DK2304020T3 - Method and apparatus for sorting cells - Google Patents
Method and apparatus for sorting cells Download PDFInfo
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- DK2304020T3 DK2304020T3 DK09772915.6T DK09772915T DK2304020T3 DK 2304020 T3 DK2304020 T3 DK 2304020T3 DK 09772915 T DK09772915 T DK 09772915T DK 2304020 T3 DK2304020 T3 DK 2304020T3
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Classifications
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- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
-
- 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—Electro-optical investigation, e.g. flow cytometers
- G01N15/1404—Fluid conditioning in flow cytometers, e.g. flow cells; Supply; Control of flow
-
- G01N15/1409—
-
- G01N15/149—
Claims (13)
1
1. Fremgangsmåde til selektion af et første sæt celler fra en cellepopulation omfattende et første sæt celler og et andet sæt celler, hvilken fremgangsmåde omfatter: mærkning af cellepopulationen, således at det første sæt celler kan adskilles fra det andet sæt celler; tilvejebringelse af en første strømningssti (110) med en distal ende, en proksimal ende, et undersøgelsesområde (128) placeret mellem den proksimale ende og den distale ende, og en strøm ningsakse; dannelse af en indkapslingsstrøm ning (1 1 6) af et indkapslingsfluid gennem den første strømningssti, idet indkapslingsstrømningen bevæger sig mod den proksimale ende med en første strømningshastighed; injektion ind i indkapslingsstrømningen, ved et punkt opstrøms i forhold til den proksimale ende, af en prøvestrømning (120) omfattende cellepopulationen, idet prøvestrømningen indledningsvis har en anden strømningshastighed, som er mindre end den første strøm ningshast ighed; tilvejebringelse af en excitationsenergikilde, idet energien, som udsendes fra excitationsenergikilden, virker på individuelle celler, når de passerer gennem undersøgelsesområdet og forårsager emission eller transmission af sekundær stråling fra cellerne; anvendelse af en objektivlinse (142) med en optisk akse (144), som generelt er koaksialt placeret langs med strømningsaksen (130) til at fokusere den sekundære stråling fra de individuelle celler, når cellerne passerer gennem undersøgelsesområdet; detektion af den fokuserede sekundære stråling fra de individuelle celler; bestemmelse af, fra den detekterede sekundære stråling, hvorvidt de individuelle celler er i det første sæt eller det andet sæt; og selektion af celler, som er bestemt til at være i det første sæt.
2. Fremgangsmåde ifølge krav 1, hvor energien, som udsendes fra excitationsenergikilden, passerer gennem objektivlinsen.
3. Fremgangsmåde ifølge krav 1 eller krav 2, hvori selektion af celler, som er bestemt til at være i det første sæt, omfatter anvendelse af en differentieringsenergikilde til bestråling af celler, som ikke er bestemt til at være i det første sæt.
4. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 3, hvori selektion af celler, som er bestemt til at være i det første sæt, omfatter derivatisering, aflivning, beskadigelse, modificering, sprængning eller fragmentering af celler, som ikke er bestemt til at være i det første sæt.
5. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 4, som yderligere omfatter tilvejebringelse af en anden strømningssti på tværs af strømningsaksen og placeret ved den proksimale ende af den første strømningssti, således at cellerne, efter passage gennem undersøgelsesområdet og når de når den proksimale ende af den første strømningssti, bevæger sig ind i den anden strømningssti og bort fra undersøgelsesområdet.
6. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 5, hvori tilvejebringelse af en strømningssti yderligere omfatter tilvejebringelse af en strømningssti, hvoraf i det mindste en del er dannet af et materiale med et brydningsindeks i intervallet 1,30 til 1,40 inklusive.
7. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 6, som yderligere omfatter justering af brydningsindekset af en opløsning indeholdende cellepopulationen, således at opløsningens brydningsindeks er inden for 0,02 af brydningsindekset for det materiale, som danner undersøgelsesregionen af strømningsstien.
8. Apparat til detektion og selektiv ændring af en ønsket sub-population af celler i en population af prøveceller, hvilket apparat omfatter: en fluidstrømningssti (110) med: en første strømningssektion, der har en strømningsakse (130), og en anden strømningssektion, idet den første og anden strømningssektion skærer hinanden ved en måleende af den første strømningssektion; et undersøgelsesområde (128) placeret ved eller nær måleenden af den første strømningssektion; et indkapslingsfluidindløb (114) i fluidstrømningsforbindelse med fluidstrømningsstien; et prøveindløb (122) i fluidstrømningsforbindelse med fluidstrømningsstien; en objektivlinse (142) med et nominelt fokuspunkt og en optisk akse (144) og placeret ved måleenden af den første strømningssektion, idet objektivlinsen er placeret langs med den første strømningssektion, således at det nominelle fokuspunkt er langs strømningsaksen og i undersøgelsesområdet, og således, at den optiske akse generelt er koaksialt placeret langs med strømningsaksen; en detektor placeret til at detektere lys fokuseret af objektivlinsen; en logikrutine kommunikativt koblet til detektoren, og som kan betjenes til bestemmelse af, hvorvidt en celle i populationen af prøveceller er en af den ønskede sub-population af celler, og som endvidere kan betjenes til udsendelse af et signal baseret på bestemmelsen af, hvorvidt cellen er en af den ønskede sub-population af celler; og en styrbar energikilde, som er kommunikativt koblet til logikrutinen og kan betjenes til selektivt at ændre enten celler i den ønskede sub-population af celler eller celler, som ikke er i den ønskede sub-population af celler ifølge i det mindste signaludgangen fra logikrutinen.
9. Apparat ifølge krav 8, som yderligere omfatter en excitationsenergikilde.
10. Apparat ifølge krav 9, hvori excitationsenergikilden omfatter en dæmper med den styrbare energikilde som et input.
11. Apparat ifølge enten krav 9 eller krav 1 0, hvori energien udsendt fra den styrbare energikilde passerer gennem objektivlinsen.
2. Apparat ifølge et hvilket som helst af krav 8 til 11, som yderligere omfatter: et legeme, hvori strømningsstien er dannet; og en rille, som danner den anden strømningssektion i en første overflade af legemet, hvori måleenden af den første strømningssektion skærer rillen ved den første overflade.
3. Apparat ifølge et hvilket som helst af krav 8 til 11, som yderligere omfatter: et legeme, hvori strømningsstien er dannet; en første indre kanal gennem legemet, som strækker sig fra en første overflade af legemet til et punkt i legemet og danner den første strømningssektion; og en anden indre kanal gennem legemet, som strækker sig fra punktet i legemet til en anden overflade af legemet.
14. Apparat ifølge enten krav 12 eller krav 13, hvori legemet er dannet af et materiale med et brydningsindeks mellem 1,30 og 1,40 inklusive.
15. Apparat ifølge krav 14, hvori materialet omfatter en amorf perfluorpolymer, en amorf fluorpolymer eller en pertluoralkoxypolymer.
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EP2304020A4 (en) | 2017-10-11 |
MX2011000182A (es) | 2011-08-03 |
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WO2010001254A3 (en) | 2010-04-01 |
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WO2010001254A2 (en) | 2010-01-07 |
US20120038914A1 (en) | 2012-02-16 |
UA102854C2 (ru) | 2013-08-27 |
CN102119212B (zh) | 2014-08-20 |
EP2304020B1 (en) | 2018-11-07 |
EP2304020A2 (en) | 2011-04-06 |
UA115521C2 (uk) | 2017-11-27 |
RU2011102290A (ru) | 2012-08-10 |
US8467040B2 (en) | 2013-06-18 |
CA2728289A1 (en) | 2010-01-07 |
CN102119212A (zh) | 2011-07-06 |
BRPI0914919A2 (pt) | 2015-08-25 |
RU2520848C2 (ru) | 2014-06-27 |
US8004661B2 (en) | 2011-08-23 |
US20090325217A1 (en) | 2009-12-31 |
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