Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses

被引:23
作者
Daaboul, George G. [1 ]
Freedman, David S. [1 ]
Scherr, Steven M. [2 ]
Carter, Erik [3 ]
Rosca, Alexandru [1 ]
Bernstein, David [1 ]
Mire, Chad E. [4 ,5 ,6 ]
Agans, Krystle N. [4 ,5 ,6 ]
Hoenen, Thomas [7 ,8 ]
Geisbert, Thomas W. [4 ,5 ,6 ]
Unlu, M. Selim [9 ,10 ,11 ]
Connor, John H. [3 ,10 ]
机构
[1] nanoView Diagnost Inc, Boston, MA USA
[2] Boston Univ, Dept Mech Engn, Boston, MA 02215 USA
[3] Boston Univ, Sch Med, Dept Microbiol, Boston, MA 02118 USA
[4] Galveston Natl Lab, Galveston, TX USA
[5] Dept Microbiol, Galveston, TX USA
[6] Univ Texas Med Branch, Immunol, Galveston, TX 77555 USA
[7] NIAID, LabVirol, Div Intramural Res, NIH,Rocky Mt Labs, Hamilton, MT 59840 USA
[8] Friedrich Loeffler Inst, Inst Mol Virol & Cell Biol, Greifswald, Germany
[9] Boston Univ, Dept Elect Engn, Boston, MA 02215 USA
[10] Boston Univ, Dept Biomed Engn, Boston, MA 02215 USA
[11] Boston Univ, Dept Phys, 590 Commonwealth Ave, Boston, MA 02215 USA
关键词
VESICULAR STOMATITIS-VIRUS; CRYOELECTRON TOMOGRAPHY; ELECTRON-MICROSCOPY; EBOLA-VIRUS; INFECTIVITY; VIRIONS; MASS;
D O I
10.1371/journal.pone.0179728
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Light microscopy is a powerful tool in the detection and analysis of parasites, fungi, and pro-karyotes, but has been challenging to use for the detection of individual virus particles. Unlabeled virus particles are too small to be visualized using standard visible light microscopy. Characterization of virus particles is typically performed using higher resolution approaches such as electron microscopy or atomic force microscopy. These approaches require purification of virions away from their normal millieu, requiring significant levels of expertise, and can only enumerate small numbers of particles per field of view. Here, we utilize a visible light imaging approach called Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) that allows automated counting and sizing of thousands of individual virions. Virions are captured directly from complex solutions onto a silicon chip and then detected using a reflectance interference imaging modality. We show that the use of different imaging wavelengths allows the visualization of a multitude of virus particles. Using Violet/UV illumination, the SP-IRIS technique is able to detect individual flavivirus particles (similar to 40 nm), while green light illumination is capable of identifying and discriminating between vesicular stomatitis virus and vaccinia virus (similar to 360 nm). Strikingly, the technology allows the clear identification of filamentous infectious ebolavirus particles and virus-like particles. The ability to differentiate and quantify unlabeled virus particles extends the usefulness of traditional light microscopy and can be embodied in a straightforward benchtop approach allowing widespread applications ranging from rapid detection in biological fluids to analysis of virus-like particles for vaccine development and production.
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页数:15
相关论文
共 29 条
[1]   Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography [J].
Bharat, Tanmay A. M. ;
Noda, Takeshi ;
Riches, James D. ;
Kraehling, Verena ;
Kolesnikova, Larissa ;
Becker, Stephan ;
Kawaoka, Yoshihiro ;
Briggs, John A. G. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2012, 109 (11) :4275-4280
[2]   Cryo-Electron Tomography of Marburg Virus Particles and Their Morphogenesis within Infected Cells [J].
Bharat, Tanmay A. M. ;
Riches, James D. ;
Kolesnikova, Larissa ;
Welsch, Sonja ;
Kraehling, Verena ;
Davey, Norman ;
Parsy, Marie-Laure ;
Becker, Stephan ;
Briggs, John A. G. .
PLOS BIOLOGY, 2011, 9 (11)
[3]   A new polymeric coating for protein microarrays [J].
Cretich, M ;
Pirri, G ;
Damin, F ;
Solinas, I ;
Chiari, M .
ANALYTICAL BIOCHEMISTRY, 2004, 332 (01) :67-74
[4]   Cryo-electron tomography of vaccinia virus [J].
Cyrklaff, M ;
Risco, C ;
Fernández, JJ ;
Jiménez, MV ;
Estéban, M ;
Baumeister, W ;
Carrascosa, JL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (08) :2772-2777
[5]   High-Throughput Detection and Sizing of Individual Low-Index Nanoparticles and Viruses for Pathogen Identification [J].
Daaboul, G. G. ;
Yurt, A. ;
Zhang, X. ;
Hwang, G. M. ;
Goldberg, B. B. ;
Uenlue, M. S. .
NANO LETTERS, 2010, 10 (11) :4727-4731
[6]   Digital Detection of Exosomes by Interferometric Imaging [J].
Daaboul, George G. ;
Gagni, Paola ;
Benussi, Luisa ;
Bettotti, Paolo ;
Ciani, Miriam ;
Cretich, Marina ;
Freedman, David S. ;
Ghidoni, Roberta ;
Ozkumur, Ayca Yalcin ;
Piotto, Chiara ;
Prosperi, Davide ;
Santini, Benedetta ;
Unlu, M. Selim ;
Chiari, Marcella .
SCIENTIFIC REPORTS, 2016, 6
[7]   Digital Sensing and Sizing of Vesicular Stomatitis Virus Pseudotypes in Complex Media: A Model for Ebola and Marburg Detection [J].
Daaboul, George G. ;
Lopez, Carlos A. ;
Chinnala, Jyothsna ;
Goldberg, Bennett B. ;
Connor, John H. ;
Unlue, M. Selim .
ACS NANO, 2014, 8 (06) :6047-6055
[8]  
Daaboul GG, 2014, ACS NANO
[9]   Filamentous influenza viruses [J].
Dadonaite, Bernadeta ;
Vijayakrishnan, Swetha ;
Fodor, Ervin ;
Bhella, David ;
Hutchinson, Edward C. .
JOURNAL OF GENERAL VIROLOGY, 2016, 97 :1755-1764
[10]   Properties of replication-competent vesicular stomatitis virus vectors expressing glycoproteins of filoviruses and arenaviruses [J].
Garbutt, M ;
Liebscher, R ;
Wahl-Jensen, V ;
Jones, S ;
Möller, P ;
Wagner, R ;
Volchkov, V ;
Klenk, HD ;
Feldmann, H ;
Ströher, U .
JOURNAL OF VIROLOGY, 2004, 78 (10) :5458-5465