Detectio ion of f miRN iRNA by Activ tive Ca Capture + Dig igit ital l Co Countin ing (A (AC+ C+DC) ) Micr icrosc scopy Taylor Canady Carl R. Woese Fellow @ UIUC 1
Liq iquid id Bio iopsy: Bio ioflu luid id Bio iomarker r Detectio ion Op Opportunity of of Liq Liquid Bi Biopsy • Non-invasive • Early detection & quantification • Sort mutation types • Fast monitor and improve treatment Challe Cha lenges of of Liq Liquid Bi Biopsy • Targets Disease Patient Diversity • Sequence similarity (SNPs) • Concentration & matrix • Build POC assay compatibility (time, cost, ...) Des Desig ign Co Constr traints • Broad single base selectivity (DNA Nano) • Extreme sensitivity (PC Biosensors) • Wide dynamic range • No amp, no wash • Assay time: minutes to few hours 2 Clin Cancer Res; 23(10) 2017
Engin ineeri ring Tools ls for r li liquid id Bio iopsy of miRN iRNA Bio iomarkers Photonic ic Cry Crystal l Bio Biosensors DNA Nanotechnolo logy 3
Prin rincip iple le of f Photonic ic Cry Crystal l Nan anopartic icle le Se Sensin ing Computer simulation: Finite-Difference Time-Domain (FDTD) Evanescent Field 400 nm |E| 2 (v 2 /m 2 ) θ Δ I (PIV) Reflection (A.U.) λ Δ λ (PWV) . 625 Wavelength (nm) Peak Wavelength Value ( PWV ) Measure Peak Intensity Value ( PIV )
Digital Resolution of AuNP “Tags”: Detection Method Hybrid causes drop in PC reflectance peak intensity & shift in Allows for digital resolution of particles 400 nm wavelength 25 µm 5 Canady et al. PNAS, 2019
Activ tive Ca Capture + Dig igit ital l Co Countin ing (A (AC+ C+DC) ) miRN iRNA Dia iagnostic ics 6 Canady et al. PNAS, 2019
2 hour endpoint miR-375 + AuNPs Canady et al. PNAS, 2019 Background 0.1 fM 1 fM 10 fM 100 fM 1 pM 10 pM 1 min 30 mins miR-375 + AuNPs 60 mins 90 mins 120 mins 7 625.0 625.3 25 µm
Highlights ld @ resonance nan Photonic Crystal Bios iosensin ing: Enhanced E-fie ield anoparticle detection by ∆G RXN ≅ 0 si Toe oehold ld DN DNA Probes: Optimize hyb ybrid idization by single le base ase se sele lectiv ivity AC+DC Ass ssay: : Co Combin ine AuNP-DNA probe (s (selec lectiv ivit ity) ) & PC C bio iose sensors (s (sensit itiv ive) ) for r rapid id, no ampli lifi ficatio ion, and no wash miRN iRNA detectio ion AC+DC Data: ith 10 6 dynamic miRN iRNA detectio ion with ic range (1 (100 aM aM lo lowest) Sin Single le base se selec lectiv ivit ity across 22mer miRN iRNA (5 (5 example les) Detectio ion in in compli licated background (excess non-target RNA) Futu ture: Micr icroflu fluid idic ic im impla lantatio ion, addit itio ional l photonic ic-mic icroscopes 8
Recommend
More recommend
