I-BMKMANU S1000 i-Spatial Transcriptome
BMKMANU S1000 Spatial Transcriptome Technical Scheme
Iimbonakalo
● Isisombululo: 5 µM
● I-Spot Diameter: 2.5 µM
● Inani lamabala: malunga nezigidi ezi-2
● 3 iifomathi ezinokubakho zendawo yokubamba: 6.8 mm * 6.8 mm, 11 mm * 11 mm okanye 15 mm * 20 mm
● Iso ngalinye elinebhakhowudi lilayishwa ngezinto zokuqala ezenziwe ngamacandelo ama-4:
i-poly(dT) umsila we-mRNA priming kunye ne-cDNA synthesis
Isichongi se-Molecular Eyodwa (UMI) ukulungisa ukuthambekela kokukhulisa
Ibhakhowudi yendawo
Ulandelelwano olubophelelayo lokufunda ngokuyinxenye 1 i-primer yolandelelwano
● I-H&E kunye ne-fluorescent staining yamacandelo
● Ukukwazi ukusebenzisaitekhnoloji yokwahlulahlula iiseli: ukuhlanganiswa kwe-H & E staining, i-fluorescent staining, kunye nokulandelelana kwe-RNA ukumisela imida yeseli nganye kunye nokwabela ngokuchanekileyo ukubonakaliswa kofuzo kwiseli nganye.
Izinto ezilungileyo ze-BMKMANU S1000
●ISigqibo esisezantsi kweselula: Indawo nganye yokubanjwa iqulathe >2 yezigidi ezisi-2 zeNdawo ezineBarcoded ezisisithuba esinobubanzi obuyi-2.5 µm kunye nesithuba se-5 µm phakathi kwamaziko amabala, oko kuvumela uhlalutyo lwe-transcriptome yendawo kunye nesisombululo se-sub-cellular (5 µm).
●Uhlalutyo lweSisombululo esikwinqanaba eliphezulu:Uhlalutyo oluguquguqukayo lwamanqanaba amaninzi ukusuka kwi-100 μm ukuya kwi-5 μm ukusombulula iimpawu zezicubu ezahlukeneyo kwisisombululo esisiso.
● Ukubanakho ukusebenzisa “Ezintathu kwisilayidi esinye” ITekhnoloji yokwahlulwa kweeseli:Ukudibanisa i-fluorescence staining, i-H&E staining, kunye nolandelelwano lwe-RNA kwisilayidi esinye, i-algorithm yethu yokuhlalutya "ezintathu-kwinye" ixhobisa ukuchongwa kwemida yeeseli kwi-transcriptomics elandelayo esekwe kwiseli.
●Iyahambelana neePlatifomu zolandelelwano ezininzi: Zombini i-NGS kunye nolandelelwano olude lokufunda luyafumaneka.
●Uyilo oluguquguqukayo lwe-1-8 Indawo yokuThatha esebenzayo: Ubungakanani bendawo yokubamba buguquguqukayo, kunokwenzeka ukusebenzisa iifomathi ze-3 (6.8 mm * 6.8 mm., 11 mm * 11 mm kunye ne-15 mm * 20 mm)
●Inkonzo yokumisa enye: Idibanisa onke amava kunye namanyathelo asekelwe kwizakhono, kubandakanywa i-cryo-sectioning, i-staining, i-tissue optimization, i-barcoding yendawo, ukulungiswa kwethala leencwadi, ukulandelelana, kunye ne-bioinformatics.
●I-Bioinformatics eBanzi kunye nokubonwa kweZiphumo ezisebenziseka lula:iphakheji ibandakanya 29 uhlalutyo kunye 100+ amanani aphezulu, idityaniswe nokusetyenziswa inhouse eziphuhliswe isoftwe ukuze ubone kwaye wenze ngokusesikweni ukwahlula iseli kunye nebala ukuhlanganisa.
●Uhlalutyo lwedatha oluLungisiweyo kunye nokuJonga: iyafumaneka kwizicelo ezahlukeneyo zophando
●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 yonke: ngolawulo olupheleleyo lwenkqubela phambili yovavanyo.
