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Imaging Technology Could Solve Childhood Disease Secrets

By MedImaging International staff writers
Posted on 14 Jan 2014
Image: A super-resolution optical image of a specific hRSV viral filament produced with dSTORM technology. The viral filament is approximately 4 microns in length, typical of hRSV (Photo courtesy of Eric Alonas and Philip Santangelo).
Image: A super-resolution optical image of a specific hRSV viral filament produced with dSTORM technology. The viral filament is approximately 4 microns in length, typical of hRSV (Photo courtesy of Eric Alonas and Philip Santangelo).
Image: Microscope image showing a cell infected with RSV. The RNA tagged by the probe is shown in red, while the nucleoprotein is green (Photo courtesy of Eric Alonas and Philip Santangelo).
Image: Microscope image showing a cell infected with RSV. The RNA tagged by the probe is shown in red, while the nucleoprotein is green (Photo courtesy of Eric Alonas and Philip Santangelo).
Most children, by the time they are two-years-old, have had respiratory syncytial virus (RSV) and suffered symptoms similar to a severe cold. However, for some children, in particular, for premature infants and those with underlying health problems, RSV can lead to pneumonia and bronchitis, which can require hospitalization and have long-term consequences.

A new technique for studying the structure of the RSV virion and the activity of RSV in living cells could help researchers solve the mysteries of the virus, including how it enters cells, how it replicates, how many genomes it inserts into its hosts, and possibly why specific lung cells escape the infection comparatively unharmed, which could provide scientists with the data 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 Dr. Philip Santangelo, an associate professor in the Wallace H. Coulter department of biomedical engineering at Georgia Institute of Technology (Georgia Tech; Atlanta, GA, USA) and Emory University (Atlanta, GA, USA). “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 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 Ebolavirus. “We’ve shown that we can tag the genome using our probes,” explained Dr. 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 nm, 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-µm 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 (Nashville, TN, USA) and Emory University, used a probe technology that quickly attaches to RNA within cells. The probe employs multiple fluorophores to indicate the presence of the viral RNA, allowing the researchers to see where it goes in host cells, and to view as infectious particles leave the cells to spread the infection. 

The research depended on a new method for labeling 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 tetravalently complexed to neutravidin. The chimeric combination was used to help the probes evade cellular defenses.

The probes, when introduced into cells, rapidly diffuse through a cell infected with RSV and bind to the virus’s RNA. Though binding tightly, the probe does not 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 green fluorescent protein (GFP) and gold nanoparticles.

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 tackle that movement, the laboratory of Thomas Barker, also from the Coulter department, used micropatterned fibronectin on glass to create 50-µm “islands” that contained the cells during the study.

Among the mysteries, that the researchers would like to solve 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,” Dr. 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.”

Related Links:

