JP2007071890A - 白血球の分類計数方法 - Google Patents
白血球の分類計数方法 Download PDFInfo
- Publication number
- JP2007071890A JP2007071890A JP2006319621A JP2006319621A JP2007071890A JP 2007071890 A JP2007071890 A JP 2007071890A JP 2006319621 A JP2006319621 A JP 2006319621A JP 2006319621 A JP2006319621 A JP 2006319621A JP 2007071890 A JP2007071890 A JP 2007071890A
- Authority
- JP
- Japan
- Prior art keywords
- leukocytes
- scattered light
- classifying
- intensity
- sample
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 29
- 239000003219 hemolytic agent Substances 0.000 claims abstract description 15
- 230000002489 hematologic effect Effects 0.000 claims abstract description 12
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 5
- 238000005259 measurement Methods 0.000 claims abstract description 5
- 210000000265 leukocyte Anatomy 0.000 claims description 104
- 239000000975 dye Substances 0.000 claims description 26
- 208000010110 spontaneous platelet aggregation Diseases 0.000 claims description 13
- 210000004369 blood Anatomy 0.000 claims description 10
- 239000008280 blood Substances 0.000 claims description 10
- 239000007850 fluorescent dye Substances 0.000 claims description 10
- 238000010186 staining Methods 0.000 claims description 4
- 230000002934 lysing effect Effects 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 230000002949 hemolytic effect Effects 0.000 claims 1
- 230000002776 aggregation Effects 0.000 abstract description 2
- 238000004220 aggregation Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 41
- -1 polyoxyethylene Polymers 0.000 description 22
- 230000035484 reaction time Effects 0.000 description 20
- 230000002159 abnormal effect Effects 0.000 description 19
- 125000000217 alkyl group Chemical group 0.000 description 19
- 239000000523 sample Substances 0.000 description 15
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 12
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 210000003714 granulocyte Anatomy 0.000 description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 7
- 210000000601 blood cell Anatomy 0.000 description 7
- 239000004065 semiconductor Substances 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
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 235000001014 amino acid Nutrition 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
- 210000000170 cell membrane Anatomy 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 210000004698 lymphocyte Anatomy 0.000 description 6
- 210000001616 monocyte Anatomy 0.000 description 6
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 5
- 210000001185 bone marrow Anatomy 0.000 description 5
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 5
- 238000000691 measurement method Methods 0.000 description 5
- 229930182817 methionine Natural products 0.000 description 5
- 235000006109 methionine Nutrition 0.000 description 5
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 229910052786 argon Inorganic materials 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 229910052736 halogen Inorganic materials 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000002736 nonionic surfactant Substances 0.000 description 4
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 4
- 108700004121 sarkosyl Proteins 0.000 description 4
