TR202018906A2 - Tek hücre jel elektroforezi̇ akiş si̇tometri̇ - Google Patents

Tek hücre jel elektroforezi̇ akiş si̇tometri̇

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Publication number
TR202018906A2
TR202018906A2 TR2020/18906A TR202018906A TR202018906A2 TR 202018906 A2 TR202018906 A2 TR 202018906A2 TR 2020/18906 A TR2020/18906 A TR 2020/18906A TR 202018906 A TR202018906 A TR 202018906A TR 202018906 A2 TR202018906 A2 TR 202018906A2
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cells
electro
single cell
slide
flow cytometry
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TR2020/18906A
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English (en)
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Güner Utku
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Trakya Ueniversitesi Rektoerluegue
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Priority to TR2020/18906A priority Critical patent/TR202018906A2/tr
Priority to PCT/TR2021/051177 priority patent/WO2022115076A1/en
Priority to US18/253,932 priority patent/US20230415143A1/en
Priority to EP21876723.4A priority patent/EP4028817A4/en
Publication of TR202018906A2 publication Critical patent/TR202018906A2/tr

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    • G—PHYSICS
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    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/0241—Drop counters; Drop formers
    • B01L3/0268—Drop counters; Drop formers using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44756—Apparatus specially adapted therefor
    • G01N27/44786—Apparatus specially adapted therefor of the magneto-electrophoresis type
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009—Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G01N35/1016—Control of the volume dispensed or introduced
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1081—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0636—Focussing flows, e.g. to laminate flows
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/069—Absorbents; Gels to retain a fluid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/043—Moving fluids with specific forces or mechanical means specific forces magnetic forces
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/282—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on with mapping; Identification of areas; Spatial correlated pattern
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1034—Transferring microquantities of liquid
    • G01N2035/1041—Ink-jet like dispensers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44704—Details; Accessories

Abstract

Buluş, SCGE analizlerinde hücrelerin tek tek, belirli hız ve doğrultuda lam üzerindeki agaroz katmanındaki üzerine dağılması ile ilgilidir.

Description

TARIFNAME TEK HÜCRE JEL ELEKTROFOREZI AKlS SITOMETRI TEKNIK ALAN Bulus, SCGE analizlerinde hücrelerin tek tek, belirli hTz ve dogrultuda lam üzerindeki agaroz katman ndaki üzerine dag Imasl ile ilgilidir. TEKNIGIN BILINEN DURUMU Teknigin bilinen durumunda "Single Cell Gel Electrophoresis" (SCGE) teknigi DNA kiüllarii tespiti için kullanüan hassas, hîljve ucuz bir yöntemdir. SCGE teknigi "Comet Assay" ya da "Microgel Electrophoretic Technique" olarak da adlandmümaktadîl Söz konusu teknikte kullan Jhn birçok yöntem olmas îla kars El, tüm yöntemlerde, kullanüan hücreler pipetle düsük kaynama dereceli agaroz (LMP) ile (37 oC) karLsltLudarak lam üzerindeki agaroz katmanl üzerine aktar lln Bu islemi günümüze kadar otomasyon gerçeklestiren bir cihaz yoktur. Hücrelerin tek tek ve düzenli olarak LMP üzerine aktarllamaz hücreler rasgele lam üzerindeki agaroz katmanrdaglinaktadî, hücrelerin lam üzerindeki agaroz katmanEldaki dag [Iînükontrol edilememektedir. Hücrelerin ne kadar sklkla ne kadar derinligi hangi desen yerlestirilecegi bilinemez. Bu sorunlar SCGE analizlerinde sßmlama getirmektedir. SCGE analizlerinde LMA üzerine hücreler basitçe pipetle dag EUR. Hücrelerin lam üzerindeki agaroz katmanEidaki üzerine ne kadar aralkla ve nas[[ dagmEn gösterecegi öngörülemez. Hücrelerin çalkalanmas Lbu sorunu kesin ve emin bir biçimde çözmez. Dengeli uygun sLklUstla hücre dag llml olmamas ISCGE analizinde önemli bir sorunudur. Bu sorun nedeniyle hücre dagllml luygun olmayan preparatlar ve/ veya preparatlar n bir kldml lSCGE analizlerinde degerlendirilineye allîlamaz. BULUSUN KISA AÇIKLAMASI Söz konusu olan bulus, yukarIla yer alan dezavantajlarEortadan kaldîmak ilgili teknik alana yeni avantajlar getirmek üzere LMA (agaroz) üzerine hücrelerin uygun derinlikte ve desende dag lmas It lsaglayan bir cihaz ile ilgilidir. Bu bulus hücreleri hidrodinamik odaklama tek tek LMA üzerine saglar, elektro-manyetik yönlendirme gücüne baglFolarak LMA"nlîi ne kadar derin girecegi belirlenir. Ayröa Elektro- manyetik yönlendirme hücrenin LMA üzerinde istenen düzene bagll l olarak dagllml gerçeklestirilir. Kullanii istemine baglj olarak farklü desen ve derinlikte damlacEk ve hücre dagtlEnD gerçeklestirilmektedir. Söz konusu bulus, SCGE (tek hücreli jel elektroforezi) analizlerinde lam üzerindeki agaroz katmanîtdaki üzerinde istenen SÜMER, desende ve uygun derinlikte hücre yerlestirmektedir. Aerda bulus çok az sayLda hücre ile SCGE analiz yap Llmas Lna imkan sunmaktadii Sistem SCGE analizlerin hlzll, tekrarlanabilir olmas nl ldaha az is gücü ile güvenilir yap llnas îia olanak tan ü SEKILLERIN KISA AÇIKLAMASI Söz konusu bulusun daha iyi anlasürnasEiçin sekiller ve sekillerle ilgili açklamalar asagEla yer almaktadi. Sekil 1 Elektro-manyetik odaklama sistemi Sekil 2 Elektro-manyetik yönlendirme ile hareket eden damlacEk nozulu Sekil 3 Elektro-manyetik yüklendirme için sarng Sekil 4 Lam hareketi saglayan elektrik motor sistem Sekil 5 Cihaz n temel tasar In | REFERANS NUMARALARI Söz konusu bulusun daha iyi anlaSJInas :için unsurlar ve unsurlara ait referans numaralarE asag Zia yer almaktad î. 1 Elektro-manyetik odaklama sistemi 2 Damlac k nozulu 3 Elektrik sarg 511 l 4 Elektrik motor Temel cihaz 6 Manyetik malzeme 7 Nozulun olas Fhareket yönleri 8 Esnek baglantj 9 Nozul ucu Damlac 11 l 1 Bak] sarg 3 12 LMA Lam 13 Ayak 14 Sonsuz vida Kontrol sistemi BULUSUN DETAYLI AÇIKLAMASI Bu detayljaç klamada bulus konusu yenilik sadece konunun daha iyi anlas Elmasiia yönelik hiçbir si IllayB :Etki olusturmayacak örneklerle aç [klanmaktad E. Söz konusu bulus, tek hücre jel elektroforezi akß sitometri ile ilgilidir. Bulus, (1) Elektro- manyetik odaklama sistemi, (1) elektro-manyetik yönlendirme ile hareket eden damlacEk nozulu (2), elektro-manyetik yüklendirme için elektro magnetik alan olusturan elektrik sargE (3), lam hareketi saglayan elektrik motoru (4) elektro-manyetik odaklama sistemi ve elektrik motoru kontrol eden kontrol sistemini (15) içermektedir. Söz konusu bulus, uygun sIlaklEta LMA içinde bulunan hücreler elektro-manyetik odaklama ile elektrik alan içinde yönlendirerek elektrik motorla düsük hîda sürekli hareket eden lam üzerine püskürtmektedir. Hücrelerin dagLlLmLh/e lam içindeki derinligi elektrik motor hLanaj hareketli nozulun elektro-manyetik odaklama bagl lhareketine nozuldaki püskürtme hlzlna, elektro-manyetik odaklama saglayan elektrik sarglar Il Il gönderilen ak m miktarl .ve aklm yüklerine bag] `olarak degisir ve bu yolla istenen desen, sklfk ve derinlikte düzenli ve tekrar edilebilir hücre dagll m Saglan It Yukarîla yer alan detayljaçiklamalara istinaden söz konusu olan bulus, LMA üzerine hücrelerin uygun derinlikte ve desende dag EllnasEiEsaglayan sistem olup özelligi; o Nozuldan çikan hücrelerin dag Im] desenini belirleyen, elektro-manyetik yönlendirme ile nozul hareketine baglEolarak hücreleri farklEyönlerde, x ve y koordinatlarülda, istenen derinlikte ve istenen hâda nozuldan tek tek damlac Elar göndermesini saglayan esnek baglantEKS) içeren, LMA içinde bulunan hücreleri elektrik alan içinde yönlendirerek elektrik motorla düsük hâda sürekli hareket eden lam üzerine püskürtülmesini saglayan elektro-manyetik odaklama sistemi (1), o Hareket etmesinde kullanlhiasj için üzerinde manyetik malzeme (6) olan, damlac Jîlarîi çüîßmîsaglayan elektro-manyetik yönlendirme ile hareket eden x ve y eksenlerinde hareket etmesini saglayan esnek baglant]D(8) en az bir adet damlacüg o Damlacl& nozulunun (2) elektro-manyetik yönlendirme hareketini saglayan, elektro- manyetik odaklama için farklüakîn yönü ve akEh miktarEiEbelirleyerek elektro- manyetik yönlendirme saglayan elektrik sarg@:(3), o LMA tasgian lamîi hareket etmesini saglayan, sonsuz Vida ve elektrik motoiu ile lamln hareket yönünü ve hlzlnl belirleyerek hücre dag llm deseni düzenlenmesini saglayan en az bir adet elektrik motor (4), - Kulland. listegine uygun olarak elektro-manyetik odaklama (l) hlZlnln ayarlanmasl,l elektro-manyetik odaklama sargllarlnln voltaj ve aklm yönlerinin ve LMA taslyan lamühareket ettiren elektrik motoru (4) denetlenmesini (hareket hâüve hareket yönü olarak) saglayan kontrol sistemi (5) unsurlarmüçermesidir. Bulusun diger düzenlemeleri; LMA üzerine hücrelerin uygun (istenen) derinlikte ve desende dagEmasEiEsaglayan cihaz olup özelligi yukarEla bahsi geçen sistemi içermesidir. Hücre dagllllînüi düzenlemesi x ve y koordinatlarüida saglayan esnek baglantryr içermesidir. Bahsi geçen elektrik motorunu (4)lam uzunlugu boyunca, her iki yönde, sonsuz Vida ile çok küçük ad mlarla hassas kontrol edilmesini saglayan kontrol sistemini (5) içermesidir. Bahsi geçen elektrik motorunun (4) step motor olmasEli. Bahsi geçenHücrelerin dagJEij ve lam içindeki derinligi elektrik motor hüßia, hareketli nozulun elektro-manyetik odaklama bagljhareketine nozuldaki püskürtme hîia, elektro-manyetik odaklama saglayan elektrik sargührßm gönderilen akin miktar Eve akil yüklerine baglîblmasîli TR TR TR
TR2020/18906A 2020-11-24 2020-11-24 Tek hücre jel elektroforezi̇ akiş si̇tometri̇ TR202018906A2 (tr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
TR2020/18906A TR202018906A2 (tr) 2020-11-24 2020-11-24 Tek hücre jel elektroforezi̇ akiş si̇tometri̇
PCT/TR2021/051177 WO2022115076A1 (en) 2020-11-24 2021-11-10 Single cell gel electrophoresis flow cytometry
US18/253,932 US20230415143A1 (en) 2020-11-24 2021-11-10 Single cell gel electrophoresis flow cytometry
EP21876723.4A EP4028817A4 (en) 2020-11-24 2021-11-10 SINGLE CELL GEL ELECTROPHORESIS FLOW CYTOMETRY

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TR2020/18906A TR202018906A2 (tr) 2020-11-24 2020-11-24 Tek hücre jel elektroforezi̇ akiş si̇tometri̇

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US (1) US20230415143A1 (tr)
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WO (1) WO2022115076A1 (tr)

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JP4572022B2 (ja) * 2000-07-06 2010-10-27 光夫 板倉 平板ゲル電気泳動装置用試料導入装置およびその試料導入方法
CN201662556U (zh) * 2010-03-09 2010-12-01 浙江大学 一种用于单细胞凝胶电泳试验的成套装置
AU2012318963B2 (en) * 2011-09-25 2016-02-11 Labrador Diagnostics Llc Systems and methods for multi-analysis
WO2015079048A1 (en) * 2013-11-29 2015-06-04 Ge Healthcare Bio-Sciences Ab Electrophoresis system
EP3265796B1 (de) * 2015-03-06 2022-11-09 4D Lifetec AG Gelelektrophorese-system für einzelzell-gelelektrophorese
WO2019210243A1 (en) * 2018-04-26 2019-10-31 The Regents Of The University Of California Methods and devices using individually addressable microgel electrophoresis lane synthesis for bioassays

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