Nanotechnology Now

Our NanoNews Digest Sponsors

Heifer International

Wikipedia Affiliate Button

Home > Press > Imaging technology could unlock mysteries of a childhood disease

This microscope image shows a cell infected with RSV. The RNA tagged by the probe is shown in red, while the nucleoprotein is green.

Credit: Image courtesy of Eric Alonas and Philip Santangelo
This microscope image shows a cell infected with RSV. The RNA tagged by the probe is shown in red, while the nucleoprotein is green.

Credit: Image courtesy of Eric Alonas and Philip Santangelo

Abstract:
By the time they're two, most children have had respiratory syncytial virus (RSV) and suffered symptoms no worse than a bad cold. But for some children, especially premature babies and those with underlying health conditions, RSV can lead to pneumonia and bronchitis - which can require hospitalization and have long-term consequences.

Imaging technology could unlock mysteries of a childhood disease

Atlanta, GA | Posted on December 30th, 2013

A new technique for studying the structure of the RSV virion and the activity of RSV in living cells could help researchers unlock the secrets of the virus, including how it enters cells, how it replicates, how many genomes it inserts into its hosts - and perhaps why certain lung cells escape the infection relatively unscathed. That could provide scientists information they need to develop new antiviral drugs and perhaps even a vaccine to prevent severe RSV infections.

"We want to develop tools that would allow us to get at how the virus really works," said Philip Santangelo, an associate professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University. "We really need to be able to follow the infection in a single living cell without affecting how the virus infects its hosts, and this technology should allow us to do that."

The research was supported by the National Institutes of Health's National Institute of General Medical Sciences and published online ahead of print in the journal ACS Nano on December 30, 2013. While RSV will be the first target for the work, the researchers believe the imaging technique they developed could be used to study other RNA viruses, including influenza and Ebola.

"We've shown that we can tag the genome using our probes," explained Santangelo. "What we've learned from this is that the genome does get incorporated into the virion, and that the virus particles created are infectious. We were able to characterize some aspects of the virus particle itself at super-resolution, down to 20 nanometers, using direct stochastic optical reconstruction microscopy (dSTORM) imaging."

RSV can be difficult to study. For one thing, the infectious particle can take different forms, ranging from 10-micron filaments to ordinary spheres. The virus can insert more than one genome into the host cells and the RNA orientation and structure are disordered, which makes it difficult to characterize.

The research team, which included scientists from Vanderbilt University and Emory University, used a probe technology that quickly attaches to RNA within cells. The probe uses multiple fluorophores to indicate the presence of the viral RNA, allowing the researchers to see where it goes in host cells - and to watch as infectious particles leave the cells to spread the infection.

"Being able to see the genome and the progeny RNA that comes from the genome with the probes we use really give us much more insight into the replication cycle," Santangelo said. "This gives us much more information about what the virus is really doing. If we can visualize the entry, assembly and replication of the virus, that would allow us to decide what to go after to fight the virus."

The research depended on a new method for labelling RNA viruses using multiply-labeled tetravalent RNA imaging probes (MTRIPS). The probes consist of a chimeric combination of DNA and RNA oligonucleotide labeled internally with fluorophores tetravelently complexed to neutravidin. The chimeric combination was used to help the probes evade cellular defenses.

"There are lots of sensors in the cell that look for foreign RNA and foreign DNA, but to the cell, this probe doesn't look like anything," Santangelo explained. "The cell doesn't see the nucleic acid as foreign."

Introduced into cells, the probes quickly diffuse through a cell infected with RSV and bind to the virus's RNA. Though binding tightly, the probe doesn't affect the normal activities of the virus and allows researchers to follow the activity for days using standard microscopy techniques. The MTRIPS can be used to complement other probe technology, such as GFP and gold nanoparticles.

Work done by graduate student Eric Alonas to concentrate the virus was essential to the project, Santangelo said. The concentration had to be done without adversely affecting the infectivity of the virus, which would have impacted its ability to enter host cells.

"It took quite a bit of work to get the right techniques to concentrate the RSV," he said. "Now we can make lots of infectious virus that's labelled and can be stored so we can use it when we want to."

To study the infection's progress in individual cells, the researchers faced another challenge: living cells move around, and following them complicates the research. To address that movement, the laboratory of Thomas Barker - also in the Coulter Department - used micro-patterned fibronectin on glass to create 50-micron "islands" that contained the cells during the study.