Georgia Institute of Technology
Emory University
Vanderbilt University
 

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80 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1
81 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1
82 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
83 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1
84 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
85 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
86 (34.809112548828): SELECT show_live FROM jos_com_show_setting LIMIT 1
87 (61.03515625): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750136 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
88 (40.054321289062): SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750136 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
89 (43.153762817383): SELECT id,name,title FROM jos_deeppockets WHERE cat=10 AND TYPE='file_folder' AND published=1 AND id != 294743300 AND id != 294743301 AND id != 294743302 AND id != 294743303 AND id != 294743304 AND id != 294743305 AND id != 294743306 AND id != 294743307 AND id != 294743308 AND id != 294743309 AND id != 294743310 AND metakey != 'aacc' ORDER BY `ordering`
90 (61.03515625): SELECT show_live FROM jos_com_show_setting LIMIT 1
91 (40.054321289062): SELECT show_live FROM jos_com_show_setting LIMIT 1
92 (36.954879760742): SELECT show_live FROM jos_com_show_setting LIMIT 1
93 (36.954879760742): SELECT show_live FROM jos_com_show_setting LIMIT 1
94 (47.206878662109): SELECT show_live FROM jos_com_show_setting LIMIT 1
95 (36.954879760742): SELECT show_live FROM jos_com_show_setting LIMIT 1
96 (41.961669921875): SELECT show_live FROM jos_com_show_setting LIMIT 1
97 (36.954879760742): SELECT show_live FROM jos_com_show_setting LIMIT 1
98 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
99 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1
100 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
101 (35.762786865234): SELECT show_live FROM jos_com_show_setting LIMIT 1
102 (36.001205444336): SELECT show_live FROM jos_com_show_setting LIMIT 1
103 (35.04753112793): SELECT show_live FROM jos_com_show_setting LIMIT 1
104 (88.930130004883): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
105 (62.942504882812): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
106 (91.075897216797): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750136 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
107 (68.187713623047): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750136 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
108 (104.18891906738): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
109 (77.962875366211): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
110 (68.902969360352): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.id=294750136 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
111 (666.85676574707): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.id=294750136 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
112 (738.14392089844): SELECT dp.name, dp.id, dp2.name as channel, ex.name as author_name, ex.specialization, ex.images FROM jos_deeppockets as dp LEFT JOIN jos_experts as ex ON ex.id = dp.author LEFT JOIN jos_deeppockets as dp2 ON dp2.id = dp.fid WHERE dp.author != 0 AND dp.cat = 10 AND dp.published = 1 AND ( (dp.fid = 294743300) OR (dp.fid = 294743301) OR (dp.fid = 294743302) OR (dp.fid = 294743303) OR (dp.fid = 294743304) OR (dp.fid = 294743305) OR (dp.fid = 294743306) OR (dp.fid = 294743307) OR (dp.fid = 294743308) OR (dp.fid = 294743309) OR (dp.fid = 294743310) ) ORDER BY RAND() LIMIT 1
113 (478.02925109863): SELECT dp.name, dp.id, dp2.name as channel, ex.name as author_name, ex.specialization, ex.images FROM jos_deeppockets as dp LEFT JOIN jos_experts as ex ON ex.id = dp.author LEFT JOIN jos_deeppockets as dp2 ON dp2.id = dp.fid WHERE dp.author != 0 AND dp.cat = 10 AND dp.published = 1 AND ( (dp.fid = 294743300) OR (dp.fid = 294743301) OR (dp.fid = 294743302) OR (dp.fid = 294743303) OR (dp.fid = 294743304) OR (dp.fid = 294743305) OR (dp.fid = 294743306) OR (dp.fid = 294743307) OR (dp.fid = 294743308) OR (dp.fid = 294743309) OR (dp.fid = 294743310) ) ORDER BY RAND() LIMIT 1
114 (152.1110534668): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
115 (65.088272094727): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
116 (75.101852416992): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
117 (52.928924560547): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
118 (61.988830566406): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
119 (54.836273193359): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
120 (57.93571472168): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
121 (54.121017456055): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
122 (351.90582275391): SELECT * FROM trending_articles WHERE `journal_id` = 10 AND `timestamp` <= NOW() + INTERVAL 7 DAY ORDER BY `order`
123 (!!!!! 30034.780502319): SELECT * FROM jos_deeppockets WHERE jos_deeppockets.cat = 10 AND jos_deeppockets.type = 'deeppockets_item' AND jos_deeppockets.published = 1 AND jos_deeppockets.created + INTERVAL 14 DAY >= NOW() ORDER BY jos_deeppockets.hits DESC LIMIT 7
124 (729.08401489258): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750136 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750136 AND type='deeppockets_item' AND published='1' LIMIT 1
125 (!!!!! 10412.931442261): ( SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.video != '' AND dp.video_source != '' AND dp.cat = 10 AND dp.published = 1 AND dp.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) UNION ( SELECT dpa.id, dpa.name, dpa.title, dpa.video, dpa.video_source, dpa.created FROM jos_deeppockets_archive AS dpa WHERE dpa.video != '' AND dpa.video_source != '' AND dpa.cat = 10 AND dpa.published = 1 AND dpa.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) ORDER BY created DESC LIMIT 3
126 (117.06352233887): ( SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.video != '' AND dp.video_source != '' AND dp.cat = 10 AND dp.published = 1 AND dp.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) UNION ( SELECT dpa.id, dpa.name, dpa.title, dpa.video, dpa.video_source, dpa.created FROM jos_deeppockets_archive AS dpa WHERE dpa.video != '' AND dpa.video_source != '' AND dpa.cat = 10 AND dpa.published = 1 AND dpa.type != 'deeppockets_item_link' ORDER BY created DESC LIMIT 3 ) ORDER BY created DESC LIMIT 3
127 (51.021575927734): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803337 LIMIT 1
128 (41.007995605469): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803337 LIMIT 1
129 (70.095062255859): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803276 LIMIT 1
130 (41.007995605469): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294803276 LIMIT 1
131 (391.00646972656): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets_archive AS dp WHERE dp.id = 294803276 LIMIT 1
132 (139.95170593262): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets_archive AS dp WHERE dp.id = 294803276 LIMIT 1
133 (42.915344238281): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294802958 LIMIT 1
134 (37.193298339844): SELECT dp.id, dp.name, dp.title, dp.video, dp.video_source, dp.created FROM jos_deeppockets AS dp WHERE dp.id = 294802958 LIMIT 1
135 (602.96058654785): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
136 (444.88906860352): SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
137 (251.05476379395): SELECT name FROM jos_deeppockets WHERE id=215
138 (46.014785766602): SELECT name FROM jos_deeppockets WHERE id=215
139 (758.88633728027): SELECT *, DATE_FORMAT(publish_up,'%d %b %Y') AS date_publish_up, DATE_FORMAT(publish_down,'%d %b %Y') AS date_publish_down FROM jos_events WHERE state=1 AND publish_up > NOW() and extra_info REGEXP '((;)|(^))215((;)|($))' AND catid IN (0,110) ORDER BY publish_up ASC LIMIT 3
140 (669.00253295898): SELECT *, DATE_FORMAT(publish_up,'%d %b %Y') AS date_publish_up, DATE_FORMAT(publish_down,'%d %b %Y') AS date_publish_down FROM jos_events WHERE state=1 AND publish_up > NOW() and extra_info REGEXP '((;)|(^))215((;)|($))' AND catid IN (0,110) ORDER BY publish_up ASC LIMIT 3
141 (92.029571533203): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
142 (61.988830566406): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
143 (!!!!! 8619.0700531006): TRUNCATE TABLE jos_trademed_module
144 (110.14938354492): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750136 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
145 (66.995620727539): SELECT dp.*, ch.title as channel, exp.name as author_name, exp.specialization as author_specialization, exp.images as author_photo FROM jos_deeppockets_archive as dp LEFT JOIN jos_experts as exp ON exp.id = dp.author LEFT JOIN jos_deeppockets as ch ON dp.fid = ch.id WHERE 1=1 AND dp.cat=10 AND dp.id=294750136 AND (dp.type='deeppockets_item' OR dp.type='deeppockets_item_link') AND dp.published='1' LIMIT 1
146 (828.98139953613): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
147 (53.882598876953): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND (type='deeppockets_item' OR type='deeppockets_item_link') AND published='1' LIMIT 1
148 (60.081481933594): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
149 (43.869018554688): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
150 (!!!!! 1185.8940124512): SELECT jos_item.* FROM jos_item INNER JOIN jos_number ON jos_item.numberID=jos_number.id WHERE 1=1 AND (jos_item.imgMini<>'') AND jos_item.type='L' AND jos_item.numberID= ( SELECT jos_number.id FROM jos_number WHERE jos_number.journalID IN (10) and jos_number.id in (select distinct jos_number.id from jos_number join jos_item on jos_item.numberid=jos_number.id where jos_item.type='P') ORDER BY (CONVERT(SUBSTRING(issue,1,2),SIGNED) + (CONVERT(SUBSTRING(issue,3,2),SIGNED)*100)) DESC LIMIT 0,1 ) ORDER BY RAND() DESC LIMIT 6
151 (66.041946411133): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
152 (49.114227294922): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
153 (54.121017456055): SELECT fid FROM jos_deeppockets WHERE cat=10 AND id=294750136 AND type='deeppockets_item' AND published='1' union SELECT fid FROM jos_deeppockets_archive WHERE cat=10 AND id=294750136 AND type='deeppockets_item' AND published='1' LIMIT 1
154 (54.121017456055): select distinct links.* from jos_textlinks links join jos_textlinks_placement placement on links.id=placement.link_id where journal_id=(10) and (channel_id=215 or channel_id is null)
155 (44.107437133789): select distinct links.* from jos_textlinks links join jos_textlinks_placement placement on links.id=placement.link_id where journal_id=(10) and (channel_id=215 or channel_id is null)
156 (51.975250244141): SELECT livesite FROM jos_dp_categories WHERE id=10 AND published='1' LIMIT 1
157 (197.17216491699): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
158 (77.962875366211): SELECT * FROM jos_deeppockets WHERE 1=1 AND id=294750136 AND published='1' union SELECT * FROM jos_deeppockets_archive WHERE 1=1 AND id=294750136 AND published='1' LIMIT 1
159 (36.954879760742): select jos_twitters.link from jos_twitters where cat=10
160 (33.855438232422): select jos_facebook.link from jos_facebook where cat=10
161 (30.994415283203): select jos_google_plus.link from jos_google_plus where cat=10
162 (28.848648071289): select jos_linkedin.link from jos_linkedin where cat=10
163 (331.16340637207): update jos_bannersmanager_banner set impmade=impmade+1 where id in(14202,14203)
164 (!!!!! 1157.9990386963): insert into jos_banners_delta values (14202,'2025-06-23 18:16:54','216.73.216.120','view'),(14203,'2025-06-23 18:16:54','216.73.216.120','view')