BMKMANU S3000_Spatial Transcriptome
Tsarin fasaha na BMKMANU S3000 Spatial Transcriptome
Siffofin
Ƙaddamarwa: 3.5 µM
- Diamita na Tabo: 2.5 µM
- Yawan tabo: kusan Miliyan 4
- Tsarukan yanki mai yuwuwa 3: 6.8mm * 6.8mm, 11mm * 11mm ko 15mm * 20mm
- Kowane bead ɗin da aka ɗora ana ɗora shi da abubuwan da suka ƙunshi sassa 4:
• wutsiya poly(dT) don ƙaddamar da mRNA da haɗin cDNA,
• Mai Gano Kwayoyin Halitta na Musamman (UMI) don gyara son zuciya na haɓakawa
• Barcode na sarari
Jerin dauri na juzu'i na karanta juzu'i 1 na farko
- H&E da tabon sassa
- Yiwuwar yin amfani da fasahar rabuwar tantanin halitta: haɗakar da tabon H&E, tabo mai kyalli, da jerin RNA don ƙayyade iyakokin kowane tantanin halitta kuma daidai sanya bayanin kwayar halitta ga kowane tantanin halitta. Ana aiwatar da bincike na bayanan sararin samaniya bisa tushen tantanin halitta.
- Yiwuwa don cimma ƙididdigar ƙuduri mai yawa: Binciken matakai masu sassauƙa da yawa daga 100um zuwa 3.5 um don warware fasalulluka iri-iri a mafi kyawun ƙuduri.
Amfanin BMKMANU S3000
-Ninki biyu na wuraren kamawa zuwa miliyan 4: tare da ingantaccen ƙuduri na 3.5 uM, wanda ke haifar da mafi girma a cikin kwayar halitta da gano UMI a kowane tantanin halitta. Wannan yana haifar da ingantattun tarin sel dangane da bayanan rubutu, tare da mafi kyawun daki-daki wanda ya dace da tsarin kyallen takarda.
- Ƙaddamar da ƙananan salula:Kowane wurin da aka kama yana ƙunshe da> Barcoded Spots miliyan 2 tare da diamita na 2.5 µm da tazara na 5 µm tsakanin wuraren tabo, yana ba da damar nazarin bayanan sararin samaniya tare da ƙudurin ƙaramin salula (5 μm).
-Binciken ƙuduri mai yawa:Binciken matakai masu sassaucin ra'ayi daga 100 μm zuwa 5 μm don warware fasalin nama iri-iri a mafi kyawun ƙuduri.
-Yiwuwar amfani da fasahar rabuwar tantanin halitta "Uku cikin zamewa ɗaya:haɗe da tabon haske, tabon H&E, da jerin RNA akan faifai guda ɗaya, binciken mu na "uku-in-daya" algorithm yana ba da ikon gano iyakokin tantanin halitta don bayanan tushen tantanin halitta na gaba.
-Mai jituwa tare da dandamali masu yawa na jeri: duka NGS da jerin dogon karantawa akwai.
-Zane mai sassauƙa na yanki 1-8 mai aiki: girman wurin kama yana da sassauƙa, yana yiwuwa a yi amfani da tsarin 3 (6.8mm * 6.8 mm., 11 mm * 11 mm and 15 mm * 20 mm)
-Sabis na tsayawa ɗaya: ya haɗu da duk ƙwarewa da matakai na tushen fasaha, ciki har da sassan cryo-sectioning, tabo, inganta kyallen takarda, ƙirar sararin samaniya, shirye-shiryen ɗakin karatu, sequencing, da bioinformatics.
-Cikakken bioinformatics da hangen nesa mai amfani na sakamako:kunshin ya ƙunshi nazari 29 da ƙididdiga masu inganci 100+, haɗe tare da yin amfani da ingantaccen software na cikin gida don hangowa da tsara rarrabuwar tantanin halitta da tari.
-Kirkirar bayanai da hangen nesa: akwai don buƙatun bincike daban-daban
-Ƙwararrun ƙwararrun ƙungiyar fasaha: tare da gogewa a cikin nau'ikan nama sama da 250 da nau'ikan 100+ da suka haɗa da ɗan adam, linzamin kwamfuta, dabbobi masu shayarwa, kifi da tsirrai.
-Sabuntawa na ainihi akan gaba dayan aikin: tare da cikakken iko na ci gaban gwaji.
-Binciken haɗin gwiwa na zaɓi tare da jeri na mRNA guda-cell
Ƙayyadaddun Sabis
| Samfuran Bukatun | Laburare | Dabarun Jeri | An Shawarar Bayanai | Kula da inganci |
| Samfuran cryo masu kunshe da OCT (Mafi kyawun diamita: kusan. 6×6×6mm³) 2 tubalan kowane samfurin 1 don gwaji, 1 don madadin | S3000 cDNA Library | Farashin PE150 | 160K PE ana karantawa akan 100 υM (250 Gb) | RIN> 7 |
Don ƙarin cikakkun bayanai kan jagorar shirye-shiryen samfuri da aikin sabis, da fatan za a ji daɗin magana da aMasanin BMKGENE
Gudun Aikin Sabis
A cikin lokacin shirye-shiryen samfurin, ana yin gwajin haƙar RNA na farko don tabbatar da samun RNA mai inganci. A cikin matakin inganta nama sassan sassan suna da tabo kuma an duba su kuma an inganta yanayin ƙetare don sakin mRNA daga nama. Sannan ana amfani da ingantacciyar ƙa'idar yayin aikin ginin ɗakin karatu, sannan a bi da bi da kuma nazarin bayanai.