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 239000007995 HEPES buffer Substances 0.000 description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- BACYUWVYYTXETD-UHFFFAOYSA-N N-Lauroylsarcosine Chemical compound CCCCCCCCCCCC(=O)N(C)CC(O)=O BACYUWVYYTXETD-UHFFFAOYSA-N 0.000 description 3
- 125000002252 acyl group Chemical group 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000004820 blood count Methods 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 125000004432 carbon atom Chemical group C* 0.000 description 3
- 125000002091 cationic group Chemical group 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 3
- 229960005542 ethidium bromide Drugs 0.000 description 3
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 3
- 150000002367 halogens Chemical class 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- 230000003204 osmotic effect Effects 0.000 description 3
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical class C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 229910052717 sulfur Inorganic materials 0.000 description 3
- CMCBDXRRFKYBDG-UHFFFAOYSA-N 1-dodecoxydodecane Chemical compound CCCCCCCCCCCCOCCCCCCCCCCCC CMCBDXRRFKYBDG-UHFFFAOYSA-N 0.000 description 2
- UMCMPZBLKLEWAF-BCTGSCMUSA-N 3-[(3-cholamidopropyl)dimethylammonio]propane-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)CCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 UMCMPZBLKLEWAF-BCTGSCMUSA-N 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- 0 CNI1=CC=CC=C1SC(C=**=C1C=CN(CC(CO)O)C2C=CC=C[C@]12)=I Chemical compound CNI1=CC=CC=C1SC(C=**=C1C=CN(CC(CO)O)C2C=CC=C[C@]12)=I 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 2
- 239000004470 DL Methionine Substances 0.000 description 2
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 2
- QTANTQQOYSUMLC-UHFFFAOYSA-O Ethidium cation Chemical compound C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 QTANTQQOYSUMLC-UHFFFAOYSA-O 0.000 description 2
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- 125000003342 alkenyl group Chemical group 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 125000000304 alkynyl group Chemical group 0.000 description 2
- 150000001450 anions Chemical class 0.000 description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 2
- 229910052794 bromium Inorganic materials 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 229910052801 chlorine Inorganic materials 0.000 description 2
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical class 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)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 2
- 239000002812 cholic acid derivative Substances 0.000 description 2
- 230000001332 colony forming effect Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 239000011737 fluorine Substances 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 125000001165 hydrophobic group Chemical group 0.000 description 2
- 239000011630 iodine Substances 0.000 description 2
- 229910052740 iodine Inorganic materials 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- FFEARJCKVFRZRR-UHFFFAOYSA-N methionine Chemical compound CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 2
- 210000003887 myelocyte Anatomy 0.000 description 2
- 125000001421 myristyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- GCRLIVCNZWDCDE-SJXGUFTOSA-N n-methyl-n-[(2s,3r,4r,5r)-2,3,4,5,6-pentahydroxyhexyl]nonanamide Chemical compound CCCCCCCCC(=O)N(C)C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO GCRLIVCNZWDCDE-SJXGUFTOSA-N 0.000 description 2
- SBWGZAXBCCNRTM-CTHBEMJXSA-N n-methyl-n-[(2s,3r,4r,5r)-2,3,4,5,6-pentahydroxyhexyl]octanamide Chemical compound CCCCCCCC(=O)N(C)C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO SBWGZAXBCCNRTM-CTHBEMJXSA-N 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 210000004765 promyelocyte Anatomy 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 125000004434 sulfur atom Chemical group 0.000 description 2
- 125000004191 (C1-C6) alkoxy group Chemical group 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
- UNWFFCPRJXMCNV-UHFFFAOYSA-N 3-[dodecanoyl(methyl)amino]propanoic acid Chemical compound CCCCCCCCCCCC(=O)N(C)CCC(O)=O UNWFFCPRJXMCNV-UHFFFAOYSA-N 0.000 description 1
- BGWLYQZDNFIFRX-UHFFFAOYSA-N 5-[3-[2-[3-(3,8-diamino-6-phenylphenanthridin-5-ium-5-yl)propylamino]ethylamino]propyl]-6-phenylphenanthridin-5-ium-3,8-diamine;dichloride Chemical compound [Cl-].[Cl-].C=1C(N)=CC=C(C2=CC=C(N)C=C2[N+]=2CCCNCCNCCC[N+]=3C4=CC(N)=CC=C4C4=CC=C(N)C=C4C=3C=3C=CC=CC=3)C=1C=2C1=CC=CC=C1 BGWLYQZDNFIFRX-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- GHUXAYLZEGLXDA-UHFFFAOYSA-N 8-azido-5-ethyl-6-phenylphenanthridin-5-ium-3-amine;bromide Chemical compound [Br-].C12=CC(N=[N+]=[N-])=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 GHUXAYLZEGLXDA-UHFFFAOYSA-N 0.000 description 1
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 1
- IVRMZWNICZWHMI-UHFFFAOYSA-N Azide Chemical compound [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 239000006173 Good's buffer Substances 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 208000028018 Lymphocytic leukaemia Diseases 0.000 description 1
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 1
- 108010077895 Sarcosine Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- DPXHITFUCHFTKR-UHFFFAOYSA-L To-Pro-1 Chemical compound [I-].[I-].S1C2=CC=CC=C2[N+](C)=C1C=C1C2=CC=CC=C2N(CCC[N+](C)(C)C)C=C1 DPXHITFUCHFTKR-UHFFFAOYSA-L 0.000 description 1
- QHNORJFCVHUPNH-UHFFFAOYSA-L To-Pro-3 Chemical compound [I-].[I-].S1C2=CC=CC=C2[N+](C)=C1C=CC=C1C2=CC=CC=C2N(CCC[N+](C)(C)C)C=C1 QHNORJFCVHUPNH-UHFFFAOYSA-L 0.000 description 1
- MZZINWWGSYUHGU-UHFFFAOYSA-J ToTo-1 Chemical compound [I-].[I-].[I-].[I-].C12=CC=CC=C2C(C=C2N(C3=CC=CC=C3S2)C)=CC=[N+]1CCC[N+](C)(C)CCC[N+](C)(C)CCC[N+](C1=CC=CC=C11)=CC=C1C=C1N(C)C2=CC=CC=C2S1 MZZINWWGSYUHGU-UHFFFAOYSA-J 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007933 aliphatic carboxylic acids Chemical class 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229910052796 boron Inorganic materials 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 230000000093 cytochemical effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 125000005066 dodecenyl group Chemical group C(=CCCCCCCCCCC)* 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 210000003617 erythrocyte membrane Anatomy 0.000 description 1
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- UQSQSQZYBQSBJZ-UHFFFAOYSA-N fluorosulfonic acid Chemical compound OS(F)(=O)=O UQSQSQZYBQSBJZ-UHFFFAOYSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 125000005843 halogen group Chemical group 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 208000003747 lymphoid leukemia Diseases 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- JZMJDSHXVKJFKW-UHFFFAOYSA-M methyl sulfate(1-) Chemical compound COS([O-])(=O)=O JZMJDSHXVKJFKW-UHFFFAOYSA-M 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 210000001167 myeloblast Anatomy 0.000 description 1
- 210000003643 myeloid progenitor cell Anatomy 0.000 description 1
- UMWKZHPREXJQGR-UHFFFAOYSA-N n-methyl-n-(2,3,4,5,6-pentahydroxyhexyl)decanamide Chemical compound CCCCCCCCCC(=O)N(C)CC(O)C(O)C(O)C(O)CO UMWKZHPREXJQGR-UHFFFAOYSA-N 0.000 description 1
- UMWKZHPREXJQGR-XOSAIJSUSA-N n-methyl-n-[(2s,3r,4r,5r)-2,3,4,5,6-pentahydroxyhexyl]decanamide Chemical compound CCCCCCCCCC(=O)N(C)C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO UMWKZHPREXJQGR-XOSAIJSUSA-N 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000001117 oleyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])/C([H])=C([H])\C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 125000001147 pentyl group Chemical group C(CCCC)* 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 125000001501 propionyl group Chemical group O=C([*])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229940043230 sarcosine Drugs 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- KSAVQLQVUXSOCR-UHFFFAOYSA-M sodium lauroyl sarcosinate Chemical compound [Na+].CCCCCCCCCCCC(=O)N(C)CC([O-])=O KSAVQLQVUXSOCR-UHFFFAOYSA-M 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 125000005063 tetradecenyl group Chemical group C(=CCCCCCCCCCCCC)* 0.000 description 1
- 125000002889 tridecyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000002948 undecyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Abstract
【解決手段】 白血球の分類計数方法は、(1)血液学的試料を、赤血球を溶解させるための溶血剤によって処理する工程、(2)処理された試料をフローサイトメータに導入して、少なくとも散乱光を測定する工程、(3)前方散乱光幅の差を利用して、血小板凝集と、白血球とを分類する工程からなる。
【選択図】 図1
Description
本発明は前記事情を鑑み、白血球を測定するための迅速、簡便かつ高精度に測定可能な白血球の分類計数方法を提供するものである。
(1)血液学的試料を、赤血球を溶解させるための溶血剤によって処理する工程、
(2)処理された試料をフローサイトメータに導入して、少なくとも散乱光を測定する工程、
(3)前方散乱光幅の差を利用して、血小板凝集と、白血球とを分類する工程
からなる白血球の分類計数方法である。
式(III):
MEGA8(オクタノイル−N−メチルグルカミド)、MEGA9(ノナイノイル−N−メチルグルカミド)、MEGA10(デカノイル−N−メチルグルカミド)等が好適に使用できる。
150〜600mOsm/kgにする緩衝液、電気伝導度を6.0〜
9.0mS/cmにするための緩衝液からなる、特開平6−273413号に記載の溶血剤が好ましい。
図18は、本実施形態に使用できるフローサイトメータの光学系を示す斜視図である。同図においてレーザ21から出射されたビームはコリメートレンズ22を介してシースフローセル23のオリフィス部を照射する。オリフィス部を通過する血球から発せられる前方散乱光は集光レンズ24とピンホール板25を介してフォトダイオード26に入射する。
ポリオキシエチレン(16)オレイルエーテル 24.0g
N−ラウロイルサルコシン酸ナトリウム 1.5g
DL−メチオニン 20.0g
1N−NaOH 0.3g
NaCl 4.0g
式(VI)の色素 3.0mg
HEPES 12.0g
精製水 1000ml
クエン酸3Na 100mg
TritonX−100(和光純薬工業株式会社) 0.2g
プロピジウムアイオダイド(シグマ) 0.2g
RO水 100ml
上記の血液にメイグリュンワルド染色を施した後、顕微鏡により目視を行った。白血球をリンパ球、単球、顆粒球に分類した。また、上記の血液を用いて、フローサイトメーターによるDNA量標準測定法にて、白血球のDNA量を測定した。得られた結果を、表1および図7に示す。
ポリオキシエチレン(16)オレイルエーテル 24.0g
N−ラウロイルサルコシン酸ナトリウム 1.5g
DL−メチオニン 20.0g
1N−NaOH 0.3g
NaCl 4.0g
式(VII)の色素 3.0mg
HEPES 12.0g
精製水 1000ml
クエン酸3Na 100mg
TritonX−100(和光純薬工業株式会社) 0.2g
プロピジウムアイオダイド(シグマ) 0.2g
RO水 100ml
DNA量標準測定法試薬1mlと上述の患者の骨髄液100μlを混合し、30分後にフローサイトメータで赤蛍光を測定した(光源:アルゴンイオンレーザー、波長:488nm)。得られた結果を、X軸に前方低角散乱光幅、Y軸に前方低角散乱光ピークをとった反応時間7秒の場合のスキャッタグラム(図11)、X軸に赤蛍光強度、Y軸に前方低角散乱光強度をとった反応時間7秒の場合のスキャッタグラム(図12)、X軸に側方散乱光強度、Y軸に赤蛍光強度をとった反応時間7秒の場合のスキャッタグラム(図13)、X軸に前方低角散乱光幅、Y軸に前方低角散乱光ピークをとった反応時間13秒の場合のスキャッタグラム(図14)、X軸に赤蛍光強度、Y軸に前方低角散乱光強度をとった反応時間13秒の場合のスキャッタグラム(図15)、X軸に側方散乱光強度、Y軸に赤蛍光強度をとった反応時間13秒の場合のスキャッタグラム(図16)に示す。
Claims (7)
- (1)血液学的試料を、赤血球を溶解させるための溶血剤によって処理する工程、
(2)処理された試料をフローサイトメータに導入して、少なくとも散乱光を測定する工程、
(3)前方散乱光幅の差を利用して、血小板凝集と、白血球とを分類する工程
からなる白血球の分類計数方法。 - 工程(3)において、前方散乱光幅の差と前方散乱光ピークの強度差を利用して、血小板凝集と、白血球とを分類する請求項1記載の白血球の分類計数方法
- さらに、(4)工程(3)で分類された白血球について側方散乱光の強度差を利用して、成熟白血球を少なくとも3つの亜集団に分類計数する工程、を備える請求項1または2記載の白血球の分類計数方法。
- 工程(1)において、血液試料を色素で処理する工程を含み、工程(2)において、さらに染色度合を測定する工程を含み、工程(4)において、工程(3)で分類された白血球について側方散乱光の強度差と染色度合の差を利用して、成熟白血球を少なくとも3つの亜集団に分類計数する請求項3記載の白血球の分類計数方法。
