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PLoS One. 2012;7(11):e49208. doi: 10.1371/journal.pone.0049208. Epub 2012 Nov 15.
Label-free single-particle imaging of the influenza virus by objective-type total internal reflection dark-field microscopy.
Enoki S, Iino R, Morone N, Kaihatsu K, Sakakihara S, Kato N, Noji H.
Source
Department of Applied Chemistry, School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo, Japan ; CREST, Japan Science and Technology Agency, Chiyoda-ku, Tokyo, Japan.
Abstract
Here we report label-free optical imaging of single particles of the influenza virus attached on a glass surface with a simple objective-type total internal reflection dark-field microscopy (TIRDFM). The capability of TIRDFM for the imaging of single viral particles was confirmed from fine correlation of the TIRDFM images with fluorescent immunostaining image and scanning electron microscopy image. The density of scattering spots in the TIRDFM images showed a good linearity against the virus concentration, giving the limit of detection as 1.2?10(4) plaque-forming units per milliliter. Our label-free optical imaging method does require neither elaborated sample preparation nor complex optical systems, offering a good platform for rapid and sensitive counting of viral particles.
PMID:
23166613
[PubMed - in process]
PMCID:
PMC3499535
Free full text
Label-free single-particle imaging of the influenza virus by objective-type total internal reflection dark-field microscopy.
Enoki S, Iino R, Morone N, Kaihatsu K, Sakakihara S, Kato N, Noji H.
Source
Department of Applied Chemistry, School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo, Japan ; CREST, Japan Science and Technology Agency, Chiyoda-ku, Tokyo, Japan.
Abstract
Here we report label-free optical imaging of single particles of the influenza virus attached on a glass surface with a simple objective-type total internal reflection dark-field microscopy (TIRDFM). The capability of TIRDFM for the imaging of single viral particles was confirmed from fine correlation of the TIRDFM images with fluorescent immunostaining image and scanning electron microscopy image. The density of scattering spots in the TIRDFM images showed a good linearity against the virus concentration, giving the limit of detection as 1.2?10(4) plaque-forming units per milliliter. Our label-free optical imaging method does require neither elaborated sample preparation nor complex optical systems, offering a good platform for rapid and sensitive counting of viral particles.
PMID:
23166613
[PubMed - in process]
PMCID:
PMC3499535
Free full text
