I-10x ye-Genomics yeVisium ye-Spatial Transcriptome
Inkqubo yobuGcisa
Iimbonakalo
● Isisombululo: 100 µM
● I-Spot Diameter: 55 µM
● Inani lamabala: 4992
● Indawo yokubamba: 6.5 x 6.5 mm
● Indawo nganye enebhakhowudi ilayishwe ngeziqalo ezenziwe ngamacandelo ama-4:
-Ipoly(dT) umsila we-mRNA priming kunye ne-cDNA synthesis
-I-Unique Molecular Identifier (UMI) ukulungisa i-aplification bias
-Ibhakhowudi yendawo
- Ulandelelwano olubophelelayo lokufunda ngokuyinxenye 1 i-primer elandelelanayo
● Ukungcoliswa kwe-H&E yamacandelo
Iingenelo
●Inkonzo yokumisa enye: idibanisa onke amava kunye namanyathelo asekelwe kwizakhono, kuquka i-cryo-sectioning, i-staining, i-tissue optimization, i-barcoding yendawo, ukulungiswa kwethala leencwadi, ulandelelwano kunye ne-bioinformatics.
● IQela lobuGcisa elinezakhono eziphezulu: onamava ngaphezu kwe-250 yeentlobo zeethishu kunye ne-100+ yeentlobo ezibandakanya abantu, impuku, isilwanyana esanyisayo, intlanzi kunye nezityalo.
●Uhlaziyo lwexesha lokwenyani kwiProjekthi iyonke: ngolawulo olupheleleyo lwenkqubela phambili yovavanyo.
●I-Bioinformatics yoMgangatho oBanzi:iphakheji ibandakanya 29 uhlalutyo kunye 100+ amanani aphezulu.
●Uhlalutyo lwedatha oluLungisiweyo kunye nokuJonga: iyafumaneka kwizicelo ezahlukeneyo zophando.
●Uhlalutyo Oludityanisiweyo oluKhethekileyo kunye nolandelelwano lwe-mRNA yeseli enye
Iinkcukacha
| IiMfuno zeSampulu | Ithala leencwadi | Isicwangciso sokulandelelana | Idatha iyacetyiswa | Ulawulo lwemeko |
| Iisampulu ze-cryo ezifakwe kwi-OCT (Elona dayamitha ilungileyo: malunga ne-6x6x6 mm³) Iibhloko ezi-2 ngesampuli nganye | Ithala leencwadi le-10X Visium cDNA | Illumina PE150 | I-50K PE ifundeka kwindawo nganye ( 60Gb ) | RIN > 7 |
Ukufumana iinkcukacha ezithe vetshe malunga nesikhokelo sokulungiselela isampula kunye nokuhamba komsebenzi wenkonzo, nceda uzive ukhululekile ukuthetha nayeBMKGENE ingcaphephe
Ukuhanjiswa kwenkonzo
Kwinqanaba lolungiselelo lwesampulu, ulingo lokuqala lwe-RNA yesixa esikhulu lwenziwa ukuqinisekisa ukuba i-RNA yomgangatho ophezulu inokufumaneka. Kwinqanaba lokuphucula izicubu amacandelo abonakaliswe kwaye abonakaliswe kwaye iimeko ze-permeabilization zokukhululwa kwe-mRNA kwiithishu ziphuculwe. Iprothokholi ephuculweyo isetyenziswa ngexesha lolwakhiwo lwethala leencwadi, ilandelwa lulandelelwano kunye nohlalutyo lwedatha.
Inkonzo epheleleyo yokuhamba komsebenzi ibandakanya uhlaziyo lwexesha langempela kunye neziqinisekiso zabaxumi ukugcina i-loop yempendulo ephendulayo, ukuqinisekisa ukuphunyezwa kweprojekthi kakuhle.
Ibandakanya uhlalutyo olulandelayo:
Ulawulo Lomgangatho Wedatha:
o Isiphumo sedatha kunye nokuhanjiswa kwamanqaku asemgangathweni
o Ukufunyanwa kwemfuza kwindawo nganye
o Ukugquma kwezicubu
Uhlalutyo lwesampulu yangaphakathi:
o Ubutyebi bemfuza
o Ukuhlanganiswa kwamabala, kubandakanywa uhlalutyo oluncitshisiweyo lobungakanani
o Uhlalutyo lwentetho eyohlukileyo phakathi kwamaqela: ukuchongwa kofuzo lokumakisha
o Inkcazo esebenzayo kunye nokutyetyiswa kweejene zokumakisha
Uhlalutyo lwamaqela
o Ukudityaniswa kwakhona kwamabala kuzo zombini iisampulu (umzekelo, izifo kunye nolawulo) kunye nokuphinda kuhlangane
o Ukuchongwa kofuzo olumakishayo kwiqela ngalinye
o Inkcazo esebenzayo kunye nokutyetyiswa kweejene zokumakisha
o Umahluko Ubonakaliso lweqela elifanayo phakathi kwamaqela
Uhlalutyo lwesampulu yangaphakathi
Ukuhlanganiswa kwamabala
Ukuchongwa kofuzo lophawu kunye nokuhanjiswa kwendawo
Uhlalutyo lwamaqela
Indibaniselwano yedatha esuka kumaqela omabini kunye neqela kwakhona
Iijeni zokumakisha zamaqela amatsha
Phonononga inkqubela phambili eququzelelwa yinkonzo ye-BMKGene ye-spatial transcriptomics ngo-10X Visium Kolu papasho lubonakalisiweyo:
Chen, D. et al. (2023) 'mthl1, i-Drosophila homologue enokubakho ye-mammalian adhesion GPCRs, ibandakanyeka kwiimpendulo ze-antitumor kwiiseli ze-oncogenic ezifakwe kwiimpukane',Iinkqubo zeZiko leSizwe leSayensi lase-United States of America, 120(30), iphe. e2303462120. doi: /10.1073/pnas.2303462120
Chen, Y. et al. (2023) 'I-STEEL yenza i-high-resolution delineation ye-spatiotemporal transcriptomic data',Amagqabantshintshi kwiBioinformatics, 24(2), iphepha 1–10. doi: 10.1093/BIB/BBAD068.
U-Liu, uC. et al. (2022) 'I-atlas ye-spatiotemporal ye-organogenesis ekuphuhliseni iintyatyambo ze-orchid',Uphando lweNucleic Acids, 50(17), iphepha 9724-9737. doi: 10.1093/NAR/GKAC773.
Wang, J. et al. (2023) 'Ukudibanisa i-Spatial Transcriptomics kunye ne-Single-nucleus RNA Sequencing ityhila iiNdlela eziSebenzayo zonyango lwe-Uterine Leiomyoma',Ijenali yeHlabathi yeSayensi yebhayoloji, 19(8), iphepha 2515–2530. doi: 10.7150/IJBS.83510.