Cikakken aikin sabis ɗin ya haɗa da sabuntawa na ainihi da kuma tabbatar da abokin ciniki don kula da madaidaicin ra'ayi, yana tabbatar da aiwatar da aikin mai santsi.
An yi nazarin bayanan da BMKMANU S3000 ya ƙirƙira ta amfani da software na "BSTMatrix", wanda BMKGENE ya tsara shi da kansa, yana samar da matakin tantanin halitta da ƙuduri mai yawa Gene Expression Matrix. Daga can, ana samar da daidaitaccen rahoto wanda ya haɗa da sarrafa ingancin bayanai, bincike-bincike na ciki da bincike tsakanin ƙungiyoyi.
- Kula da ingancin bayanai:
- Fitowar bayanai da rarraba makin inganci
- Gano kwayoyin halitta a kowane tabo
- Ciwon nama
- Binciken samfurin ciki:
- Halin arziki
- Tarin tabo, gami da raguwar ƙididdigar girma
- Bambance-bambancen maganganun magana tsakanin gungu: gano kwayoyin halitta masu alama
- Bayanin aiki da haɓaka ƙwayoyin halitta masu alama
- Bincike tsakanin ƙungiyoyi
- Sake haɗa tabo daga samfuran duka biyu (misali marasa lafiya da sarrafawa) da sake tari
- Gano kwayoyin halitta masu alama ga kowane gungu
- Bayanin aiki da haɓaka ƙwayoyin halitta masu alama
- Bambance-banbance na gungu iri ɗaya tsakanin ƙungiyoyi
Bugu da ƙari, BMKGene ya ɓullo da "BSTViewer" kayan aiki ne na abokantaka mai amfani wanda ke ba mai amfani damar hango maganganun kwayoyin halitta da tabo tari a matakai daban-daban.
BMKGene yana ba da sabis na bayanan sararin samaniya akan madaidaicin ƙudurin tantanin halitta guda ɗaya (dangane da bin tantanin halitta ko murabba'in murabba'i daga 100um zuwa 3.5um).
Bayanan bayanan sararin samaniya daga sassan nama akan faifan S3000 an yi su da kyau kamar ƙasa.
Nazarin shari'a 1: Kwakwalwar linzamin kwamfuta
Binciken sashin kwakwalwar linzamin kwamfuta tare da S3000 ya haifar da gano ~ 94 000 Kwayoyin, tare da tsaka-tsaki jerin ~ 2000 genes ta tantanin halitta. Ingantacciyar ƙuduri na 3.5 uM ya haifar da cikakken tari na sel dangane da tsarin rubutun, tare da gungu na sel waɗanda ke kwaikwayi nau'ikan bambance-bambancen tsarin kwakwalwa. Ana iya lura da wannan cikin sauƙi ta hanyar hangen nesa da rarraba ƙwayoyin sel waɗanda aka taru azaman oligodendrocytes da ƙwayoyin microglia, waɗanda kusan keɓaɓɓu keɓaɓɓu a cikin launin toka da fari, bi da bi.
Nazarin shari'a 2: Mouse amfrayo
Binciken sashin amfrayo na linzamin kwamfuta tare da S3000 ya haifar da gano ~ 2200 000 Kwayoyin, tare da tsaka-tsakin jeri na ~ 1600 genes a kowace tantanin halitta. Ingantacciyar ƙuduri na 3.5 uM ya haifar da cikakken tarin ƙwayoyin sel dangane da tsarin rubutu, tare da gungu 12 a cikin yanki na ido da kuma gungu 28 a cikin yanki na kwakwalwa.
Ciki-samfurin Tarin Tarin Kwayoyin Halitta:
Gane kwayoyin halitta masu alamar alama da rarraba sarari:
- ƙuduri mafi girma: Idan aka kwatanta da S1000 slide, kowane yanki na S3000 yana ƙunshe da> 4 miliyan Barcoded Spots na sararin samaniya tare da diamita na 2.5 µm da nisa na 3.5 µm tsakanin wuraren tabo, yana ba da damar bincike na sararin samaniya tare da ƙudurin sub-cellular (square bin: 3.5 µm).
- Ingantacciyar kamawa: idan aka kwatanta da S1000 slide, Median_UMI ya karu daga 30% zuwa 70%, Median_Gene ya karu daga 30% zuwa 60%
Tsarin guntu na S1000:
Tsarin guntu S3000:
