BMKMANU S1000 Spatial Transcriptome
BMKMANU S1000 Spatial Transcriptome Technical Scheme
Features
● Kugadziriswa: 5 µM
● Spot Diameter: 2.5 µM
● Nhamba yenzvimbo: anenge Mamiriyoni maviri
● 3 anogona kutora nzvimbo mafomati: 6.8 mm * 6.8 mm, 11 mm * 11 mm kana 15 mm * 20 mm
● Bead yega yega ine barcode yakatakurwa neprimers inoumbwa nezvikamu zvina:
poly(dT) muswe wemRNA priming uye cDNA synthesis
Unique Molecular Identifier (UMI) kugadzirisa kusarerekera kweamplification
Spatial barcode
Kusunga kutevedzana kwechidimbu kuverenga 1 kutevedzana kwekutanga
● H & E uye kusvibiswa kwefluorescent kwezvikamu
● Kugona kushandisacell segmentation tekinoroji: kubatanidzwa kweH & E staining, fluorescent staining, uye RNA sequencing kuti uone miganhu yesero rega rega uye nenzira kwayo kugovera gene expression kune yega yega.
Zvakanakira zveBMKMANU S1000
●Sub-cellular Resolution: Nzvimbo yega yega yekubata ine > 2 miriyoni spatial Barcoded Spots ine dhayamita ye 2.5 µm uye kupatsanurwa kwe5 µm pakati penzvimbo dzemavara, zvichiita kuti spatial transcriptome kuongororwa ine sub-cellular resolution (5 µm).
●Multi-level Resolution Analysis:Flexible multi-level ongororo kubva ku100 μm kusvika 5 μm kugadzirisa akasiyana matishu maficha pakugadziriswa kwakaringana.
●Kugona Kushandisa "Vatatu mune imwe siraidhi" Cell Segmentation Technology:Kubatanidza fluorescence staining, H&E staining, uye RNA kutevedzana pane imwechete siraidhi, yedu "matatu-mu-imwe" yekuongorora algorithm inosimbisa kuzivikanwa kwesero miganhu yeanotevera cell-based transcriptomics.
●Inoenderana neMazhinji Sequencing Platform: Dzese NGS uye yakareba-yekuverenga kutevedzana iripo.
●Flexible Dhizaini ye1-8 Active Capture Area: Saizi yenzvimbo yekutora inochinjika, zvichikwanisika kushandisa 3 mafomati (6.8 mm * 6.8 mm., 11 mm * 11 mm uye 15 mm * 20 mm)
●One-stop Service: Inosanganisa ruzivo rwese uye hunyanzvi-hwakavakirwa matanho, kusanganisira cryo-chikamu, kusvibisa, tishu optimization, spatial barcoding, kugadzirira raibhurari, kutevedzana, uye bioinformatics.
●Yakakwana Bioinformatics uye Mushandisi-inoshamwaridzika Kuona Kwemhedzisiro:Pasuru inosanganisira makumi maviri nemapfumbamwe ekuongorora uye 100+ yemhando yepamusoro nhamba, yakasanganiswa nekushandiswa kweiyo inhouse yakagadziridzwa software yekuona uye kugadzirisa kupatsanurwa kwesero uye kubatanidza nzvimbo.
●Customized Data Analysis uye Visualization: inowanikwa kune zvakasiyana zvikumbiro zvekutsvaga
●Yakanyanya-unyanzvi Technical Team: ine ruzivo mumhando dzinopfuura mazana maviri nemakumi mashanu emhando dzematishu uye 100+ mhando dzinosanganisira vanhu, mbeva, zvinoyamwisa, hove uye zvidyarwa.
●Real-time Updates pane Yese Project: nekutonga kwakazara kwekufambira mberi kwekuedza.
●Optional Joint Ongororo ine Imwechete-sero mRNA Sequencing
Sevhisi Zvinotsanangurwa
|
Muenzaniso Zvinodiwa
| Library |
Sequencing strategy
| Data inokurudzirwa | Quality Control |
| OCT-yakamisikidzwa cryo samples, 3 zvidhinha pamuenzaniso | S1000 cDNA raibhurari | Illumina PE150 (mamwe mapuratifomu aripo) | 100K PE inoverenga pa100 M ( 60-150 Gb ) | RIN>7 |
Kuti uwane rumwe ruzivo nezve nhungamiro yegadziriro yemuenzaniso uye mafambiro ebasa, ndapota inzwa wakasununguka kutaura kune aBMKGENE nyanzvi
Kuyerera Kwebasa Rebasa
Muchikamu chekugadzirira chemuenzaniso, yekutanga kuwanda kweRNA kutorwa kuyedzwa kunoitwa kuti ive nechokwadi chekuti RNA yemhando yepamusoro inogona kuwanikwa. Muchikamu chekugadzirisa matishu zvikamu zvakasvibiswa uye zvinoonekwa uye mamiriro epermeabilization ekuburitswa kwemRNA kubva munyama anogadziriswa. Iyo yakagadziridzwa protocol inozoshandiswa panguva yekuvaka raibhurari, ichiteverwa nekutevedzana uye kuongororwa kwedata.
Iyo yakazara sevhisi kufambiswa kwebasa kunosanganisira chaiyo-nguva yekuvandudza uye kusimbiswa kwevatengi kuchengetedza inopindura mhinduro loop, kuve nechokwadi chekuita kwakatsetseka kweprojekiti.
Iyo data yakagadzirwa neBMKMANU S1000 inoongororwa uchishandisa software "BSTMatrix", iyo yakazvimiririra yakagadzirwa neBMKGENE, inogadzira Gene Expression Matrix. Kubva ipapo, mushumo wakajairika unogadzirwa unosanganisira kutonga kwemhando yedata, kuongorora kwemukati-sample uye kuongororwa kweboka.
● Kudzora Hunhu hweData:
Kubuda kwedata uye kugovera kwemhando yepamusoro
Kuonekwa kwemajini pane imwe nzvimbo
Tissue coverage
● Ongororo yemukati-yemuenzaniso:
Gene kupfuma
Spot clustering, kusanganisira yakaderedzwa dimension analysis
Differential expression analysis pakati pemasumbu: identification of marker genes
Inoshanda annotation uye hupfumi hwemaka gene
● Ongororo yemapoka:
Kubatanidzwazve kwemavara kubva kune ese masampuli (eg. ane chirwere uye kudzora) uye kubatanidza zvakare
Kuzivikanwa kwemakaki genes kune yega yega cluster
Inoshanda annotation uye hupfumi hwemaka gene
Differential Kuratidzwa kwechikwata chimwechete pakati pemapoka
Pamusoro pezvo, BMKGENE yakagadziridzwa "BSTViewer" chishandiso chine mushandisi-chinogonesa mushandisi kuona mataurirwo ejini uye kuungana kwemavara pazvisarudzo zvakasiyana.
BMKGene yakagadzira software yemushandisi ane hushamwari kuona
BSTViewer nzvimbo inoungana pa-multi-level resolution
BSTCellViewer: otomatiki uye manual cell kupatsanura
Inner-sample Analysis
Spot clustering:
Kuzivikanwa kwemajini emakaki uye kugovera kwenzvimbo:
Inter-group Analysis
Musanganiswa wedata kubva kumapoka maviri uye re-cluster:
Marker genes emasumbu matsva:
Ongorora kufambira mberi kwakafambiswa neBMKGene's spatial transcriptomics masevhisi neiyo BMKManu S1000 tekinoroji mune ino chinyorwa chinyorwa:
Rwiyo, X. et al. (2023) 'Spatial transcriptomics inoratidza chiedza-induced chlorenchyma masero anobatanidzwa mukusimudzira kutsva kwepfuti mumatomato callus',Nyaya dzeNational Academy of Sciences yeUnited States of America, 120(38), p. e2310163120. doi: 10.1073/pnas.2310163120
Iwe, Y. et al. (2023) 'Systematic kuenzanisa yekuenzanisa-yakavakirwa spatial transcriptomic nzira',bioRxiv, p. 2023.12.03.569744. doi: 10.1101/2023.12.03.569744.
