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Ultra-localized single cell electroporation using silicon nanowires
KTH, Skolan för bioteknologi (BIO), Molekylär Bioteknologi.
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2013 (Engelska)Ingår i: Lab on a Chip, ISSN 1473-0197, E-ISSN 1473-0189, Vol. 13, nr 3, s. 336-339Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Analysis of cell-to-cell variation can further the understanding of intracellular processes and the role of individual cell function within a larger cell population. The ability to precisely lyse single cells can be used to release cellular components to resolve cellular heterogeneity that might be obscured when whole populations are examined. We report a method to position and lyse individual cells on silicon nanowire and nanoribbon biological field effect transistors. In this study, HT-29 cancer cells were positioned on top of transistors by manipulating magnetic beads using external magnetic fields. Ultra-rapid cell lysis was subsequently performed by applying 600-900 mV(pp) at 10 MHz for as little as 2 ms across the transistor channel and the bulk substrate. We show that the fringing electric field at the device surface disrupts the cell membrane, leading to lysis from irreversible electroporation. This methodology allows rapid and simple single cell lysis and analysis with potential applications in medical diagnostics, proteome analysis and developmental biology studies.

Ort, förlag, år, upplaga, sidor
2013. Vol. 13, nr 3, s. 336-339
Nyckelord [en]
Microfluidic Devices, Lysis, Manipulation, Potentials, Chip
Nationell ämneskategori
Biokemi och molekylärbiologi
Identifikatorer
URN: urn:nbn:se:kth:diva-117655DOI: 10.1039/c2lc40837fISI: 000312947300003Scopus ID: 2-s2.0-84872085096OAI: oai:DiVA.org:kth-117655DiVA, id: diva2:602809
Anmärkning

QC 20130204

Tillgänglig från: 2013-02-04 Skapad: 2013-02-01 Senast uppdaterad: 2017-12-06Bibliografiskt granskad
Ingår i avhandling
1. A Biotechnology Perspective on Silicon Nanowire FETs for Biosensor Applications
Öppna denna publikation i ny flik eller fönster >>A Biotechnology Perspective on Silicon Nanowire FETs for Biosensor Applications
2013 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

The study of silicon nanowire-FET-based electronic biosensor applications is an emerging scientific field. These biosensors have the benefit of being theoretically extremely sensitive and reports of down to femtomolar (fM) levels of biomolecule detection have been reported. This thesis is written from a biotechnological perspective on the development of a silicon nanowire-FET biosensor. The thesis project was oriented towards developing a novel affinity-based silicon nanowire-FET biosensor based on small (2-3 nm) protein affinity-binders denoted Affibody molecules. The hypothesis was that a smaller biological detector element would reduce the effect of Debye screening of the charged biomarker. This hypothesis was neither proved nor disproved, and a substantial amount of time and effort was spent on improving the function of the different biosensor components. In paper I, a study on the effect of the redox state and pH at solvent-to-surface interfaces of the reference gate electrodes was done by using solutions with alternating pH and varying ratios of the Fe(CN)63-/ Fe(CN)64- redox pair. These experiments showed that the selection of reference gate electrode has major implications on the signal readout in terms of false response and current instability. While a current drop due to potential change on the surface of a platinum reference electrode was observed, no such thing was observed using a silver/silver chloride reference gate electrode. The conclusion is that it is critical for performance to use a reference gate electrode that has a stable electrode potential such as silver/silver chloride. In paper II, a discovery was made when intending to use nanowire joule heating to lyse HT-29 and MCF-7 cells. Using fringing electric fields irreversible electroporation of a cell on top of a nanowire was achieved at 600-1200 mVpeak-to-peak at 10 MHz for 2 ms. The process was monitored using a 3,3´-dihexyloxacarbocyanine iodide (DiOC6(3)) and Propidium Iodide (PI) live-dead dye kit. The nanowire-mediated electroporation method releases the cell content without the risk of heat denaturation and it is ultra-localized. To address the concern on how to control and monitor organosilane monolayer formation in the surface functionalization of silicon nanowires, a microwave-assisted method was evaluated in paper III. Using ellipsometry, AFM, ATR-FTIR and fluorescence scanning it was shown that less than 10 minutes of incubation in 1% (v/v) APTES in toluene at 75⁰C is needed for formation of a 0.7 nm monolayer. In paper IV, surface functionalization was further explored by using microdispensing of solutions of capture probes for localized functionalization of individual devices for a multiplexed silicon nanowire-FET biosensor application. Besides showing by fluorescent scanning that oligonucleotide or protein spots of ~ 100 μm diameters could be deposited on individual silicon nanowires, the functionalization chemistry was validated by using the same protocol for immobilization of the Z domain from Staphylococcus aureus Protein A (SpA) on silicon dioxide-coated SPR sensor chips, followed by real-time detection of the binding of immunoglobulin G. The immunoglobulins as affinity-binders have a drawback due to large size and the importance of having the binding event near the device in silicon nanowire-FET biosensor due to the effect of Debye screening. In paper V, in an effort to further minimize the size of affinity-binders of potential value as capture probes in silicon nanowire-FET applications, a backbone-cyclized, minimized 2-helix affibody-molecule (ZHER2:342min) was designed and produced by Solid Phase Peptide Synthesis(SPPS). The 2-helix affibody-molecule was evaluated for in vivo molecular imaging of HER2-expressing tumours, which was demonstrated in mice carrying SKOV-3 xenografts.

Ort, förlag, år, upplaga, sidor
Stockholm: KTH Royal Institute of Technology, 2013. s. 84
Serie
Trita-BIO-Report, ISSN 1654-2312 ; 2013:18
Nationell ämneskategori
Nanoteknik
Identifikatorer
urn:nbn:se:kth:diva-133702 (URN)978-91-7501-889-8 (ISBN)
Disputation
2013-11-29, F1, Lindstedtsvägen 22, KTH, Stockholm, 10:00 (Engelska)
Opponent
Handledare
Anmärkning

QC 20131111

Tillgänglig från: 2013-11-11 Skapad: 2013-11-08 Senast uppdaterad: 2013-11-11Bibliografiskt granskad

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Eriksson Karlström, Amelie

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