10x Genomics Visium Spatial Transcriptome
Nā hiʻohiʻona
● Hoʻoholo: 100 µM
● Anawaena kiko: 55 µM
● Ka helu o nā kiko: 4992
● Wahi paʻa: 6.5 x 6.5 mm
● Hoʻopiha ʻia kēlā me kēia wahi barcode me nā mea mua i haku ʻia me 4 mau ʻāpana:
- poly(dT) huelo no ka mRNA priming a me cDNA synthesis
- Unique Molecular Identifier (UMI) no ka hoʻoponopono ʻana i ka manaʻo hoʻonui
- Kāleka pāʻālua
- Ke kaʻina hoʻopaʻa ʻana o ka helu helu ʻāpana 1 sequencing primer
● H&E hoʻomaʻemaʻe i nā ʻāpana
Pono
●ʻO ka lawelawe hoʻokahi: hoʻohui i nā ʻike āpau a me nā ʻanuʻu e pili ana i ke akamai, me ka cryo-sectioning, staining, tissue optimization, spatial barcoding, hoʻomākaukau hale waihona puke, sequencing a me bioinformatics.
● Kiʻekiʻe akamai kime loea: me ka ʻike ma luna o 250 mau ʻano kiko a me 100+ mau ʻano me ke kanaka, ka ʻiole, ka mammal, ka iʻa a me nā mea kanu.
●Hoʻopuka manawa maoli i ka papahana holoʻokoʻa: me ka mana piha o ka holomua hoʻokolohua.
●Nā bioinformatic maʻamau piha:Aia i loko o ka pūʻolo he 29 kānana a me 100+ kiʻi kiʻekiʻe.
●ʻIkepili ʻikepili a me ka nānā ʻana: loaʻa no nā noi noiʻi like ʻole.
●ʻO ka hoʻopaʻa ʻana hui pū me ka hoʻopaʻa ʻana o ka mRNA pū hoʻokahi
Nā kikoʻī
| Nā Koina Laʻana | Hale Waihona Puke | Hoʻolālā kaʻina | Manaʻo ʻia ka ʻikepili | Ka Mana Mana |
| Nā laʻana cryo i hoʻokomo ʻia i ka OCT, nā laʻana FFPE (Ka anawaena oi loa: ma kahi o 6x6x6 mm3) 3 poloka no ka la'ana | 10X Visium cDNA waihona | Illumina PE150 | 50K PE heluhelu no kēlā me kēia wahi (60Gb) | RIN>7 |
No nā kikoʻī hou aku e pili ana i ke alakaʻi hoʻomākaukau laʻana a me ke kahe hana lawelawe, e ʻoluʻolu e kamaʻilio me aloea BMKGENE
Hana Hana Hana
Ma ka papa hoʻomākaukau laʻana, hana ʻia kahi hoʻāʻo hoʻāʻo RNA hoʻāʻo mua e hōʻoia i hiki ke loaʻa kahi RNA kiʻekiʻe.Ma ke kahua hoʻomaʻamaʻa kikoʻī ua ʻike ʻia nā ʻāpana a ʻike ʻia nā kūlana permeabilization no ka hoʻokuʻu ʻana o ka mRNA mai ka ʻiʻo.Hoʻohana ʻia ka protocol i hoʻopaʻa ʻia i ka wā o ke kūkulu ʻana i ka hale waihona puke, a ukali ʻia e ka sequencing a me ka nānā ʻana i ka ʻikepili.
Loaʻa ka holo kaʻa hana piha i ka manawa maoli a me ka hōʻoia ʻana o ka mea kūʻai aku e mālama i kahi loop pane pane, e hōʻoia i ka hoʻokō ʻana o ka papahana.
Loaʻa i kēia ka nānā ʻana:
Ka Mana Pono ʻIkepili:
ʻO ka hoʻopuka ʻikepili a me ka hāʻawi ʻana i ka helu maikaʻi
ʻO ka ʻike ʻana i nā Gene ma kēlā me kēia wahi
o Ka uhi ʻana o ke kino
Ka nānā ʻana i loko o ka laʻana:
o Ka waiwai Gene
ʻO ka hui ʻana o nā kiko, me ka hoʻoemi ʻana i ke ana
o Ka nānā 'ana i ka 'ōlelo 'oko'a ma waena o nā pū'ulu: 'ike 'ana i nā genes marker
ʻO ka hōʻike kikoʻī a me ka hoʻonui ʻana i nā genes marker
ʻIkepili waena
o Ka hui hou ʻana o nā kiko mai nā laʻana ʻelua (e like me ka maʻi a me ka hoʻomalu) a me ka hui hou ʻana
o Ka ʻike ʻana i nā genes marker no kēlā me kēia pūʻulu
ʻO ka hōʻike kikoʻī a me ka hoʻonui ʻana i nā genes marker
o Hōʻike ʻokoʻa o ka pūʻulu like ma waena o nā pūʻulu
Hoʻolāʻau i loko
Hoʻopili kiko
ʻO ka ʻike ʻana i nā genes marker a me ka hoʻohele ākea
ʻIkepili waena
Hoʻohui ʻikepili mai nā hui ʻelua a me ka hui hou
Nā genes marker o nā pūʻulu hou
E ʻimi i nā holomua i hoʻomaʻamaʻa ʻia e ka lawelawe spatial transcriptomics o BMKGene e 10X Visium Ma kēia mau puke i hōʻike ʻia:
Chen, D. et al.(2023) 'mthl1, kahi Drosophila homologue o mammalian adhesion GPCRs, ua komo i loko o ka antitumor reactions i injected oncogenic mau nalo', Proceedings of the National Academy of Sciences of the United States of America, 120(30), p.e2303462120.doi: /10.1073/pnas.2303462120
Chen, Y. et al.(2023) 'O STEEL ka mea e hiki ai i ka ho'ākāka 'ana i ka spatiotemporal transcriptomic data', Briefings in Bioinformatics, 24(2), pp. 1–10.doi: 10.1093/BIB/BBAD068.
Liu, C. et al.(2022) 'A spatiotemporal atlas of organogenesis in the development of orchid flowers', Nucleic Acids Research, 50(17), pp. 9724–9737.doi: 10.1093/NAR/GKAC773.
Wang, J. et al.(2023) 'Integrating Spatial Transcriptomics and Single-nucleus RNA Sequencing Reveals the Potential Therapeutic Strategies for Uterine Leiomyoma', International Journal of Biological Sciences, 19(8), pp. 2515–2530.doi: 10.7150/IJBS.83510.
