In Vivo Delivery of Nitric Oxide‐Sensing, Single‐Walled Carbon Nanotubes

Nicole M. Iverson1, Michael S. Strano2, Gerald N. Wogan1

1 Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, 2 Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts
Publication Name:  Current Protocols in Chemical Biology
Unit Number:   
DOI:  10.1002/9780470559277.ch140196
Online Posting Date:  June, 2015
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Detection of nitric oxide (NO) in vivo by single‐walled carbon nanotubes (SWNT) is based on the fluorescent properties of SWNT and the ability of NO to quench the fluorescence signal. Alterations of the signal can be utilized to detect a small molecule in vivo that has not previously been possible by other assay techniques. The protocols described here explain the techniques used to prepare NO‐detecting SWNTs and to administer them to mice by both intravenous and subcutaneous routes. These techniques can also be utilized with other SWNT sensors as well as non‐SWNT sensors. © 2015 by John Wiley & Sons, Inc.

Keywords: in vivo; single‐walled carbon nanotubes; nitric oxide; sensor; intravenous; subcutaneous; hydrogel

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Fabrication of Nitric Oxide Sensors Through DNA Wrapping of Single‐Walled Carbon Nanotubes
  • Basic Protocol 2: Hydrogel Encapsulation, Delivery and Detection of DNA‐Wrapped Single‐Walled Carbon Nanotubes
  • Basic Protocol 3: Pegylation, Intravenous Delivery, and Detection of DNA‐Wrapped Single‐Walled Carbon Nanotubes
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Fabrication of Nitric Oxide Sensors Through DNA Wrapping of Single‐Walled Carbon Nanotubes

  Materials
  • 5′ thiol‐modified d(AAAT) 7 (Integrated DNA Technologies)
  • SWNT (SouthWest NanoTechnologies, SG65i) tube diameter 0.77 ± 0.02 nm, aspect ratio >1,000, carbon content >95% by weight, (6,5) chirality >40%, and semiconducting content >95%
  • Sterile saline, 0.90% (w/v) NaCl in water
  • XPert Nano enclosure (Labconco)
  • Static gun (Sigma Aldrich)
  • Bath sonicator (Branson)
  • Ultrasonicator/tip sonicator (Qsonica, 1/8 inch microtip probe)
  • UV/Vis spectrophotometer

Basic Protocol 2: Hydrogel Encapsulation, Delivery and Detection of DNA‐Wrapped Single‐Walled Carbon Nanotubes

  Materials
  • PRONOVA SLM 20 alginate (250 mg; NovaMatrix)
  • Sterile SWNT solution (30 mg/liter, see protocol 1)
  • Sterile saline, 0.90% (w/v) NaCl in water
  • Slide‐A‐Lyzer MINI dialysis device, 2 K MWCO (Life Technologies)
  • Sterile BaCl 2 (0.1 M in water)
  • Depilatory agent (e.g., Nair bikini cream, sensitive formula)
  • Microcentrifuge tubes
  • Isoflurane and isoflurane delivery instrument
  • Razor
  • Sterile surgical equipment, including forceps, surgical scissors, 4‐0‐5‐0 polypropylene or nylon sutures, sterile drape, needle driver, betadine scrub, 70% isopropyl alcohol
  • Imaging instrument (e.g., Maestro, CRi™)

Basic Protocol 3: Pegylation, Intravenous Delivery, and Detection of DNA‐Wrapped Single‐Walled Carbon Nanotubes

  Materials
  • 5′ thiol‐modified d(AAAT) 7 (Integrated DNA Technologies)
  • Sterile saline, 0.90% (w/v) NaCl in water
  • SWNT (SouthWest NanoTechnologies, SG65i) tube diameter 0.77 ± 0.02 nm, aspect ratio >1,000, carbon content >95% by weight, (6,5) chirality >40%, and semiconducting content >95%
  • 0.5 M TCEP stock (see recipe)
  • Nanopure water
  • Methoxy‐PEG (5 kDa) maleimide (Sigma Aldrich)
  • PBS (phosphate buffered saline; Sigma Aldrich)
  • Depilatory agent (e.g., Nair bikini cream sensitive formula)
  • Isoflurane and isoflurane delivery instrument
  • Insulin syringes, 0.3 cc, 29 gauge, 0.5 inch (BD Biosciences)
  • Imaging instrument (e.g., Maestro, CRi™)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
  Heller, D.A., Jin, H., Martinez, B.M., Patel, D., Miller, B.M., Yeung, T.K., Jena, P.V., Hobartner, C., Ha, T., Silverman, S.K., and Strano, M.S. 2009. Multimodal optical sensing and analyte specificity using single‐walled carbon nanotubes. Nat. Nanotechnol. 4:114‐120.
  Iverson, N.M., Biskar, G., Farias, E., Ivanov, V., Wogan, G.N., and Strano, M.S. 2015. Quantitative tissue spectroscopy of near infrared fluorescent nanosensor implants. J. Biomed. Nanotechnol. In press.
  Iverson, N.M., Barone, P.W., Shandell, M., Trudel, L.J., Sen, S., Sen, F., Ivanov, V., Atolia, E., Farias, E., McNicholas, T.P., Reuel, N., Parry, N.M.A., Wogan, G.N., and Strano, M.S. 2013. In vivo biosensing via tissue localizable near infrared fluorescent single walled carbon nanotubes. Nat. Nanotechnol. 8:873‐880.
  Kim, J.H., Heller, D.A., Jin, H., Barone, P.W., Song, C., Zhang, J., Trudel, L.J., Wogan, G.N., Tannenbaum, S.R., and Strano, M.S. 2009. The rational design of nitric oxide selectivity in single‐walled carbon nanotube near‐infrared fluorescence sensors for biological detection. Nat. Chem. 1:473‐481.
  Wray, S., Cope, M., Delpy, D., Wyatt, J., and Reynolds, E. 1988. Characterization of the near infrared absorption spectra of cytochrome aa3 and haemoglobin for the non‐invasive monitoring of cerebral oxygenation. Biochim. Biophys. Acta 933:184‐192.
  Zhang, J.Q., Boghossian, A.A., Barone, P.W., Rwei, A., Kim, J.H., Lin, D.H., Heller, D.A., Hilmer, A.J., Nair, N., Reuel, N.F., and Strano, M.S. 2011. Single molecule detection of nitric oxide enabled by d(AT)(15) DNA adsorbed to near infrared fluorescent single‐walled carbon nanotubes. J. Am. Chem. Soc. 133:567‐581.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library