固定剂,所述固定剂包括戊二醛,所述戊二醛存在的量在染色条件下足以得到0.1%或低于0.1%的浓度;
所述结晶紫与所述新亚甲蓝的比率在染色条件下介于1:90至1:110之间;并且
所述伊红-Y以在染色条件下足以得到3μM至300μM的浓度的量存在。
2.根据权利要求1所述的组合物,其中:
所述结晶紫以在染色条件下足以得到6μM至10μM的浓度的量存在;
所述新亚甲蓝以在染色条件下足以得到70μM至2.4mM的浓度的量存在;并且
所述伊红-Y以在染色条件下足以得到10μM至50μM的浓度的量存在。
3.根据权利要求2所述的组合物,其中:
所述结晶紫具有90%的纯度或更纯;
所述新亚甲蓝具有70%的纯度或更纯;并且
所述伊红-Y具有80%的纯度或更纯。
4.根据权利要求3所述的组合物,其中:
所述结晶紫以在染色条件下足以得到7.8μM的浓度的量存在;
所述新亚甲蓝以在染色条件下足以得到735μM的浓度的量存在;以及
所述伊红-Y以在染色条件下足以得到27μM的浓度的量存在。
5.根据权利要求2所述的组合物,还包括:
缓冲组分。
6.一种处理血液流体样品的颗粒的方法,所述血液流体样品将使用自动颗粒分析系统成像,所述方法包括:
将所述血液流体样品与颗粒造影剂组合物组合以获得样品混合物;以及
将所述样品混合物在介于37℃与60℃之间的温度下温育少于90秒;
其中所述颗粒造影剂组合物包括:
结晶紫、新亚甲蓝和伊红- Y;
渗透剂,所述渗透剂包括皂草苷,所述皂草苷存在的量在染色条件下足以得到介于50mg/L与750mg/L之间的浓度;以及
固定剂,所述固定剂包括戊二醛,所述戊二醛存在的量在染色条件下足以得到0.1%或低于0.1%的浓度;
所述结晶紫和所述新亚甲蓝的量在染色条件下足以得到所述结晶紫与所述新亚甲蓝之间介于1:1至1:500的比率;
所述伊红-Y存在的量在染色条件下足以得到10μM至50μM的浓度。
7.根据权利要求6所述的方法,其中:
所述温育所述样品混合物包括加热所述样品混合物少于60秒。
8.根据权利要求7所述的方法,其中:
所述颗粒造影剂组合物包括:
结晶紫,所述结晶紫存在的量在染色条件下足以得到6μM至10μM的浓度;
新亚甲蓝,所述新亚甲蓝存在的量在染色条件下足以得到70μM至2.4mM的浓度;以及
将所述血液流体样品与所述颗粒造影剂组合物的所述组合包括将所述血液流体样品与所述颗粒造影剂组合物以1:2至1:10的比率混合。
9.根据权利要求7所述的方法,其中温育所述样品混合物包括加热所述样品混合物至介于46℃与49℃之间历时介于40与50秒之间。
10.根据权利要求9所述的方法,其中:
所述结晶紫具有90%的纯度或更纯;
所述新亚甲蓝具有70%的纯度或更纯;以及
所述伊红-Y具有80%的纯度或更纯。
11.根据权利要求9所述的方法,其中:
所述颗粒造影剂组合物包括:
结晶紫,所述结晶紫存在的量在染色条件下足以得到7.8μM的浓度;
新亚甲蓝,所述新亚甲蓝存在的量在染色条件下足以得到735μM的浓度;
伊红-Y,所述伊红-Y存在的量在染色条件下足以得到27μM的浓度;以及
缓冲组分;
将所述血液流体样品与所述颗粒造影剂组合物的所述组合包括将所述血液流体样品与所述颗粒造影剂组合物以1:3至1:4的比率混合;以及
温育所述样品混合物包括加热所述样品混合物至47℃历时45秒。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201361799152P | 2013-03-15 | 2013-03-15 | |
US61/799,152 | 2013-03-15 | ||
PCT/US2014/030851 WO2014145984A1 (en) | 2013-03-15 | 2014-03-17 | Method and composition for staining and sample processing |
Publications (2)
Publication Number | Publication Date |
---|---|
CN105074422A CN105074422A (zh) | 2015-11-18 |
CN105074422B true CN105074422B (zh) | 2019-07-09 |
Family
ID=50631094
Family Applications (7)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810805317.0A Active CN109142195B (zh) | 2013-03-15 | 2014-03-17 | 用于体液样品中的粒子分析的自聚焦系统和方法 |
CN201480012645.5A Active CN105102959B (zh) | 2013-03-15 | 2014-03-17 | 用于血样中的颗粒分析的流通池系统和方法 |
CN201480013419.9A Active CN105074422B (zh) | 2013-03-15 | 2014-03-17 | 用于染色和样品处理的方法和组合物 |
CN201810941307.XA Pending CN109100288A (zh) | 2013-03-15 | 2014-03-17 | 用于血液样品中的粒子分析的鞘流体系统和方法 |
CN201480015291.XA Active CN105143850B (zh) | 2013-03-15 | 2014-03-17 | 用于血液样品中的粒子分析的自聚焦系统和方法 |
CN201480012697.2A Pending CN105164510A (zh) | 2013-03-15 | 2014-03-17 | 用于血液样品中的粒子分析的鞘流体系统和方法 |
CN201480015280.1A Active CN105143849B (zh) | 2013-03-15 | 2014-03-18 | 用于血液样品中粒子分析的动态范围扩展系统和方法 |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810805317.0A Active CN109142195B (zh) | 2013-03-15 | 2014-03-17 | 用于体液样品中的粒子分析的自聚焦系统和方法 |
CN201480012645.5A Active CN105102959B (zh) | 2013-03-15 | 2014-03-17 | 用于血样中的颗粒分析的流通池系统和方法 |
Family Applications After (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810941307.XA Pending CN109100288A (zh) | 2013-03-15 | 2014-03-17 | 用于血液样品中的粒子分析的鞘流体系统和方法 |
CN201480015291.XA Active CN105143850B (zh) | 2013-03-15 | 2014-03-17 | 用于血液样品中的粒子分析的自聚焦系统和方法 |
CN201480012697.2A Pending CN105164510A (zh) | 2013-03-15 | 2014-03-17 | 用于血液样品中的粒子分析的鞘流体系统和方法 |
CN201480015280.1A Active CN105143849B (zh) | 2013-03-15 | 2014-03-18 | 用于血液样品中粒子分析的动态范围扩展系统和方法 |
Country Status (10)
Country | Link |
---|---|
US (14) | US10705008B2 (zh) |
EP (8) | EP3489656B1 (zh) |
JP (4) | JP6404317B2 (zh) |
KR (4) | KR102055474B1 (zh) |
CN (7) | CN109142195B (zh) |
BR (4) | BR112015019969B1 (zh) |
ES (2) | ES2711364T3 (zh) |
HR (1) | HRP20181125T1 (zh) |
TR (2) | TR201809959T4 (zh) |
WO (4) | WO2014146051A1 (zh) |
Families Citing this family (91)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP4027130B1 (en) | 2007-02-01 | 2022-11-02 | Sysmex Corporation | Sample analyzer |
US9057676B2 (en) | 2010-02-05 | 2015-06-16 | Cytonome/St, Llc | Multiple flow channel particle analysis system |
KR101957923B1 (ko) | 2011-02-28 | 2019-03-14 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | 세포 배양 시스템 |
US9977188B2 (en) | 2011-08-30 | 2018-05-22 | Skorpios Technologies, Inc. | Integrated photonics mode expander |
CN105008159B (zh) | 2013-03-14 | 2017-10-10 | 伊利诺斯工具制品有限公司 | 压力释放阀 |
US9316635B2 (en) * | 2013-03-15 | 2016-04-19 | Iris International, Inc. | Sheath fluid systems and methods for particle analysis in blood samples |
CN109142195B (zh) | 2013-03-15 | 2021-10-01 | 艾瑞思国际股份有限公司 | 用于体液样品中的粒子分析的自聚焦系统和方法 |
US9857361B2 (en) | 2013-03-15 | 2018-01-02 | Iris International, Inc. | Flowcell, sheath fluid, and autofocus systems and methods for particle analysis in urine samples |