Among the mysteries that the researchers would like to tackle is why certain lung cells are severely infected - while others appear to escape ill effects.

"If you look at a field of cells, you see huge differences from cell to cell, and that is something that's not understood at all," Santangelo said. "If we can figure out why some cells are exploding with virus while others are not, perhaps we can figure out a way to help the bad ones look more like the good ones."

In addition to those already mentioned, the research team included James Crowe, professor of pediatrics at Vanderbilt University; Elizabeth Wright, assistant professor in the School of Medicine at Emory University; Daryll Vanover, Jeenah Jung, Chiara Zurla, Jonathan Kirschman, Vincent Fiore, and Alison Douglas from the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University; Aaron Lifland and Manasa Gudheti from Vutara Inc. in Salt Lake City, and Hong Yi from the Emory University School of Medicine.

One of the challenges of studying RSV is maintaining its activity in the laboratory setting - a problem parents of young children don't share.

"When you handle this virus in the lab, you have to always be careful about it losing infectivity," Santangelo noted. "But if you take a room full of children who have not been infected and let one infected child into the room, 15 minutes later all of the children will be infected."

###

The research described here was supported by the National Institute of General Medical Sciences of the National Institutes of Health under contract R01 GM094198-01. Any conclusions or opinions expressed are those of the authors and do not necessarily represent the official views of the NIH.

####

For more information, please click here

Contacts:
John Toon

404-894-6986

Copyright © Georgia Institute of Technology

If you have a comment, please Contact us.

Issuers of news releases, not 7th Wave, Inc. or Nanotechnology Now, are solely responsible for the accuracy of the content.

Bookmark:
Delicious Digg Newsvine Google Yahoo Reddit Magnoliacom Furl Facebook

Related Links

CITATION: Eric Alonas, et al., "Combining Single RNA Sensitive Probes with Subdiffraction-Limited and Live-Cell Imaging Enables the Characterization of Virus Dynamics in Cells," (ACS Nano, December 2013):

Related News Press

News and information

Leti to Demo Wristband with Embedded Sensors to Diagnose Sleep Apnea: APNEAband, Which Will Be Demonstrated at CES 2018, Also Monitors Mountain Sickness, Dehydration, Dialysis Treatment Response and Epileptic Seizures December 12th, 2017

Leti Develops World’s First Micro-Coolers for CERN Particle Detectors: Leti Design, Fabrication and Packaging Expertise Extends to Very Large Scientific Instruments December 11th, 2017

Untangling DNA: Researchers filter the entropy out of nanopore measurements December 8th, 2017

Device makes power conversion more efficient: New design could dramatically cut energy waste in electric vehicles, data centers, and the power grid December 8th, 2017

Imaging

JPK Instruments announce partnership with Swiss company, Cytosurge AG. The partnership makes Cytosurge’s FluidFM® technology available on the JPK NanoWizard® AFM platform December 8th, 2017

Researchers advance technique to detect ovarian cancer: Rice, MD Anderson use fluorescent carbon nanotube probes to achieve first in vivo success November 30th, 2017

Deben reports on a new publication from scientists at La Trobe University in Australia where their CT500 stage is used in micro scanning tomography experiments to better understand ceramic matrix composites under load November 29th, 2017

JPK reports on the exciting research in the School of Medicine at Sungkyunkwan University (SKKU), Suwon, South Korea using the NanoWizard® ULTRA Speed AFM to understand the binding of transcription factor Sox2 with super enhancers November 23rd, 2017

Govt.-Legislation/Regulation/Funding/Policy

Wheat gets boost from purified nanotubes: Rice University toxicity study shows plant growth enhanced by -- but only by -- purified nanotubes December 6th, 2017

Arrowhead Presents New Clinical Data Demonstrating a Sustained Host Response in Hepatitis B Patients Following RNAi Therapy — Up to 5.0 log10 reduction in HBsAg observed; data presented at HEP DART 2017 — December 6th, 2017

Chinese market opens up for Carbodeon nanodiamonds: Carbodeon granted Chinese Patent for Nanodiamond-containing Thermoplastic Thermal Compounds December 4th, 2017

