Single molecule fluorescence microscopy of carbon nanotubes

Single molecule microscopy has been extensively used in the past decade to study individual biomolecules with excellent spatial and temporal resolution. Meanwhile, characterization of nanomaterials and their development towards a variety of biological applications has progressed rapidly. Despite the comparable size of nanoparticles and biomolecules, no microscopy platform and technique currently exists to study the interactions of single biomolecules with nanoparticles. Here, we have successfully developed a set of experimental tools to study the interactions of DNA and proteins on a nanomaterial surface. We observe nucleic acid‐encapsulated carbon nanotubes by using fluorophore‐labeled complementary DNA to explore the sequence‐specific affinity of DNA to the nano‐surface. Our results demonstrate cooperative exfoliation of the oligonucleotides from the nanomaterial surface and sequence‐dependent bioavailability of nanotube‐adsorbed DNA for hybridization. The platform is employed in conjunction with a super resolution algorithm to pinpoint sites of DNA hybridization along the length of the nanotube. The ability of a microfluidic channel to easily exchange solution is used to study specific and non‐specific nuclease activity on nanotube‐adsorbed DNA. Protein function is mapped to local DNA topology on the nanomaterial and distance‐dependent arrest of protein activity is observed, resulting in a nanotube‐induced arrest of 60% protein activity within 1 nm from the nanoparticle. Accessibility of different points of contact between the DNA and nanotube are assayed for nuclease resistance and range from 5% to 50%.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Physics

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2012

Author dc:creator

• Jena, Prakrit

Contributors dc:contributor

• Ha, Taekjip
• Chemla, Yann R.
• Oono, Yoshitsugu
• Stack, John D.

• single molecule microscopy
• carbon nanotubes
• Fluorescence resonance energy transfer (FRET)
• Telomeric DNA
• DNA-SWNT
• Deoxyribonucleic acid (DNA)

Statement dc:rights

• Copyright 2012 Prakrit Vaibhav Jena

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/30972

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/30972