US20150030215A1 (en) * | 2013-07-26 | 2015-01-29 | Abbott Point Of Care, Inc. | Method and apparatus for producing an image of undiluted whole blood sample having wright stain coloration |
WO2015183992A1 (en) | 2014-05-27 | 2015-12-03 | Skorpios Technologies, Inc. | Waveguide mode expander using amorphous silicon |
EP3176563B1 (en) * | 2014-07-29 | 2021-06-30 | National University Corporation Hamamatsu University School of Medicine | Identification device and identification method |
AU2015307593B2 (en) * | 2014-08-28 | 2017-04-13 | Sysmex Corporation | Particle imaging apparatus and particle imaging method |
JP6616407B2 (ja) * | 2014-09-29 | 2019-12-04 | バイオサーフィット、 ソシエダッド アノニマ | フォーカシング方法 |
JP6704390B2 (ja) * | 2014-09-29 | 2020-06-03 | バイオサーフィット、 ソシエダッド アノニマ | 血球計数 |
WO2016050837A1 (en) * | 2014-09-30 | 2016-04-07 | Foss Analytical A/S | Method, device and system for hydrodynamic flow focusing |
BR112017007654B1 (pt) * | 2014-10-17 | 2020-12-01 | Iris International, Inc | método para imageamento de uma pluralidade de porções de fluido sanguíneo, e sistema para imageamento de uma pluralidade de porções de fluido sanguíneo |
JP6282969B2 (ja) * | 2014-10-31 | 2018-02-21 | 日本光電工業株式会社 | フロー解析装置、フローサイトメータ、及びフロー解析方法 |
JP6953678B2 (ja) * | 2014-11-12 | 2021-10-27 | 東洋紡株式会社 | 有形成分分析装置及び有形成分分析方法 |
JP6512593B2 (ja) * | 2015-02-23 | 2019-05-15 | 大日本印刷株式会社 | 培養液の培養状態解析システム及び培養状態解析方法、並びに、プログラム |
EP3957991A1 (en) * | 2015-03-31 | 2022-02-23 | Sysmex Corporation | Urine analysis system, image capturing apparatus, cell image capturing apparatus, urine analysis method, management apparatus, and information processing apparatus |
JP6713730B2 (ja) | 2015-05-20 | 2020-06-24 | シスメックス株式会社 | 細胞検出装置および細胞検出方法 |
CN105177978A (zh) * | 2015-07-10 | 2015-12-23 | 郎溪远华纺织有限公司 | 一种纺织布印染过程中的退浆方法 |
CN107850527A (zh) * | 2015-07-30 | 2018-03-27 | 皇家飞利浦有限公司 | 用于颗粒密度检测的激光传感器 |
CN105067488A (zh) * | 2015-08-11 | 2015-11-18 | 长春瑞克医疗科技有限公司 | 一种粒子成像室 |
WO2017057966A1 (ko) * | 2015-09-30 | 2017-04-06 | 주식회사 싸이토젠 | 세포 이미징 장치 및 그 방법 |
KR101873318B1 (ko) | 2015-09-30 | 2018-07-03 | 주식회사 싸이토젠 | 세포 이미징 장치 및 그 방법 |
EP3390963B1 (en) * | 2015-12-18 | 2021-02-17 | Abbott Laboratories | Method for determining the identity of histological stains |
WO2017108129A1 (de) | 2015-12-23 | 2017-06-29 | Siemens Healthcare Gmbh | Fliesszelle zur analyse von partikeln in einer zu untersuchenden flüssigkeit |
KR101807256B1 (ko) * | 2016-01-26 | 2017-12-08 | 한양대학교 에리카산학협력단 | 입자 분리 장치 및 입자 분리 방법 |
CN108780031A (zh) * | 2016-03-30 | 2018-11-09 | 西门子保健有限责任公司 | 使用环境粘弹性流体流对准样本流中的非球形生物实体 |
EP3440830A1 (en) | 2016-04-06 | 2019-02-13 | Biosurfit, S.A. | Method and system for capturing images of a liquid sample |
DE102016005974B4 (de) | 2016-05-13 | 2018-06-14 | Hochschule für Technik und Wirtschaft Dresden | Verfahren und Vorrichtung zur Einstellung des Laserfokus eines Anregungslasers in Blut durchflossenen Gefäßen für optische Messungen zur Bestimmung des Geschlechtes von Vogeleiern |
JP2017219479A (ja) * | 2016-06-09 | 2017-12-14 | 住友電気工業株式会社 | 微小粒子計測装置及び分析方法 |
CN109416313A (zh) * | 2016-06-24 | 2019-03-01 | 拜克门寇尔特公司 | 图像地图集系统和方法 |
EP3512935B1 (en) | 2016-09-13 | 2022-02-23 | President and Fellows of Harvard College | Methods relating to intestinal organ-on-a-chip |
CN109844494B (zh) * | 2016-10-06 | 2022-05-24 | 艾瑞斯国际有限公司 | 动态聚焦系统和方法 |
JP6783153B2 (ja) * | 2017-01-13 | 2020-11-11 | アークレイ株式会社 | フローセル及び測定装置 |
SG11201908847TA (en) | 2017-03-31 | 2019-10-30 | Life Technologies Corp | Apparatuses, systems and methods for imaging flow cytometry |
CN117054316A (zh) | 2017-05-19 | 2023-11-14 | 兴盛生物科技股份有限公司 | 用于计数细胞的系统和方法 |
CN107144521B (zh) * | 2017-06-06 | 2020-05-05 | 深圳小孚医疗科技有限公司 | 成像室姿态调节机构和调节方法 |
CN107144520B (zh) * | 2017-06-06 | 2020-05-05 | 深圳小孚医疗科技有限公司 | 用于显微成像粒子分析的粒子成像室和聚焦系统 |
CN107091800A (zh) * | 2017-06-06 | 2017-08-25 | 深圳小孚医疗科技有限公司 | 用于显微成像粒子分析的聚焦系统和聚焦方法 |
CN107643238A (zh) * | 2017-09-19 | 2018-01-30 | 中山大学附属第医院 | 一种定量检测血液循环微粒浓度的方法 |
US10466173B2 (en) * | 2017-10-06 | 2019-11-05 | Wyatt Technology Corporation | Optical flow cell assembly incorporating a replaceable transparent flow cell |
EP3697294B1 (en) * | 2017-10-16 | 2023-12-06 | Massachusetts Institute Of Technology | System and method for non-invasive hematological measurements |
US11609224B2 (en) | 2017-10-26 | 2023-03-21 | Essenlix Corporation | Devices and methods for white blood cell analyses |
CN108073547B (zh) * | 2017-12-06 | 2021-05-18 | 苏州大学 | 一种基于能量耗散的溶血经验预测方法及装置 |
US10274410B1 (en) * | 2018-01-23 | 2019-04-30 | Cbrn International, Ltd. | Bioaerosol particle detector |
CN108489872B (zh) * | 2018-03-23 | 2022-03-04 | 奥星制药设备(石家庄)有限公司 | 在线粒度监测方法和系统 |