●Uhlalutyo Oludityanisiweyo oluKhethekileyo kunye nolandelelwano lwe-mRNA yeseli enye
IiNgcaciso zeNkonzo
|
Isampulu Iimfuno
| Ithala leencwadi |
Isicwangciso sokulandelelana
| Idatha iyacetyiswa | Ulawulo lwemeko |
| Iisampuli ze-cryo ezifakwe kwi-OCT, iibhloko ezi-3 ngesampuli | S1000 cDNA ilayibrari | Illumina PE150 (amanye amaqonga akhoyo) | I-100K PE ifundwa nge-100 uM ( 60-150 Gb ) | I-RIN>7 |
Ukufumana iinkcukacha ezithe vetshe malunga nesikhokelo sokulungiselela isampula kunye nokuhamba komsebenzi wenkonzo, nceda uzive ukhululekile ukuthetha nayeBMKGENE ingcaphephe
Ukuhamba komsebenzi weNkonzo
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.
Idatha eveliswe yi-BMKMANU S1000 ihlalutywa kusetyenziswa i-software "BSTMatrix", eyenziwe ngokuzimeleyo yi-BMKGENE, ivelisa i-Gene Expression Matrix. Ukusuka apho, kuveliswa ingxelo esemgangathweni equka ulawulo lomgangatho wedatha, uhlalutyo lwangaphakathi lwesampuli kunye nohlalutyo lwamaqela.
● Ulawulo Lomgangatho Wedatha:
Isiphumo sedatha kunye nokuhanjiswa kwamanqaku asemgangathweni
Ukufunyanwa kwemfuza kwindawo nganye
Ukugquma kwezicubu
● Uhlalutyo lwesampulu yangaphakathi:
Ubutyebi bemfuza
Ukuhlanganiswa kwamabala, kubandakanywa uhlalutyo oluncitshisiweyo lobungakanani
Uhlalutyo lwentetho eyohlukileyo phakathi kwamaqela: ukuchongwa kofuzo lokumakisha
Isichasiselo esisebenzayo kunye nokutyetyiswa kweejene zokumakisha
● Uhlalutyo lwamaqela ngamaqela:
Ukudityaniswa kwakhona kwamabala kuzo zombini iisampulu (umzekelo, izifo kunye nolawulo) kunye nokuphinda kuhlangane
Ukuchongwa kofuzo lwamakisha kwiqela ngalinye
Isichasiselo esisebenzayo kunye nokutyetyiswa kweejene zokumakisha
Umahluko Ubonakaliso lweqela elifanayo phakathi kwamaqela
Ukongeza, i-BMKGENE iphuhliswe "i-BSTViewer" sisixhobo esisebenziseka lula esenza ukuba umsebenzisi abe nomfanekiso ngqondweni wembonakalo yemfuza kunye nokudibanisa amabala kwizisombululo ezahlukeneyo.
I-BMKGene iphuhlise isoftware yokujonga umsebenzisi
BSTViewer indawo yokudibanisa kwisisombululo esinamanqanaba amaninzi
BSTCellViewer: ngokuzenzekelayo kunye nokwahlulwa kweseli ngesandla
Uhlalutyo lwesampulu yangaphakathi
Ukuhlanganiswa kwendawo:
Ukuchongwa kofuzo lwesiphawuli kunye nokuhanjiswa kwendawo:
Uhlalutyo lwamaqela
Indibaniselwano yedatha esuka kumaqela omabini kunye neqela kwakhona:
Iijini zokumakisha zamaqela amatsha:
Phonononga inkqubela phambili eququzelelwa ziinkonzo ze-BMKGene zokukhuphela ngetekhnoloji ye-BMKManu S1000 kolu papasho lubonakalisiweyo:
Ingoma, X. et al. (2023) 'I-Spatial transcriptomics ityhila iiseli ze-chlorenchyma ezenziwe ngokukhanya ezibandakanyekayo ekukhuthazeni ukuvuselelwa kokudubula kwi-tomato callus',Iinkqubo zeZiko leSizwe leSayensi lase-United States of America, 120(38), iphe. e2310163120. doi: 10.1073/pnas.2310163120
Wena, Y. et al. (2023) 'Uthelekiso olucwangcisiweyo lolandelelwano-based spatial transcriptomic method',i-bioRxiv, iphe. 2023.12.03.569744. doi: 10.1101/2023.12.03.569744.
