Udarbejdelse af Silicon nanotrådene Field-effekt Transistor for Kemiske og biosensorer Applications
Summary
We describe key steps for biosensing by using polysilicon nanowire field-effect transistors, including the preparation of the device and the immobilization and confirmation of a DNA molecular probe on the nanowire surface, as well as conditions for DNA sensing.
Abstract
Surveillance using biomarkers is critical for the early detection, rapid intervention, and reduction in the incidence of diseases. In this study, we describe the preparation of polycrystalline silicon nanowire field-effect transistors (pSNWFETs) that serve as biosensing devices for biomarker detection. A protocol for chemical and biomolecular sensing by using pSNWFETs is presented. The pSNWFET device was demonstrated to be a promising transducer for real-time, label-free, and ultra-high-sensitivity biosensing applications. The source/drain channel conductivity of a pSNWFET is sensitive to changes in the environment around its silicon nanowire (SNW) surface. Thus, by immobilizing probes on the SNW surface, the pSNWFET can be used to detect various biotargets ranging from small molecules (dopamine) to macromolecules (DNA and proteins). Immobilizing a bioprobe on the SNW surface, which is a multistep procedure, is vital for determining the specificity of the biosensor. It is essential that every step of the immobilization procedure is correctly performed. We verified surface modifications by directly observing the shift in the electric properties of the pSNWFET following each modification step. Additionally, X-ray photoelectron spectroscopy was used to examine the surface composition following each modification. Finally, we demonstrated DNA sensing on the pSNWFET. This protocol provides step-by-step procedures for verifying bioprobe immobilization and subsequent DNA biosensing application.
Introduction
Silicon Nanotråd felteffekttransistorer (SNWFETs) har fordelene ved ultra-høj følsomhed og direkte elektriske reaktioner på opladning variation af miljøet. I n-type SNWFETs for eksempel, når en negativt (eller positivt) ladet molekyle nærmer sig silicium nanotrådene (SNW), de luftfartsselskaber i SNW er udtømte (eller ophobes). Følgelig ledningsevnen af SNWFET aftager (eller stigninger) 1. Derfor kan detekteres nogen ladet molekyle nær SNW overflade af SNWFET enhed. Vitale biomolekyler herunder enzymer, proteiner, nukleotider og mange molekyler på celleoverfladen er ladningsbærere og kan overvåges ved hjælp SNWFETs. Med passende modifikationer, især immobilisering af en biomolekylært sonde på SNW, kan en SNWFET udvikles til en etiket-fri biosensor.
Overvågning ved hjælp biomarkører er afgørende for diagnosticering af sygdomme. Som det fremgår af tabel 1, har flere undersøgelser brugt NWFET-baserede biosensorer til etiket-fri, ultra-high-følsomhed og påvisning i realtid af forskellige biologiske mål, herunder et enkelt virus 2, adenosintriphosphat og kinase binding 3, neuronale signaler 4, metalioner 5,6, bakterielle toksiner 7, dopamin 8, DNA 9-11, RNA 12,13, enzym- og kræft biomarkører 14-19, menneskelige hormoner 20, og cytokiner 21,22. Disse undersøgelser har vist, at NWFET-baserede biosensorer et virksomt detektering platform for en bred vifte af biologiske og kemiske arter i en opløsning.
I SNWFET-baserede biosensorer, det immobiliserede på SNW enhedens overflade probe genkender en specifik biotarget. Immobilisere en bioprobe normalt indebærer en række trin, og det er afgørende, at hvert skridt er korrekt udført for at sikre et velfungerende biosensoren. Forskellige teknikker er blevet udviklet til at analysere sverflade sammensætning, herunder X-ray photoelectron spectroscopy (XPS), ellipsometri, trykvinkel måling, atomic force mikroskopi (AFM), og scanningselektronmikroskopi (SEM). Metoder såsom AFM og SEM direkte bevis på bioprobe immobilisering på nanotrådene enhed, mens metoder som XPS, ellipsometri, og kontakt vinkel måling er afhængige af parallelle forsøg udført på andre lignende materialer. I denne rapport beskriver vi bekræftelsen af hver ændring trin ved hjælp af to uafhængige metoder. XPS anvendes til at undersøge koncentrationen af bestemte atomer på polysilicon wafers, og variationer i de elektriske egenskaber af anordningen måles direkte at bekræfte ladningen variation på SNW overflade. Vi beskæftiger DNA biosensorer ved hjælp polykrystallinske SNWFETs (pSNWFETs) som et eksempel for at illustrere denne protokol. Immobilisering af en DNA-probe på SNW overflade består af tre trin: gruppe modifikation amin på det native hydroxyl overflade SNW, aldehyde gruppe modifikation, og 5'-aminomodified DNA-probe immobilisering. Ved hver ændring trin, kan enheden direkte detektere variationen i ladningen af den funktionelle gruppe immobiliseret på SNW overflade, fordi overfladeladninger forårsage lokale grænsefladespændinger potentielle ændringer over porten dielektriske som ændrer kanal strøm og konduktans 1. Afgifter omgiver SNW overflade kan elektrisk modulere de elektriske egenskaber af pSNWFET enhed; derfor overfladeegenskaberne af den SNW spiller en afgørende rolle i fastlæggelsen af de elektriske karakteristika af pSNWFET enheder. I de rapporterede procedurer kan immobilisering af en bioprobe på SNW overflade bestemmes direkte og bekræftet gennem elektrisk måling, og anordningen er forberedt til biosensorer applikationer.
Protocol
Representative Results
Discussion
Kommercialisering af top-down og bottom-up fremstilling tilgange til sSNWFETs anses vanskelig på grund af omkostningerne 32,33, SNW position kontrol 34,35, og dens lave produktion skala 36. Derimod fremstilling pSNWFETs er enkel og billig 37. Gennem top-down tilgang og kombination med sidevæggen spacer dannelse teknik (figur 1), kan størrelsen af SNW kontrolleres ved at justere varigheden af reaktiv plasmaætsning. Procedurerne for udarbejdelse af nanotråd…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
This research was financially supported by Ministry of Science and Technology, Taiwan (104-2514-S-009 -001, 104-2627-M-009-001 and 102-2311-B-009-004-MY3). We thank the National Nano Device Laboratories (NDL) for its valuable assistance during device fabrication and analysis.
Materials
| Acetone | ECHO | AH-3102 | |
| (3-Amonopropyl)triethoxysilane (APTES), ≥98% | Sigma-Aldrich | A3648 | Danger |
| Ethanol, anhydrous, 99.5% | ECHO | 484000203108A-72EC | |
| Glutaraldehyde solution (GA), 50% | Sigma-Aldrich | G7651 | Avoid light |
| Sodium cyanoborohydride, ≥95.0% | Fluka | 71435 | Danger and deliquescent |
| Sodium phosphate tribasic dodecahydrate, ≥98% | Sigma | 04277 | |
| Phosphoric acid, ≥99.0% | Fluka | 79622 | Deliquescent |
| Photoresist (iP3650) | Tokyo Ohka Kogyo Co., LTD | THMR-iP3650 HP | |
| Synthetic oligonucleotides, HPLC purified | Protech Technology | ||
| Tris(hydroxymethyl)aminomethane (Tris), ≥99.8% | USB | 75825 | |
| Keithley 2636 System SourceMeter | Keithley | ||
| SR830 DSP Lock-In Amplifier | Stanford Research Systems | ||
| SR570 Low-noise Current Preamplifier | Stanford Research Systems | ||
| Ni PXI Express | National Instruments |
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