CN108801740B (zh) * | 2018-03-28 | 2021-08-06 | 迈克生物股份有限公司 | 铁染色试剂 |
KR102026983B1 (ko) * | 2018-03-29 | 2019-09-30 | 두산중공업 주식회사 | 가스터빈 내 파이프를 통과하는 유체의 오염물을 모니터링 하는 시스템 및 그 방법 |
JP7109229B2 (ja) * | 2018-03-30 | 2022-07-29 | シスメックス株式会社 | フローサイトメーター及び粒子検出方法 |
SE542402C2 (en) * | 2018-07-02 | 2020-04-21 | Cellavision Ab | Method and apparatus for training a neural network classifier to classify an image depicting one or more objects of a biological sample |
EP3837543A4 (en) * | 2018-08-16 | 2022-11-16 | Essenlix Corporation | CELLULAR ANALYSIS IN BODY FLUIDS, ESPECIALLY BLOOD |
KR102100197B1 (ko) * | 2018-08-17 | 2020-04-14 | (주)엠큐빅 | 플로우 셀을 이용한 미세조류 연속 모니터링 장치 |
WO2020091720A1 (en) * | 2018-10-29 | 2020-05-07 | University Of Wyoming | Enhancement of fountain flow cytometry by reducing background light intensity |
US11802826B2 (en) * | 2018-10-29 | 2023-10-31 | Kyocera Corporation | Measurement apparatus |
CN109100286A (zh) * | 2018-10-31 | 2018-12-28 | 江苏卓微生物科技有限公司 | 细胞计数仪 |
WO2020133285A1 (zh) * | 2018-12-28 | 2020-07-02 | 深圳迈瑞生物医疗电子股份有限公司 | 一种血液细胞参数修正方法、血液样本检测仪和存储介质 |
CN109919946B (zh) * | 2019-02-19 | 2021-04-20 | 温州医科大学 | 一种基于光学相干层析成像技术预测巩膜透氧型接触镜镜后泪液形态变化止点的方法 |
CN113498362B (zh) * | 2019-02-27 | 2023-01-20 | 京瓷株式会社 | 粒子分离测量设备以及粒子分离测量装置 |
JP7274605B2 (ja) * | 2019-04-18 | 2023-05-16 | フンダシオ インスティテュート デ サイエンセズ フォトニクス | 個々の微生物の光学的調査のための流体装置 |
CN111760795B (zh) * | 2019-07-16 | 2022-02-01 | 北京京东乾石科技有限公司 | 用于分拣货物的方法和装置 |
CN116211296A (zh) | 2019-07-24 | 2023-06-06 | 麻省理工学院 | 用于甲襞成像装置的手指插入件 |
EP4042134A1 (en) | 2019-10-11 | 2022-08-17 | Beckman Coulter, Inc. | Method and composition for staining and sample processing |
US11125675B2 (en) * | 2019-10-18 | 2021-09-21 | Roger Lawrence Deran | Fluid suspended particle classifier |
WO2021118882A1 (en) * | 2019-12-10 | 2021-06-17 | Siemens Healthcare Diagnostics Inc. | Device for visualization of components in a blood sample |
US11877731B2 (en) * | 2020-03-07 | 2024-01-23 | Hall Labs Llc | Toilet with integral microscope slide |
CN111624065B (zh) * | 2020-05-22 | 2023-10-27 | 嘉兴优瑞生物科技有限公司 | 一种动物专用Diff-Quik染液及制备方法 |
US11860154B2 (en) | 2020-05-28 | 2024-01-02 | Leuko Labs, Inc | Method to detect white blood cells and/or white blood cell subtypes from non-invasive capillary videos |
CN111721950A (zh) * | 2020-05-29 | 2020-09-29 | 迪瑞医疗科技股份有限公司 | 一种稳定的有形成分分析聚焦液及其制备方法 |
JP2023550929A (ja) | 2020-11-19 | 2023-12-06 | ソニーグループ株式会社 | フレキシブルな画像ベースの粒子ソーティングのための分類ワークフロー |
CN112881246A (zh) * | 2021-01-26 | 2021-06-01 | 苏州胤煌精密仪器科技有限公司 | 一种可连续变倍的图像法粒度仪 |
US20240133779A1 (en) * | 2021-04-02 | 2024-04-25 | Noul Co., Ltd. | Hydrogel-based stamping for solution-free blood cell staining |
CN113326743B (zh) * | 2021-05-10 | 2023-10-13 | 大连海洋大学 | 一种养殖背景条件下鱼群运动行为参数提取和分析方法 |
WO2023010059A1 (en) * | 2021-07-27 | 2023-02-02 | Gateway Genomics, Llc | Methods, compositions, and kits for the preservation of nucleic acids |
US20230046207A1 (en) * | 2021-08-10 | 2023-02-16 | Becton, Dickinson And Company | Outlet fittings for reducing bubbles at the interface with a flow cell, and flow cytometers and methods using the same |
KR102572517B1 (ko) * | 2021-08-18 | 2023-08-31 | 주식회사 아폴론 | 인공지능 기반 호산구의 라만 데이터 처리 방법 및 장치 |
CN118159890A (zh) * | 2021-09-27 | 2024-06-07 | 贝克曼库尔特有限公司 | 可调节式安装设备 |
AU2022394321A1 (en) * | 2021-11-17 | 2024-03-21 | Becton, Dickinson And Company | Methods for dynamic real-time adjustment of a data acquisition parameter in a flow cytometer |
WO2023114204A1 (en) | 2021-12-17 | 2023-06-22 | Beckman Coulter, Inc. | Focus quality determination through multi-layer processing |
CN114441271B (zh) * | 2021-12-27 | 2023-06-27 | 桂林优利特医疗电子有限公司 | 一种新型染色液的配置及染色方法 |
WO2023129647A1 (en) | 2021-12-29 | 2023-07-06 | Beckman Coulter, Inc. | Biological sample driving system and method |
WO2023123466A1 (zh) * | 2021-12-31 | 2023-07-06 | 深圳迈瑞动物医疗科技股份有限公司 | 一种样本分析装置和样本分析方法 |
WO2023140235A1 (ja) * | 2022-01-20 | 2023-07-27 | 株式会社堀場製作所 | 粒子径分布測定装置、粒子径分布測定方法、及び粒子径分布測定用プログラム |
WO2023150064A1 (en) | 2022-02-02 | 2023-08-10 | Beckman Coulter, Inc. | Measure image quality of blood cell images |
WO2023172763A1 (en) | 2022-03-11 | 2023-09-14 | Beckman Coulter, Inc. | Controls and their use in analyzers |
WO2024020215A1 (en) | 2022-07-22 | 2024-01-25 | Beckman Coulter, Inc. | Biological sample staining module and biological analysis systems and methods |
FR3138211A1 (fr) * | 2022-07-25 | 2024-01-26 | Horiba Abx Sas | Dispositif pour le comptage et la différenciation de particules d’un flux d’échantillon |
WO2024030620A1 (en) * | 2022-08-05 | 2024-02-08 | Beckman Coulter, Inc. | Identification of immature cell types utilizing imaging |
WO2024123780A1 (en) | 2022-12-05 | 2024-06-13 | Beckman Coulter, Inc. | Hematology flow system |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1089659A (zh) * | 1993-01-14 | 1994-07-20 | 方奉谁 | 快速细胞染色剂 |
EP0656540A2 (en) * | 1993-10-08 | 1995-06-07 | Hitachi, Ltd. | Dyeing agent and apparatus for image analysis of flow type stain particles |
CN102822670A (zh) * | 2010-03-24 | 2012-12-12 | 贝克曼考尔特公司 | 用于分析血液样品的方法和系统 |
Family Cites Families (144)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3888782A (en) | 1972-05-08 | 1975-06-10 | Allergan Pharma | Soft contact lens preserving solution |
US3822095A (en) * | 1972-08-14 | 1974-07-02 | Block Engineering | System for differentiating particles |
US3819270A (en) * | 1972-10-02 | 1974-06-25 | Block Engineering | Blood cell analyzer |
GB1471976A (en) | 1974-09-20 | 1977-04-27 | Coulter Electronics | Particle sensing apparatus including a device for orienting generally flat particles |
DE2543310C2 (de) * | 1975-09-27 | 1982-04-29 | Gesellschaft für Strahlen- und Umweltforschung mbH, 8000 München | Einrichtung zum Zählen und Klassifizieren von in einer Flüssigkeit suspendierten Teilchen |
US4338024A (en) | 1980-05-02 | 1982-07-06 | International Remote Imaging Systems, Inc. | Flow analyzer and system for analysis of fluids with particles |
FR2484077B1 (fr) | 1980-06-06 | 1984-07-06 | Inst Nat Sante Rech Med | Procede et dispositif de mesure de la deformabilite de cellules vivantes, notamment des globules rouges du sang |
US4393466A (en) | 1980-09-12 | 1983-07-12 | International Remote Imaging Systems | Method of analyzing particles in a dilute fluid sample |
JPS5774372A (en) | 1980-10-27 | 1982-05-10 | Seiko Epson Corp | Fluid ink for printer |
DE3315195A1 (de) | 1982-04-29 | 1983-11-03 | International Remote Imaging Systems Inc., 91311 Chatsworth, Calif. | Verfahren zum ausrichten von teilchen in einer fluidprobe |
EP0125857B1 (en) * | 1983-05-12 | 1989-04-26 | Kabushiki Kaisha Toshiba | Grain counter |
US4647531A (en) * | 1984-02-06 | 1987-03-03 | Ortho Diagnostic Systems, Inc. | Generalized cytometry instrument and methods of use |
AU563260B2 (en) | 1984-11-29 | 1987-07-02 | International Remote Imaging Systems Inc. | Method of operating a microscopic instrument |
US4732479A (en) * | 1985-10-18 | 1988-03-22 | Canon Kabushiki Kaisha | Particle analyzing apparatus |
DE3851458T2 (de) | 1987-04-08 | 1995-02-09 | Hitachi Ltd | Vorrichtung mit einer scheideförmigen Durchflusszelle. |
JPS63262565A (ja) * | 1987-04-20 | 1988-10-28 | Hitachi Ltd | フロ−セル |
DE3902079A1 (de) | 1988-04-15 | 1989-10-26 | Bayer Ag | I.m. injektionsformen von gyrase-inhibitoren |
US5123055A (en) | 1989-08-10 | 1992-06-16 | International Remote Imaging Systems, Inc. | Method and an apparatus for differentiating a sample of biological cells |
JP2939647B2 (ja) * | 1990-07-24 | 1999-08-25 | シスメックス株式会社 | フローイメージングサイトメータにおける自動焦点調整方法 |
JP2874746B2 (ja) * | 1990-11-22 | 1999-03-24 | シスメックス株式会社 | フローイメージングサイトメータにおけるフローセル機構 |
JP3075370B2 (ja) * | 1991-07-26 | 2000-08-14 | シスメックス株式会社 | 粒子分析用のサンプル扁平流形成装置 |
US5412466A (en) * | 1991-07-26 | 1995-05-02 | Toa Medical Electronics Co., Ltd. | Apparatus for forming flattened sample flow for analyzing particles |
EP0556971B1 (en) | 1992-02-18 | 1999-12-08 | Hitachi, Ltd. | An apparatus for investigating particles in a fluid, and a method of operation thereof |
JP3111706B2 (ja) * | 1992-02-18 | 2000-11-27 | 株式会社日立製作所 | 粒子分析装置及び粒子分析方法 |
US5308526A (en) | 1992-07-07 | 1994-05-03 | The Procter & Gamble Company | Liquid personal cleanser with moisturizer |
JP3215175B2 (ja) | 1992-08-10 | 2001-10-02 | シスメックス株式会社 | 粒子分析装置 |
US5889881A (en) * | 1992-10-14 | 1999-03-30 | Oncometrics Imaging Corp. | Method and apparatus for automatically detecting malignancy-associated changes |
JP3052665B2 (ja) * | 1993-01-26 | 2000-06-19 | 株式会社日立製作所 | フローセル装置 |
US5585246A (en) | 1993-02-17 | 1996-12-17 | Biometric Imaging, Inc. | Method for preparing a sample in a scan capillary for immunofluorescent interrogation |
JP2826448B2 (ja) * | 1993-09-17 | 1998-11-18 | 株式会社日立製作所 | フロー式粒子画像解析方法およびフロー式粒子画像解析装置 |
JP3290786B2 (ja) | 1993-11-26 | 2002-06-10 | シスメックス株式会社 | 粒子分析装置 |
US5812419A (en) * | 1994-08-01 | 1998-09-22 | Abbott Laboratories | Fully automated analysis method with optical system for blood cell analyzer |
ES2278566T3 (es) | 1994-10-20 | 2007-08-16 | Sysmex Corporation | Reactivo y metodo para analizar componentes solidos en la orina. |
AU4741796A (en) | 1994-12-23 | 1996-07-19 | International Remote Imaging Systems Inc. | Method and apparatus of analyzing particles in a fluid sample and displaying same |
US5619032A (en) | 1995-01-18 | 1997-04-08 | International Remote Imaging Systems, Inc. | Method and apparatus for automatically selecting the best focal position from a plurality of focal positions for a focusing apparatus |
JPH0989753A (ja) * | 1995-09-25 | 1997-04-04 | Hitachi Ltd | 粒子分析装置 |
US5808737A (en) | 1996-02-29 | 1998-09-15 | Sienna Biotech, Inc. | Pre-analysis chamber for a flow particle analyzer |
WO1997043620A1 (en) | 1996-05-15 | 1997-11-20 | International Remote Imaging Systems, Inc. | Selectively emphasizing particles of interest from a fluid sample for analysis |
US6184978B1 (en) | 1996-05-15 | 2001-02-06 | International Remote Imaging Systems, Inc. | Method and apparatus for verifying uniform flow of a fluid sample through a flow cell and distribution on a slide |
US5872627A (en) * | 1996-07-30 | 1999-02-16 | Bayer Corporation | Method and apparatus for detecting scattered light in an analytical instrument |
KR100200734B1 (ko) | 1996-10-10 | 1999-06-15 | 윤종용 | 에어리얼 이미지 측정 장치 및 방법 |
US5985216A (en) | 1997-07-24 | 1999-11-16 | The United States Of America, As Represented By The Secretary Of Agriculture | Flow cytometry nozzle for high efficiency cell sorting |
US5935857A (en) * | 1997-08-01 | 1999-08-10 | Coulter International Corp. | Blood diluent |
US20020028471A1 (en) * | 1998-02-20 | 2002-03-07 | Oberhardt Bruce J. | Cell analysis methods and apparatus |
JP3867880B2 (ja) | 1998-04-08 | 2007-01-17 | シスメックス株式会社 | 尿中赤血球の鑑別装置および方法 |
DE69937353T2 (de) * | 1998-08-21 | 2008-07-17 | Union Biometrica, Inc., Somerville | Instrument zur analyse und selektiven verteilung von objektproben |
US20030059440A1 (en) | 1998-09-01 | 2003-03-27 | Tim Clarot | Composition and method for moisturizing nasal tissue |
US6130745A (en) * | 1999-01-07 | 2000-10-10 | Biometric Imaging, Inc. | Optical autofocus for use with microtiter plates |
JP2000214070A (ja) | 1999-01-21 | 2000-08-04 | Sysmex Corp | シ―スフロ―セルとそれを用いた血液分析装置 |
US6575930B1 (en) * | 1999-03-12 | 2003-06-10 | Medrad, Inc. | Agitation devices and dispensing systems incorporating such agitation devices |
JP4824157B2 (ja) | 1999-08-06 | 2011-11-30 | インベンテイオ・アクテイエンゲゼルシヤフト | 長いエスカレータと動く歩道の支持構造体 |
US6632676B1 (en) | 1999-09-24 | 2003-10-14 | Clinical Diagnostic Solutions | Multi-purpose reagent system and method for enumeration of red blood cells, white blood cells and thrombocytes and differential determination of white blood cells |
AU2730801A (en) | 1999-12-21 | 2001-07-03 | Yale University | Survivin promotion of angiogenesis |
JP2004500562A (ja) * | 1999-12-29 | 2004-01-08 | ユニオン バイオメトリカ インコーポレイテッド | フローサイトメトリーのための高粘度シース試薬 |
US6974938B1 (en) | 2000-03-08 | 2005-12-13 | Tibotec Bvba | Microscope having a stable autofocusing apparatus |
JP2001290161A (ja) | 2000-04-04 | 2001-10-19 | Advanced Display Inc | 液晶表示装置およびその製法 |
KR100347760B1 (ko) * | 2000-04-19 | 2002-08-09 | 엘지전자주식회사 | 기울어진 세탁조가 구비된 세탁기 |
CN1214340C (zh) | 2000-04-24 | 2005-08-10 | 国际遥距成象系统公司 | 多个神经网络的成像设备和方法 |
US7236623B2 (en) | 2000-04-24 | 2007-06-26 | International Remote Imaging Systems, Inc. | Analyte recognition for urinalysis diagnostic system |
US7283223B2 (en) * | 2002-08-21 | 2007-10-16 | Honeywell International Inc. | Cytometer having telecentric optics |
CA2349995A1 (en) | 2000-06-14 | 2001-12-14 | Lianne Ing | Viewing particles in a relatively translucent medium |
US7061595B2 (en) * | 2000-08-02 | 2006-06-13 | Honeywell International Inc. | Miniaturized flow controller with closed loop regulation |
US6875973B2 (en) * | 2000-08-25 | 2005-04-05 | Amnis Corporation | Auto focus for a flow imaging system |
AU2001290879A1 (en) * | 2000-09-15 | 2002-03-26 | California Institute Of Technology | Microfabricated crossflow devices and methods |
US20040070757A1 (en) * | 2000-12-29 | 2004-04-15 | Moore Richard Channing | High viscosity sheath reagent for flow cytometry |
FR2821428B1 (fr) * | 2001-02-23 | 2004-08-06 | Abx Sa | Reactif et procede pour l'identification et le comptage de cellules biologiques |
US7907765B2 (en) * | 2001-03-28 | 2011-03-15 | University Of Washington | Focal plane tracking for optical microtomography |
EP1395374B1 (en) * | 2001-05-17 | 2013-04-17 | Beckman Coulter, Inc. | Flow cytometer with active automated optical alignment system |
JP4838446B2 (ja) | 2001-06-20 | 2011-12-14 | オリンパス株式会社 | 顕微鏡装置 |
JP2003005088A (ja) | 2001-06-22 | 2003-01-08 | Nikon Corp | 顕微鏡用焦点合わせ装置およびそれを備えた顕微鏡 |
AU2002335715A1 (en) * | 2001-09-07 | 2003-03-24 | Burstein Technologies, Inc. | Optical bio-disc systems for nuclear morphology based identification |
