BMKMANU S3000_Spatial transcriptome
Atụmatụ nka nka BMKMANU S3000 Spatial Transcriptome
Atụmatụ
- Mkpebi: 3.5 µM
- Dayameta ntụpọ: 2.5 µM
- Ọnụọgụ ntụpọ: ihe dịka nde anọ
- 3 enwere ike ijide mpaghara mpaghara: 6.8 mm * 6.8 mm, 11 mm * 11 mm ma ọ bụ 15 mm * 20 mm
- A na-eju bead ọ bụla etinyere na primers nke nwere ngalaba 4:
• poly(dT) ọdụ maka mRNA priming na cDNA njikọ,
• Identifier Molecular Pụrụ Iche (UMI) iji dozie nhụsianya mmụba
• Koodu oghere
• Usoro njide nke akụkụ agụ 1 usoro primer
- H&E na fluorescent ntụpọ nke ngalaba
- Ohere iji teknụzụ nkewa cell: njikọta nke ntụpọ H&E, ntụpọ fluorescent, na usoro RNA iji chọpụta ókè sel ọ bụla wee kenye mkpụrụ ndụ mkpụrụ ndụ n'ụzọ ziri ezi na sel ọ bụla. Na-ahazi nyocha profaịlụ mbara ala nke mgbada dabere na ọnụọgụ cell.
- Enwere ike nweta nyocha mkpebi multilevel: nyocha ọkwa dị iche iche na-agbanwe agbanwe sitere na 100um ruo 3.5 um iji dozie atụmatụ anụ ahụ dị iche iche na mkpebi kacha mma.
Uru nke BMKMANU S3000
-Eji okpukpu abụọ nke njide tụrụ na nde 4: na mkpebi dị mma nke 3.5 uM, na-eduga na mkpụrụ ndụ dị elu na nchọpụta UMI n'otu sel. Nke a na-ebute mkpokọta mkpụrụ ndụ ka mma dabere na profaịlụ ederede, yana nkọwa kacha mma dabara na nhazi anụ ahụ.
- Mkpebi sub-cellular:Mpaghara njide ọ bụla nwere> 2 nde spatial Barcoded Spots nwere dayameta nke 2.5 µm na oghere nke 5 µm n'etiti ebe ntụpọ, na-enyere nyocha transcriptome ohere na mkpebi sub-cellular (5 µm).
-Nyocha mkpebi ọtụtụ ọkwa:Nyocha ọtụtụ ọkwa na-agbanwe agbanwe sitere na 100 μm ruo 5 μm iji dozie atụmatụ anụ ahụ dị iche iche na mkpebi kacha mma.
-Enwere ike iji teknụzụ nkewa mkpụrụ ndụ "Atọ n'ime otu slide:ijikọta ntụpọ fluorescence, ntụpọ H&E, na usoro RNA n'otu slide, nyocha algorithm anyị "atọ na otu" na-enye ike njirimara nke oke cell maka transcriptomics dabere na cell na-esote.
-Dakọtara na ọtụtụ usoro nhazi usoro: ma NGS na usoro ọgụgụ ogologo dị.
-Nhazi na-agbanwe agbanwe nke mpaghara njide 1-8 na-arụ ọrụ: nha nke mpaghara njide na-agbanwe agbanwe, enwere ike iji usoro 3 (6.8 mm * 6.8 mm., 11 mm * 11 mm na 15 mm * 20 mm)
-Ọrụ nkwụsịtụ: na-ejikọta ahụmịhe niile na usoro dabere na nka, gụnyere ngalaba cryo, ncha, njikarịcha anụ ahụ, oghere oghere, nkwadebe ọba akwụkwọ, usoro, na bioinformatics.
-Bioinformatics zuru oke na nhụhụta enyi-enyi nke onye ọrụ nke nsonaazụ:ngwugwu gụnyere nyocha 29 na ọnụ ọgụgụ dị elu 100+, jikọtara ya na iji ngwa ime ụlọ emepụtara iji hụ anya na hazie nkewa cell na nchịkọta ntụpọ.
-Nyocha na nleba anya data ahaziri: dị maka arịrịọ nyocha dị iche iche
-Otu egwuregwu nka nwere oke nka: nwere ahụmahụ n'ihe karịrị ụdị anụ ahụ 250 na ụdị 100+ gụnyere mmadụ, òké, mammamal, azụ na osisi.
-Mmelite ozugbo na ọrụ niile: na njikwa zuru oke nke ọganihu nnwale.
-Nyocha nkwonkwo nhọrọ na usoro mRNA otu-cell
Nkọwapụta ọrụ
| Ihe atụ chọrọ | Ụlọ akwụkwọ | Usoro usoro | Akwadoro data | Njikwa ogo |
| Nlereanya cryo gbanyere OCT ( Dayameta kacha mma: ihe ruru. 6 × 6 × 6 mm³) 2 ngọngọ kwa sample 1 maka nnwale, 1 maka ndabere | S3000 cDNA Library | Ilumina PE150 | 160K PE na-agụ kwa 100 υM (250 Gb) | RIN> 7 |
Maka nkọwa ndị ọzọ na ntụzịaka nkwadebe nlele na usoro ọrụ ọrụ, biko nweere onwe gị ịgwa aỌkachamara BMKGENE
Usoro ọrụ ọrụ
N'oge nkwadebe nlele, a na-eme nnwale mmịpụta RNA mbụ iji hụ na enwere ike nweta RNA dị elu. N'ọkwa kachasị mma nke anụ ahụ, akụkụ ndị ahụ nwere ntụpọ ma na-ahụ anya ma na-emeziwanye ọnọdụ mRNA maka mwepụta anụ ahụ. A na-etinye usoro ahụ kachasị mma n'oge a na-arụ ụlọ ọba akwụkwọ, na-esote usoro na nyocha data.