- 工程(1)において、血液試料を蛍光色素で処理する工程を含み、工程(2)において、さらに蛍光を測定する工程を含み、工程(4)において、工程(3)で分類された白血球について側方散乱光の強度差と蛍光の強度差を利用して、成熟白血球を少なくとも3つの亜集団に分類計数する請求項3記載の白血球の分類計数方法。
- 工程(1)が、血液学的試料を溶血剤で処理した後、溶血剤で処理した試料を蛍光色素で処理する工程を含む請求項5記載の白血球の分類計数方法。
- 血液学的試料を、赤血球を溶解させるための溶血剤によって処理する工程で調製された測定試料を通過させるオリフィス部と、オリフィス部に光を照射する光源と、オリフィス部から発せられた散乱光を受光する受光部とを備えたフローサイトメータと、
前方散乱光幅の差を利用して、血小板凝集と、白血球とを分類する工程により測定試料を解析する解析部と、を備えた白血球の分類計測装置。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2006319621A JP4354982B2 (ja) | 2002-06-24 | 2006-11-28 | 白血球の分類計数方法 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2002183259 | 2002-06-24 | ||
JP2006319621A JP4354982B2 (ja) | 2002-06-24 | 2006-11-28 | 白血球の分類計数方法 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2004515543A Division JP4324552B2 (ja) | 2002-06-24 | 2003-06-23 | 白血球の分類計数方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2007071890A true JP2007071890A (ja) | 2007-03-22 |
JP4354982B2 JP4354982B2 (ja) | 2009-10-28 |
Family
ID=37933419
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2006319621A Expired - Lifetime JP4354982B2 (ja) | 2002-06-24 | 2006-11-28 | 白血球の分類計数方法 |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP4354982B2 (ja) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2013522644A (ja) * | 2010-03-24 | 2013-06-13 | ベックマン コールター, インコーポレイテッド | 血液サンプルを分析するための方法およびシステム |
US10281458B2 (en) | 2015-09-14 | 2019-05-07 | Sysmex Corporation | Blood analyzer, blood analyzing method, and non-transitory computer-readable storage medium |
-
2006
- 2006-11-28 JP JP2006319621A patent/JP4354982B2/ja not_active Expired - Lifetime
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2013522644A (ja) * | 2010-03-24 | 2013-06-13 | ベックマン コールター, インコーポレイテッド | 血液サンプルを分析するための方法およびシステム |
US10281458B2 (en) | 2015-09-14 | 2019-05-07 | Sysmex Corporation | Blood analyzer, blood analyzing method, and non-transitory computer-readable storage medium |
Also Published As
Publication number | Publication date |
---|---|
JP4354982B2 (ja) | 2009-10-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP4324552B2 (ja) | 白血球の分類計数方法 | |
JP4976038B2 (ja) | 血液学的試料の測定方法 | |
EP1813942B1 (en) | Reagent for immature leukocyte analysis and reagent kit | |
JP4212827B2 (ja) | 骨髄液有核細胞自動分析方法 | |
JP3886271B2 (ja) | 赤芽球の分類計数用試薬及び分類計数方法 | |
JP4042925B2 (ja) | 幼若白血球の分類計数法 | |
EP1930723B1 (en) | Method for measuring biological sample and measuring apparatus therefor | |
US8455209B2 (en) | Reagent and reagent kit for analysis of immature leukocyte | |
US20140038179A1 (en) | Reagent and reagent kit for analysis of immature leukocyte | |
JP2002148261A (ja) | 異常細胞分類計数方法 | |
JP4806342B2 (ja) | 幼若白血球分析用試薬及び試薬キット | |
JP4354982B2 (ja) | 白血球の分類計数方法 | |
JP2006300962A (ja) | 白血球の分類計数方法 | |
JP3485436B2 (ja) | 網状赤血球測定用試薬及び測定方法 | |
JP5416232B2 (ja) | 血液学的試料の測定装置 | |
JP2012088336A (ja) | 骨髄芽球と血小板凝集との弁別方法及び弁別装置 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20090721 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20090730 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 4354982 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120807 Year of fee payment: 3 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120807 Year of fee payment: 3 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20150807 Year of fee payment: 6 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
EXPY | Cancellation because of completion of term |