JP4021183B2 (ja) | 2001-11-29 | 2007-12-12 | オリンパス株式会社 | 合焦状態信号出力装置 |
US20060050946A1 (en) * | 2002-05-10 | 2006-03-09 | Mitchison Timothy J | Computer-assisted cell analysis |
JP4370554B2 (ja) | 2002-06-14 | 2009-11-25 | 株式会社ニコン | オートフォーカス装置およびオートフォーカス付き顕微鏡 |
WO2004022774A1 (ja) * | 2002-09-05 | 2004-03-18 | Fuji Electric Systems Co.,Ltd. | 微生物または細胞の検出方法 |
US7319907B2 (en) | 2002-11-18 | 2008-01-15 | International Remote Imaging Systems, Inc. | Multi-level controller system |
US6825926B2 (en) | 2002-11-19 | 2004-11-30 | International Remote Imaging Systems, Inc. | Flow cell for urinalysis diagnostic system and method of making same |
JP2004188042A (ja) | 2002-12-13 | 2004-07-08 | Hitachi Hometec Ltd | 炊飯器 |
EP1447454A1 (en) * | 2003-02-14 | 2004-08-18 | DR. Chip Biotechnology Incorporation | Method and apparatus for detecting pathogens |
GB0304515D0 (en) * | 2003-02-27 | 2003-04-02 | Dakocytomation Denmark As | Standard |
WO2004077057A1 (en) * | 2003-02-27 | 2004-09-10 | Dakocytomation Denmark A/S | Standard for immunohistochemistry, immunocytochemistry and molecular cytogenetics |
DE10308171A1 (de) | 2003-02-27 | 2004-09-09 | Leica Microsystems Jena Gmbh | Verfahren zur automatischen Fokussierung |
US6900058B2 (en) * | 2003-03-11 | 2005-05-31 | Bionostics, Inc. | Control solution for photometric analysis |
US7324694B2 (en) | 2003-05-23 | 2008-01-29 | International Remote Imaging Systems, Inc. | Fluid sample analysis using class weights |
US20040241677A1 (en) | 2003-05-29 | 2004-12-02 | Lin Jeffrey S | Techniques for automated diagnosis of cell-borne anomalies with digital optical microscope |
JP4057539B2 (ja) * | 2004-01-09 | 2008-03-05 | 浜松ホトニクス株式会社 | シースフローセルキュベット及びその製造方法 |
US9176121B2 (en) | 2004-02-13 | 2015-11-03 | Roche Diagnostics Hematology, Inc. | Identification of blood elements using inverted microscopy |
US7394943B2 (en) | 2004-06-30 | 2008-07-01 | Applera Corporation | Methods, software, and apparatus for focusing an optical system using computer image analysis |
JP2006039315A (ja) | 2004-07-28 | 2006-02-09 | Hamamatsu Photonics Kk | 自動焦点装置及びそれを用いた顕微鏡装置 |
US7340957B2 (en) * | 2004-07-29 | 2008-03-11 | Los Alamos National Security, Llc | Ultrasonic analyte concentration and application in flow cytometry |
US7822276B2 (en) | 2005-02-17 | 2010-10-26 | Iris International, Inc. | Method and apparatus for analyzing body fluids |
CN101253401B (zh) | 2005-07-01 | 2013-01-02 | 霍尼韦尔国际公司 | 带三维流体动力学集中的模制标本盒 |
JP2007024844A (ja) * | 2005-07-21 | 2007-02-01 | Sysmex Corp | 血液分析方法及び血液分析装置 |
DE102005034441A1 (de) | 2005-07-22 | 2007-02-22 | Carl Zeiss Microimaging Gmbh | Mikroskopobjektiv |
EP1930717B1 (en) | 2005-09-29 | 2019-06-19 | Olympus Corporation | Focal position determining method and apparatus |
JP4896534B2 (ja) * | 2006-01-31 | 2012-03-14 | シスメックス株式会社 | 粒子分析装置用シース液 |
US20070209938A1 (en) * | 2006-03-13 | 2007-09-13 | Jianzhong Zhang | Method and apparatus for biopolymer analysis |
WO2007123744A2 (en) * | 2006-03-31 | 2007-11-01 | Solexa, Inc. | Systems and devices for sequence by synthesis analysis |
CN1834612A (zh) * | 2006-04-29 | 2006-09-20 | 北京索通医疗技术有限公司 | 一种用于血细胞分析仪的多功能稀释液及其制备方法 |
EP2487248A1 (en) * | 2006-05-10 | 2012-08-15 | The Board of Regents of the University of Texas System | Detecting tumor biomarker in oral cancer |
EP2028264A4 (en) | 2006-06-15 | 2012-10-24 | Nikon Corp | CELL INCUBATOR |
DE102006027836B4 (de) | 2006-06-16 | 2020-02-20 | Carl Zeiss Microscopy Gmbh | Mikroskop mit Autofokuseinrichtung |
SE530750C2 (sv) | 2006-07-19 | 2008-09-02 | Hemocue Ab | En mätapparat, en metod och ett datorprogram |
CN1945326A (zh) | 2006-10-13 | 2007-04-11 | 江西特康科技有限公司 | 基于视觉形态的五分类全血细胞分析方法 |
US7835000B2 (en) * | 2006-11-03 | 2010-11-16 | Los Alamos National Security, Llc | System and method for measuring particles in a sample stream of a flow cytometer or the like |
US7799575B2 (en) * | 2006-11-07 | 2010-09-21 | Genetix Limited | Flow cytometers |
JP2008276070A (ja) | 2007-05-02 | 2008-11-13 | Olympus Corp | 拡大撮像装置 |
WO2009014792A2 (en) | 2007-05-11 | 2009-01-29 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy | Electrical detection using confined fluids |
WO2008149365A2 (en) | 2007-06-07 | 2008-12-11 | Technion Research & Development Foundation Ltd. | Systems and methods for focusing particles |
US8941826B2 (en) * | 2007-09-10 | 2015-01-27 | The Penn State Research Foundation | Three-dimensional (3D) hydrodynamic focusing using a microfluidic device |
JP5393013B2 (ja) | 2007-10-04 | 2014-01-22 | 株式会社タイトー | 制御プログラム、Webサーバ及びゲーム配信システム |
JP2009089753A (ja) | 2007-10-04 | 2009-04-30 | Olympia:Kk | 遊技機、遊技機用プログラム、及び、遊技機用プログラムを記録したコンピュータ読み取り可能な記録媒体 |
US8266951B2 (en) * | 2007-12-19 | 2012-09-18 | Los Alamos National Security, Llc | Particle analysis in an acoustic cytometer |
US8714014B2 (en) * | 2008-01-16 | 2014-05-06 | Life Technologies Corporation | System and method for acoustic focusing hardware and implementations |
US9017610B2 (en) | 2008-04-25 | 2015-04-28 | Roche Diagnostics Hematology, Inc. | Method of determining a complete blood count and a white blood cell differential count |
US9602777B2 (en) | 2008-04-25 | 2017-03-21 | Roche Diagnostics Hematology, Inc. | Systems and methods for analyzing body fluids |
EP2290350B1 (en) * | 2008-06-04 | 2018-11-21 | Hitachi High-Technologies Corporation | Particle image analysis method and device |
CA2727478A1 (en) | 2008-06-13 | 2009-12-17 | Xy, Llc | Lubricious microfluidic flow path system |
RU2520848C2 (ru) * | 2008-06-30 | 2014-06-27 | Микробикс Байосистемз Инк. | Способ и прибор для сортировки клеток |
US8343978B2 (en) | 2008-08-04 | 2013-01-01 | Adds Pharmaceuticals Llc | Fast onset orodispersable tablets |
EP2331992A4 (en) | 2008-10-02 | 2014-12-17 | Pixcell Medical Technologies Ltd | VISCOELASTIC FOCUSING BASED OPTICAL IMAGING |
US20100221752A2 (en) * | 2008-10-06 | 2010-09-02 | Somalogic, Inc. | Ovarian Cancer Biomarkers and Uses Thereof |
FR2940976B1 (fr) | 2009-01-13 | 2017-11-03 | Oreal | Utilisation a des fins de criblage d'actifs anti-ages de formes solubles de la proteine de type desmogleine i |
JP5337912B2 (ja) | 2009-06-10 | 2013-11-06 | シンベニオ・バイオシステムズ・インコーポレーテッド | シース流装置及び方法 |
EP2288056A3 (en) * | 2009-07-22 | 2012-07-11 | Yamaha Corporation | Audio signal processing system comprising a plurality of devices connected by an audio network |
JP5586889B2 (ja) * | 2009-07-29 | 2014-09-10 | 株式会社日立ハイテクノロジーズ | 粒子画像解析装置 |
WO2011026136A1 (en) * | 2009-08-31 | 2011-03-03 | Life Technologies Corporation | Low-volume sequencing system and method of use |
US8362409B2 (en) | 2009-10-29 | 2013-01-29 | Applied Precision, Inc. | System and method for continuous, asynchronous autofocus of optical instruments |
US9068916B2 (en) * | 2010-03-15 | 2015-06-30 | Bio-Rad Laboratories, Inc. | Microassembled imaging flow cytometer |
CN102272574B (zh) * | 2010-03-31 | 2014-04-09 | 古河电气工业株式会社 | 光信息解析装置及光信息解析方法 |
US9090865B2 (en) * | 2010-10-29 | 2015-07-28 | The Regents Of The University Of California | Systems and methods for particle classification and sorting |
US8528427B2 (en) | 2010-10-29 | 2013-09-10 | Becton, Dickinson And Company | Dual feedback vacuum fluidics for a flow-type particle analyzer |
JP6121409B2 (ja) * | 2011-06-17 | 2017-04-26 | ロッシュ ダイアグノスティクス ヘマトロジー インコーポレイテッド | 生体試料の組織処理のための溶液 |
EP2568288A1 (en) | 2011-09-08 | 2013-03-13 | Koninklijke Philips Electronics N.V. | Means and methods for staining acid-fast cells |
KR101849974B1 (ko) | 2011-09-16 | 2018-04-19 | 삼성전자주식회사 | 개구수 제어 유닛, 이를 채용한 가변형 광 프로브 및 깊이 스캐닝 방법 |
US9810704B2 (en) | 2013-02-18 | 2017-11-07 | Theranos, Inc. | Systems and methods for multi-analysis |
US20140193892A1 (en) * | 2012-07-25 | 2014-07-10 | Theranos, Inc. | Image analysis and measurement of biological samples |
CA2920132C (en) * | 2012-10-24 | 2021-03-02 | The Regents Of The University Of California | System and method for deforming and analyzing particles |
CN102998437A (zh) | 2012-11-26 | 2013-03-27 | 江苏美诚生物科技有限公司 | 尿沉渣分析用鞘液及其制备方法 |
US9316635B2 (en) | 2013-03-15 | 2016-04-19 | Iris International, Inc. | Sheath fluid systems and methods for particle analysis in blood samples |
CN109142195B (zh) | 2013-03-15 | 2021-10-01 | 艾瑞思国际股份有限公司 | 用于体液样品中的粒子分析的自聚焦系统和方法 |
KR101772782B1 (ko) | 2013-03-15 | 2017-08-29 | 아이리스 인터내셔널 인크. | 소변 샘플을 염색 및 처리하기 위한 방법 및 조성물 |
US9857361B2 (en) | 2013-03-15 | 2018-01-02 | Iris International, Inc. | Flowcell, sheath fluid, and autofocus systems and methods for particle analysis in urine samples |
JP6112066B2 (ja) | 2014-05-27 | 2017-04-12 | 横河電機株式会社 | モジュール式電子機器連結用収納部材 |
-
2014
- 2014-03-17 CN CN201810805317.0A patent/CN109142195B/zh active Active
- 2014-03-17 JP JP2016502584A patent/JP6404317B2/ja active Active
- 2014-03-17 EP EP18213038.5A patent/EP3489656B1/en active Active
- 2014-03-17 JP JP2016502591A patent/JP6523246B2/ja active Active
- 2014-03-17 KR KR1020157024469A patent/KR102055474B1/ko active IP Right Grant
- 2014-03-17 US US14/216,811 patent/US10705008B2/en active Active
- 2014-03-17 BR BR112015019969-0A patent/BR112015019969B1/pt not_active IP Right Cessation
- 2014-03-17 EP EP21158285.3A patent/EP3842785B1/en active Active
- 2014-03-17 BR BR112015021593-9A patent/BR112015021593B1/pt not_active IP Right Cessation