Usoro ọrụ ọrụ zuru oke na-agụnye mmelite ozugbo yana nkwenye ndị ahịa iji nọgide na-enwe nzaghachi nzaghachi nzaghachi, na-eme ka mmezu ọrụ dị mma.
A na-enyocha data sitere na BMKMANU S3000 site na iji sọftụwia "BSTMatrix", nke BMKGENE na-emepụta onwe ya, na-emepụta ọkwa cell na mkpebi multilevel Gene Expression Matrix. Site n'ebe ahụ, a na-emepụta akụkọ ọkọlọtọ nke gụnyere njikwa njirimara data, nyocha nke ime na nyocha nke otu.
- Njikwa ogo data:
- Mmepụta data na nkesa akara ogo
- Nchọpụta mkpụrụ ndụ ihe nketa kwa ntụpọ
- Mkpuchi anụ ahụ
- Nleba anya n'ime:
- Ọgaranya Gene
- Nchịkọta ntụpọ, gụnyere nyocha nha dị ala
- Nyocha okwu dị iche iche n'etiti ụyọkọ: njirimara nke mkpụrụ ndụ ihe nrịbama
- Nkọwa ọrụ na ịba ụba nke mkpụrụ ndụ ihe nrịbama
- Nyocha n'etiti otu
- Nchikota nke ntụpọ site na nlele abụọ (dịka ọrịa na njikwa) na nchịkọta ọzọ
- Nchọpụta mkpụrụ ndụ ihe nrịbama maka ụyọkọ ọ bụla
- Nkọwa ọrụ na ịba ụba nke mkpụrụ ndụ ihe nrịbama
- Okwu dị iche iche nke otu ụyọkọ n'etiti otu
Na mgbakwunye, BMKGene mepụtara "BSTViewer" bụ ngwá ọrụ enyi na enyi nke na-enyere onye ọrụ aka iji anya nke uche hụ okwu mkpụrụ ndụ ihe nketa na ntụpọ nchịkọta na mkpebi dị iche iche.
BMKGene na-enye ọrụ profaịlụ gbasara oghere na mkpebi otu cell (dabere na cell bin ma ọ bụ multilevel square bin si 100um ruo 3.5um).
Data profaịlụ gbasara mbara ala sitere na akụkụ anụ ahụ na slide S3000 rụrụ nke ọma dị ka n'okpuru.
Ọmụmụ ikpe 1: ụbụrụ òké
Nyocha nke akụkụ ụbụrụ òké na S3000 rụpụtara na njirimara nke ~ 94 000 sel, yana usoro etiti nke ~ 2000 genes kwa cell. Mkpebi emelitere nke 3.5 uM butere nchịkọta zuru oke nke mkpụrụ ndụ dabere na usoro ederede, yana ụyọkọ mkpụrụ ndụ na-eṅomi ụbụrụ dị iche iche. A na-ahụ nke a n'ụzọ dị mfe site n'ịhụ nkesa nke mkpụrụ ndụ agbakọtara dị ka oligodendrocytes na sel microglia, nke fọrọ nke nta ka ọ bụrụ naanị na isi awọ na ọcha, n'otu n'otu.
Ọmụmụ ikpe 2: Embrayo òké
Nyocha nke akụkụ embrayo òké nwere S3000 rụpụtara na njirimara ~ 2200 000 sel, yana usoro etiti nke ~ 1600 genes kwa cell. Mkpebi emelitere nke 3.5 uM butere nchịkọta zuru oke nke mkpụrụ ndụ dabere na usoro ederede, yana ụyọkọ 12 na mpaghara anya yana ụyọkọ 28 n'akụkụ ụbụrụ.
Ịchịkọta mkpụrụ ndụ nyocha nke ime ime:
Nchọpụta mkpụrụ ndụ ihe nrịbama na nkesa oghere:
- Mkpebi sub-cellular dị elu: E jiri ya tụnyere S1000 slide, mpaghara njide ọ bụla nke S3000 nwere> 4 nde spatial Barcoded Spots na dayameta nke 2.5 µm na anya nke 3.5 µm n'etiti ebe ntụpọ, na-enyere nyocha transcriptome ohere ohere nwere mkpebi sub-cellular dị elu. (square bin: 3.5 µm).
- arụmọrụ dị elu: atụnyere S1000 slide, Median_UMI na-abawanye site na 30% ruo 70%, Median_Gene na-abawanye site na 30% ruo 60%
Atụmatụ nke S1000 mgbawa:
Atụmatụ nke S3000 mgbawa:
