BMKMANU S1000 Spatial Transcriptome
BMKMANU S1000 Spatial Transcriptome Technical Scheme
Likaroloana
● Qeto: 5 µM
● Spot Diameter: 2.5 µM
● Palo ea libaka: hoo e ka bang limilione tse 2
● 3 lifomate tse ka khonehang tsa sebaka sa ho hapa: 6.8 mm * 6.8 mm, 11 mm * 11 mm kapa 15 mm * 20 mm
● Sefaha se seng le se seng se nang le barcode se kentsoe li-primers tse entsoeng ka likarolo tse 4:
poly(dT) mohatla oa mRNA priming le cDNA synthesis
Unique Molecular Identifier (UMI) ho lokisa leeme la ho holisa
Sebaka sa barcode
Tatelano e tlamang ya karolo e itseng e baletsweng 1 ya tatellano
● H&E le litšila tsa fluorescent tsa likarolo
● Monyetla oa ho sebelisatheknoloji ea ho arola lisele: ho kopanngoa ha litšila tsa H&E, litšila tsa fluorescent, le tatellano ea RNA ho fumana meeli ea sele e 'ngoe le e' ngoe le ho abela ka nepo polelo ea lefutso ho sele ka 'ngoe.
Melemo ea BMKMANU S1000
●Sub-cellular Resolution: Sebaka se seng le se seng se nkiloeng se na le > Li-Barcoded Spots tse limilione tse 2 tse bophara ba 2.5 µm le sebaka sa 5 µm lipakeng tsa litsi tsa libaka, se nolofalletsang tlhahlobo ea transcriptome ea sebaka ka qeto ea sub-cellular (5 µm).
●Tlhahlobo ea Liqeto tse ngata:Tlhahlobo e feto-fetohang ea maemo a mangata ho tloha ho 100 μm ho isa ho 5 μm ho rarolla likarolo tse fapaneng tsa lisele ka tharollo e nepahetseng.
●Monyetla oa ho Sebelisa "Tse tharo selaeteng se le seng" Theknoloji ea Karolo ea Cell:Ho kopanya mabala a fluorescence, H&E staining, le tatellano ea RNA selaeteng se le seng, algorithm ea rona ea tlhahlobo ea "tharo-in-one" e matlafatsa ho tsebahatsa meeli ea lisele bakeng sa li-transcriptomics tse latelang tse thehiloeng liseleng.
●E lumellana le Lipolanete tse ngata tsa Tatelano: Ka bobeli NGS le tatellano ea nako e telele ea ho bala e fumaneha.
●Moralo o Fetohang oa 1-8 Active Capture Area: Boholo ba sebaka sa ho hapa sea fetoha, ho khoneha ho sebelisa liforomo tse 3 (6.8 mm * 6.8 mm., 11 mm * 11 mm le 15 mm * 20 mm)
●Tšebeletso e le 'ngoe: E kopanya boiphihlelo bohle le mehato e ipapisitseng le boiphihlelo, ho kenyelletsa le cryo-sectioning, staining, optimization ea lisele, barcoding ea sebaka, ho lokisa laeborari, tatellano, le bioinformatics.
●Bioinformatics e Felletseng le Ponahalo ea Liphetho e bonolo ho basebelisi:sephutheloana akarelletsa 29 analysis le 100+ boleng bo phahameng lipalo, ho kopantswe le tshebediso ya inhouse ntshetswa pele Software ho bona ka mahlo a kelello le Customize sele petsoha le letheba clustering.
●Tshekatsheko ya Ditaba tse Ikgethehileng le Pono: e fumaneha bakeng sa likopo tse fapaneng tsa lipatlisiso
●Sehlopha sa Tekheniki se nang le litsebo tse phahameng: o na le boiphihlelo mefuteng ea linama tse fetang 250 le mefuta e fetang 100+ ho kenyeletsoa batho, litoeba, liphoofolo tse anyesang, litlhapi le limela.
●Lintlafatso tsa nako ea 'nete morerong oohle: ka taolo e felletseng ea tsoelopele ea liteko.
●Tlhahlobo e Kopanetsoeng ka Boikhethelo ka Tatelano ea sele e le 'ngoe ea mRNA
Litlhaloso tsa Tšebeletso
|
Mohlala Litlhoko
| Laebrari |
Leano la tatellano
| Lintlha li khothalelitsoe | Taolo ea Boleng |
| Mehlala ea cryo e kentsoeng ka OCT, li-blocks tse 3 ka mohlala | S1000 cDNA library | Illumina PE150 (li-platform tse ling lia fumaneha) | 100K PE e baloa ho 100 uM ( 60-150 Gb ) | RIN>7 |
Bakeng sa lintlha tse ling mabapi le tataiso ea boitokisetso ea mohlala le ts'ebetso ea ts'ebeletso, ka kopo ikutloe u lokolohile ho bua le aBMKGENE setsebi
Phallo ea Mosebetsi oa Ts'ebeletso
Mokhahlelong oa boitokiso oa sampole, teko ea pele ea ho ntša RNA ka bongata e etsoa ho netefatsa hore RNA ea boleng bo holimo e ka fumanoa. Mokhahlelong oa ntlafatso ea lithane likarolo li silafatsoa le ho bonoa, 'me maemo a permeabilization bakeng sa tokollo ea mRNA ho tsoa liseleng a ntlafatsoa. Protocol e ntlafalitsoeng e sebelisoa nakong ea kaho ea laeborari, e lateloe ke tatellano le tlhahlobo ea data.
Ts'ebetso e felletseng ea ts'ebeletso e kenyelletsa lintlafatso tsa nako ea nnete le netefatso ea bareki ho boloka sephetho se arabelang, ho netefatsa ts'ebetso ea projeke e bonolo.
Lintlha tse hlahisoang ke BMKMANU S1000 li hlahlojoa ho sebelisoa software "BSTMatrix", e ikemetseng e entsoeng ke BMKGENE, e hlahisang Gene Expression Matrix. Ho tloha moo, ho hlahisoa tlaleho e tloaelehileng e kenyeletsang taolo ea boleng ba data, tlhahlobo ea ka hare-sample le tlhahlobo ea lihlopha tse fapaneng.
● Taolo ea Boleng ba Boitsebiso:
Tlhahiso ea lintlha le kabo ea lintlha tsa boleng
Ho fumanoa ha liphatsa tsa lefutso sebakeng ka seng
Ho koaheloa ha lisele
● Tlhahlobo ea mohlala oa ka hare:
Leruo la liphatsa tsa lefutso
Spot clustering, ho kenyeletsoa tlhahlobo e fokolitsoeng ea litekanyo
Tlhahlobo e fapaneng ea polelo lipakeng tsa lihlopha: ho tsebahatsa liphatsa tsa lefutso tsa matšoao
Tlhaloso e sebetsang le ntlafatso ea liphatsa tsa lefutso tsa matšoao
● Tlhahlobo ea lihlopha tse fapaneng:
Ho kopanngoa bocha ha matheba ho tloha lisampoleng ka bobeli (mohlala, ho kula le ho laola) le ho kopanya hape
Khethollo ea liphatsa tsa lefutso tsa sehlopha ka seng
Tlhaloso e sebetsang le ntlafatso ea liphatsa tsa lefutso tsa matšoao
Phapang Polelo ea sehlopha se tšoanang lipakeng tsa lihlopha
Ho feta moo, BMKGENE e ntlafalitse "BSTViewer" ke sesebelisoa se bonolo sa mosebelisi se nolofalletsang mosebelisi ho bona ka mahlo a kelello polelo ea lefutso le ho bokellana ha lihlopha liqetong tse fapaneng.
BMKGene e hlahisitse software bakeng sa pono e bonolo ea basebelisi
Sehlopha sa BSTViewer se kopantsoe ka qeto ea maemo a mangata
BSTCellViewer: karohano ea lisele tsa othomathike le ka letsoho
Inner-sample Analysis
Spot clustering:
Boitsebiso ba liphatsa tsa lefutso le kabo ea sebaka:
Inter-group Analysis
Motsoako oa data ho tsoa lihlopheng ka bobeli le ho kopanya hape:
Liphatsa tsa lefutso tsa lihlopha tse ncha:
Lekola tsoelopele e tsamaisoang ke lits'ebeletso tsa transcriptomics tsa BMKGene ka theknoloji ea BMKManu S1000 khatisong ena e hlahang:
Sefela, X. et al. (2023) 'Spatial transcriptomics e senola lisele tsa chlorenchyma tse hlahisitsoeng ke leseli tse amehang ho khothaletse nchafatso ea letlobo ka har'a tomate callus',Ts'ebetso ea National Academy of Sciences ea United States of America, 120(38), leq. e2310163120. doi: 10.1073/pnas.2310163120
Uena, Y. le ba bang. (2023) 'Papiso e hlophisitsoeng ea mekhoa ea tatellano e thehiloeng ho sebaka sa transcriptomic',bioRxiv, leq. 2023.12.03.569744. doi: 10.1101/2023.12.03.569744.