- 2014-03-17 WO PCT/US2014/030928 patent/WO2014146051A1/en active Application Filing
- 2014-03-17 BR BR112015020098-2A patent/BR112015020098B1/pt not_active IP Right Cessation
- 2014-03-17 CN CN201480012645.5A patent/CN105102959B/zh active Active
- 2014-03-17 ES ES14722475T patent/ES2711364T3/es active Active
- 2014-03-17 EP EP14721152.8A patent/EP2972208B1/en active Active
- 2014-03-17 WO PCT/US2014/030902 patent/WO2014146030A1/en active Application Filing
- 2014-03-17 CN CN201480013419.9A patent/CN105074422B/zh active Active
- 2014-03-17 CN CN201810941307.XA patent/CN109100288A/zh active Pending
- 2014-03-17 TR TR2018/09959T patent/TR201809959T4/tr unknown
- 2014-03-17 US US14/217,034 patent/US10429292B2/en active Active
- 2014-03-17 JP JP2016502571A patent/JP6330025B2/ja active Active
- 2014-03-17 CN CN201480015291.XA patent/CN105143850B/zh active Active
- 2014-03-17 CN CN201480012697.2A patent/CN105164510A/zh active Pending
- 2014-03-17 WO PCT/US2014/030851 patent/WO2014145984A1/en active Application Filing
- 2014-03-17 ES ES14721152.8T patent/ES2683831T3/es active Active
- 2014-03-17 US US14/216,533 patent/US9322752B2/en active Active
- 2014-03-17 KR KR1020157024466A patent/KR102095617B1/ko active IP Right Grant
- 2014-03-17 EP EP14718878.3A patent/EP2972204B1/en active Active
- 2014-03-17 EP EP14722475.2A patent/EP2972211B1/en active Active
- 2014-03-17 KR KR1020157024768A patent/KR101802626B1/ko active IP Right Grant
- 2014-03-17 US US14/216,339 patent/US9279750B2/en active Active
- 2014-03-17 EP EP14723592.3A patent/EP2972214B1/en active Active
- 2014-03-17 TR TR2019/01203T patent/TR201901203T4/tr unknown
- 2014-03-17 EP EP18202980.1A patent/EP3467472B1/en active Active
- 2014-03-18 CN CN201480015280.1A patent/CN105143849B/zh active Active
- 2014-03-18 US US14/775,448 patent/US9702806B2/en active Active
- 2014-03-18 WO PCT/US2014/030939 patent/WO2014146061A1/en active Application Filing
- 2014-03-18 JP JP2016502593A patent/JP6401776B2/ja active Active
- 2014-03-18 KR KR1020157024439A patent/KR102067317B1/ko active IP Right Grant
- 2014-03-18 BR BR112015021800-8A patent/BR112015021800B1/pt not_active IP Right Cessation
- 2014-03-18 EP EP14722049.5A patent/EP2972210B1/en active Active
-
2016
- 2016-02-19 US US15/047,971 patent/US9909973B2/en active Active
-
2017
- 2017-07-10 US US15/645,710 patent/US10060846B2/en active Active
-
2018
- 2018-01-25 US US15/880,328 patent/US10345217B2/en active Active
- 2018-07-16 HR HRP20181125TT patent/HRP20181125T1/hr unknown
-
2019
- 2019-08-06 US US16/533,006 patent/US11525766B2/en active Active
- 2019-09-18 US US16/574,693 patent/US11543340B2/en active Active
-
2022
- 2022-11-09 US US17/984,116 patent/US11946848B2/en active Active
- 2022-11-09 US US17/984,125 patent/US20230075298A1/en active Pending
-
2023
- 2023-04-11 US US18/133,243 patent/US20230243730A1/en active Pending
- 2023-04-11 US US18/133,267 patent/US20230243731A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1089659A (zh) * | 1993-01-14 | 1994-07-20 | 方奉谁 | 快速细胞染色剂 |
EP0656540A2 (en) * | 1993-10-08 | 1995-06-07 | Hitachi, Ltd. | Dyeing agent and apparatus for image analysis of flow type stain particles |
CN102822670A (zh) * | 2010-03-24 | 2012-12-12 | 贝克曼考尔特公司 | 用于分析血液样品的方法和系统 |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105074422B (zh) | 用于染色和样品处理的方法和组合物 | |
CN105102960B (zh) | 用于对尿液样品进行染色和处理的方法和组合物 | |
US10774359B2 (en) | Cellular analysis of body fluids | |
EP0179107B1 (en) | Metachromatic dye sorption means for differential determination of sub-populations of lymphocytes | |
CN105074420A (zh) | 血液学系统和方法 | |
JPH06507971A (ja) | 絶対白血球サブセット計数値および白血球の多数の部の差を得るための方法および装置 | |
US20210108994A1 (en) | Method and composition for staining and sample processing | |
US20210033592A1 (en) | Blood analyzer and analysis method | |
CN103398935B (zh) | 一种用于白细胞分类计数的方法以及试剂盒 | |
CN106404638A (zh) | 基于多元技术的贝类血淋巴细胞分类方法 | |
DE60031521T2 (de) | Produkte und verfahren für einzelwert- und vielfachwert-phänotypisierung von zellen | |
JPH1183849A (ja) | 尿中有形成分の分析方法及びその分析試薬 | |
Kavati et al. | A simple, fast, inexpensive and efficient method for leukocytes separation with preservation of morphology and cell viability for use in education and research | |
Tvedten | Routine stains and automated stainers | |
US20230204580A1 (en) | Formulation for total and differential counting of leukocytes in liquid medium and method of making and using same | |
EP3392343A1 (en) | Water-based cell staining composition, method for preparing same and use thereof, cell sample preparation and cell counting methods |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |