10x Genomics Visium Spatial Transcriptome
Astaamaha
● Xallinta: 100 µM
● Dhexroorka Goobta: 55 µM
● Tirada dhibcaha: 4992
● Meesha la qaadayo: 6.5 x 6.5 mm
● Bar codeed kastaa waxaa lagu raray asal ka kooban 4 qaybood:
- dabada poly(dT) ee mRNA priming iyo cDNA synthesis
- Aqoonsiga Molecular Unique (UMI) si loo saxo eexashada
- Barcode meel bannaan
- Isku-xigta isku-xiran ee akhrinta qayb ahaan 1 tix-raaceed asal
● H&E midabaynta qaybaha
Faa'iidooyinka
●Adeeg hal joogsi ah: wuxuu isku daraa dhammaan khibradaha iyo tillaabooyinka ku saleysan xirfadda, oo ay ku jiraan qaybta cryo-sectioning, wasakhaynta, hagaajinta unugyada, barcodeing meel bannaan, diyaarinta maktabadda, isku xigxiga iyo bioinformatics.
● Koox farsamo oo xirfad sare leh: khibrad u leh in ka badan 250 nooc oo unug ah iyo 100+ nooc oo ay ku jiraan aadanaha, jiirka, naasleyda, kalluunka iyo dhirta.
●Cusboonaysiinta waqtiga-dhabta ah ee mashruuca oo dhan: iyadoo si buuxda loo xakameynayo horumarka tijaabada ah.
●Bioinformatics caadiga ah oo dhammaystiran:xirmada waxaa ku jira 29 falanqayn iyo 100+ tirooyin tayo sare leh.
●Falanqaynta xogta gaarka ah iyo muuqaalaynta: diyaar u ah codsiyada cilmi-baarista ee kala duwan.
●Falanqaynta wadajirka ah ee ikhtiyaarka ah oo leh hal unug oo mRNA isku xigxiga
Tilmaamaha
| Shuruudaha Tusaalaha | Maktabadda | Istaraatiijiyada isku xigxiga | Xogta lagu taliyay | Xakamaynta tayada |
| Saamballada cryo-ku-xidhan ee OCT, muunadaha FFPE (dhexroorka ugu fiican: qiyaastii. 6x6x6 mm3) 3 baloog halkii muunad | 10X Visium cDNA maktabadda | Illumina PE150 | 50K PE halkii buu akhriyaa (60Gb) | RIN>7 |
Faahfaahin dheeraad ah oo ku saabsan hagida diyaarinta muunada iyo socodka shaqada, fadlan xor u noqo inaad la hadasho aKhabiir BMKGENE
Socodka Shaqada Adeegga
Marxaladda diyaarinta muunada, tijaabada soo saarista RNA ee bilowga ah ayaa la sameeyaa si loo hubiyo in RNA tayo sare leh la heli karo.Marxaladda hagaajinta nudaha qaybaha waa la wasakheeyey oo la sawiray iyo xaaladaha sii daynta mRNA ee unugyada ayaa la hagaajiyey.Hab-maamuuska la hagaajiyay ayaa markaa lagu dabaqaa inta lagu jiro dhismaha maktabadda, oo ay ku xigto taxane iyo falanqaynta xogta.
Socodka shaqada ee dhamaystiran waxa uu ku lug leeyahay cusboonaysiinta wakhtiga dhabta ah iyo xaqiijinta macmiilka si loo joogteeyo wareegga jawaab celinta jawaab celinta, hubinta fulinta mashruuca.
Waxa ku jira falanqaynta soo socota:
Xakamaynta tayada xogta:
o Soo saarista xogta iyo qaybinta dhibcaha tayada
o Helitaanka hidda-wadaha halkii goobba
o daboolida unugyada
Falanqaynta muunada-gudaha:
o hodantinimada hidaha
o Goobo isku urursan, oo ay ku jiraan falanqaynta cabbirka oo la dhimay
o Falanqaynta tibaaxaha kala duwan ee u dhexeeya rucubyada: aqoonsiga hiddo-wadaha calaamadeeyayaasha
o Tilmaanta shaqaynaysa iyo kobcinta hiddo-wadaha calaamadaynta
Falanqaynta kooxaha dhexdooda
o Dib-u-isku-darka dhibcaha labada shayba (tusaale buka iyo xakamaynta) iyo dib-u-ururinta
o Aqoonsiga hidda-socodka calaamadeeyaha koox kasta
o Tilmaanta shaqaynaysa iyo kobcinta hiddo-wadaha calaamadaynta
o Muujinta kala duwanaanshaha isku koox ee u dhexeeya kooxaha
Falanqaynta muunada gudaha
Goobo ururin
Aqoonsiga hiddo-wadaha calaamadeeyaha iyo qaybinta meelaha bannaan
Falanqaynta kooxaha dhexdooda
Isku darka xogta labada kooxood iyo dib-u-kutlada
Hidde-sideyaasha kooxaha cusub
Sahami horumarka ay fududaysay BMKGene adeegiisa qoraalada qoraalka ee 10X Visium daabacaadyadan sifaysan:
Chen, D. iyo al.(2023) 'mthl1, suurtagalnimada Drosophila homologue ee naasleyda adhesion GPCRs, waxay ku lug leedahay falcelinta antitumor ee unugyada oncogenic ee duqsiyada', Tallaabooyinka Akadeemiyada Qaranka ee Sayniska ee Mareykanka ee Ameerika, 120(30), p.e2303462120.doi: /10.1073/pnas.2303462120
Chen, Y. iyo al.(2023) 'STEEL' waxa ay sahlaysa in si heersare ah loo kala saaro xogta qoraalka qoraalka ee spatiotemporal', Warbixin kooban oo ku saabsan Bioinformatics, 24(2), pp. 1-10.doi: 10.1093/BIB/BBAD068.
Liu, C. iyo al.(2022) 'A spatiotemporal atlas of organogenesis ee horumarinta ubaxa orchid', Cilmi-baarista Acids Nucleic, 50(17), bogga 9724-9737.doi: 10.1093/NAR/GKAC773.
Wang, J. iyo al.(2023) 'Isku-dhafka Farsamaynta Farsamaynta iyo Taxanaha Hal-Nucleus RNA waxay daaha ka qaadaysaa Xeeladaha Daawaynta ee suurtogalka ah ee Uterine Leiomyoma', Joornaalka Caalamiga ah ee Sayniska Nafleyda, 19(8), bogga 2515-2530.doi: 10.7150/IJBS.83510.