Researchers advance technique to detect ovarian cancer: Rice, MD Anderson use fluorescent carbon nanotube probes to achieve first in vivo success November 30th, 2017

Nanomedicine

Leti to Demo Wristband with Embedded Sensors to Diagnose Sleep Apnea: APNEAband, Which Will Be Demonstrated at CES 2018, Also Monitors Mountain Sickness, Dehydration, Dialysis Treatment Response and Epileptic Seizures December 12th, 2017

Untangling DNA: Researchers filter the entropy out of nanopore measurements December 8th, 2017

Arrowhead Presents New Clinical Data Demonstrating a Sustained Host Response in Hepatitis B Patients Following RNAi Therapy — Up to 5.0 log10 reduction in HBsAg observed; data presented at HEP DART 2017 — December 6th, 2017

Copper will replace toxic palladium and expensive platinum in the synthesis of medications: The effectiveness of copper nanoparticles as a catalyst has been proven December 5th, 2017

Discoveries

UCLA chemists synthesize narrow ribbons of graphene using only light and heat: Tiny structures could be next-generation solution for smaller electronic devices December 8th, 2017

Untangling DNA: Researchers filter the entropy out of nanopore measurements December 8th, 2017

Device makes power conversion more efficient: New design could dramatically cut energy waste in electric vehicles, data centers, and the power grid December 8th, 2017

Wheat gets boost from purified nanotubes: Rice University toxicity study shows plant growth enhanced by -- but only by -- purified nanotubes December 6th, 2017

Announcements

Leti to Demo Wristband with Embedded Sensors to Diagnose Sleep Apnea: APNEAband, Which Will Be Demonstrated at CES 2018, Also Monitors Mountain Sickness, Dehydration, Dialysis Treatment Response and Epileptic Seizures December 12th, 2017

Leti Develops World’s First Micro-Coolers for CERN Particle Detectors: Leti Design, Fabrication and Packaging Expertise Extends to Very Large Scientific Instruments December 11th, 2017

Untangling DNA: Researchers filter the entropy out of nanopore measurements December 8th, 2017

Device makes power conversion more efficient: New design could dramatically cut energy waste in electric vehicles, data centers, and the power grid December 8th, 2017

Interviews/Book Reviews/Essays/Reports/Podcasts/Journals/White papers

UCLA chemists synthesize narrow ribbons of graphene using only light and heat: Tiny structures could be next-generation solution for smaller electronic devices December 8th, 2017

Device makes power conversion more efficient: New design could dramatically cut energy waste in electric vehicles, data centers, and the power grid December 8th, 2017

Creating a new kind of metallic glass December 7th, 2017

Copper will replace toxic palladium and expensive platinum in the synthesis of medications: The effectiveness of copper nanoparticles as a catalyst has been proven December 5th, 2017

Tools

Untangling DNA: Researchers filter the entropy out of nanopore measurements December 8th, 2017

JPK Instruments announce partnership with Swiss company, Cytosurge AG. The partnership makes Cytosurge’s FluidFM® technology available on the JPK NanoWizard® AFM platform December 8th, 2017

Researchers advance technique to detect ovarian cancer: Rice, MD Anderson use fluorescent carbon nanotube probes to achieve first in vivo success November 30th, 2017

Deben reports on a new publication from scientists at La Trobe University in Australia where their CT500 stage is used in micro scanning tomography experiments to better understand ceramic matrix composites under load November 29th, 2017

Research partnerships

Wheat gets boost from purified nanotubes: Rice University toxicity study shows plant growth enhanced by -- but only by -- purified nanotubes December 6th, 2017

Copper will replace toxic palladium and expensive platinum in the synthesis of medications: The effectiveness of copper nanoparticles as a catalyst has been proven December 5th, 2017

Researchers advance technique to detect ovarian cancer: Rice, MD Anderson use fluorescent carbon nanotube probes to achieve first in vivo success November 30th, 2017

Tiny robots step closer to treating hard-to-reach parts of the body November 25th, 2017

NanoNews-Digest
The latest news from around the world, FREE



  Premium Products
NanoNews-Custom
Only the news you want to read!
 Learn More
NanoTech-Transfer
University Technology Transfer & Patents
 Learn More
NanoStrategies
Full-service, expert consulting
 Learn More











ASP
Nanotechnology Now Featured Books




NNN

The Hunger Project