Imikhiqizo

I-Single-nucleus RNA Sequencing

I-Single-nucleus RNA Ukulandelanisa Isithombe Esifakiwe

Ukuthuthuka ekuthwebuleni iseli elilodwa kanye nezindlela zokwakha umtapo wolwazi ngamunye okuhlanganiswe nokulandelana komphumela ophezulu kuvumela izifundo zesisho sofuzo ngesisekelo seseli-by-cell.Inika amandla ukuhlaziya okujulile nokuphelele kwesistimu kumaseli ayinkimbinkimbi, lapho igwema khona kakhulu ukufihla ukuhlukahluka kwawo ngokuthatha isilinganiso sawo wonke amaseli.

Kodwa-ke, amanye amangqamuzana awafanele ukwenziwa abe yi-single-cell ukumiswa, ngakho-ke ezinye izindlela zokulungiselela isampula - ukukhishwa kwe-nucleus ezicutshini ziyadingeka, okungukuthi, i-nucleus ikhishwa ngokuqondile kumathishu noma ingqamuzana futhi ilungiselwe ukumiswa kwe-nucleus eyodwa ukuze kusetshenziswe eyodwa- ukulandelana kweseli.

I-BMK ihlinzeka ngesevisi yokulandelana kwe-RNA eyi-10× Genomics ChromiumTM.Le nsizakalo isetshenziswe kakhulu ezifundweni eziphathelene nezifo, njengokuhlukaniswa kwamaseli omzimba, i-tumor heterogeneity, ukuthuthukiswa kwezicubu, njll.

I-Spatial transcriptome chip: 10× Genomics

Ipulatifomu: I-Illumina NovaSeq Platform

Xhumana nathi

Imininingwane Yesevisi

I-Bioinformatics

Imiphumela yedemo

Isimiso sobuchwepheshe

Ukuhlukaniswa kwe-nuclei kufinyelelwa yi-10 × i-Genomics Chromium TM, equkethe uhlelo lwe-microfluidics lwesiteshi esiyisishiyagalombili ngokuwela kabili.Kulolu hlelo, ubuhlalu bejeli obunamabhakhodi kanye ne-primer, ama-enzyme kanye ne-nucleus eyodwa kufakwe e-nanoliter-sized oil drop drop, ekhiqiza i-Gel Bead-in-Emulsion (GEM).Uma i-GEM seyakhiwe, i-cell lysis kanye nokukhululwa kwamabhakhodi kwenziwa ku-GEM ngayinye.I-mRNA ibhalwe ngokuhlehlayo ibe ama-molecule e-cDNA anamabhakhodi angu-10× kanye ne-UMI, angaphansi kokwakhiwa komtapo wolwazi wokulandelanisa okujwayelekile.

Izicubu azilungele ukulungiswa kweseli elilodwa

Iseli / Izicubu	Isizathu
Susa izicubu eziqandisiwe	Ayikwazi ukuthola izinhlangano ezintsha noma ezilondolozwe isikhathi eside
Amaseli emisipha, iMegakaryocyte, Amafutha…	Ububanzi beseli bukhulu kakhulu ukuthi ungangena entweni
Isibindi...	Intekenteke kakhulu ukuthi ingaphuka, ayikwazi ukuhlukanisa iseli elilodwa
Iseli ye-Neuron, Ubuchopho...	Ukuzwela okwengeziwe, kulula ukucindezela, kuzoshintsha imiphumela yokulandelana
Ama-pancreas, i-thyroid ...	Icebile kuma-endogenous enzymes, athinta ukukhiqizwa kokumiswa kweseli elilodwa

I-nucleus eyodwa vs iseli elilodwa

I-nucleus eyodwa	Iseli elilodwa
Ububanzi beseli obungenamkhawulo	Ububanzi beseli: 10-40 μm
Impahla ingaba izicubu eziqandisiwe	Impahla kufanele ibe izicubu ezintsha
Ingcindezi ephansi yamaseli afriziwe	Ukwelashwa ngama-enzyme kungase kubangele ukusabela kokucindezeleka kweseli
Awekho amangqamuzana abomvu egazi okudingeka akhishwe	Amangqamuzana egazi abomvu kudingeka akhishwe
Nuclear iveza bioinformation	Iseli lonke liveza ulwazi lwezinto eziphilayo

Imininingwane Yesevisi

Umtapowolwazi

Isu lokulandelanisa

Ivolumu Yedatha

Izidingo Zesampula

Izicubu

10× Ilabhulali ye-Genomics eyodwa-nuclei

10x Genomics -Illumina PE150

100,000 ukufundwa/cell approx.100-200 Gb

Inombolo yeseli: >2×10⁵

Iseli conc.ku-700-1,200 iseli/μL

≥ 200 mg

Ukuze uthole imininingwane eyengeziwe ngomhlahlandlela wokulungiselela isampula kanye nokuhamba komsebenzi wesevisi, sicela ukhululeke ukukhuluma no-aUchwepheshe we-BMKGENE