TY  - JOUR
TI  - Multicolor Stimulated Emission Depletion (STED) Microscopy to Generate High-resolution Images of Respiratory Syncytial Virus Particles and Infected Cells
AU  - Mehedi, Masfique  
AU  - Smelkinson, Margery 
AU  - Kabat, Juraj 
AU  - Ganesan, Sundar 
AU  - Collins, Peter L. 
AU  - Buchholz, Ursula J.
VL  - 7
IS  - 17
PY  - 2017
DA  - 2017/09/05
SP  - e2543
C1  - Bio-protocol 2017;7:e2543
DO  - 10.21769/BioProtoc.2543
UR  - https://doi.org/10.21769/BioProtoc.2543
AB  - Human respiratory syncytial virus (RSV) infection in human lung epithelial A549 cells induces filopodia, cellular protrusions consisting of F-actin, that extend to neighboring uninfected cells (Mehedi et al., 2016). High-resolution imaging via stimulated emission depletion (STED) microscopy revealed filamentous RSV particles along these filopodia, suggesting that filopodia facilitate RSV cell-to-cell spread (Mehedi et al., 2016). In this protocol, we describe how to fix, permeabilize, immunostain, and mount RSV-infected A549 cells for STED imaging. We show that STED increases resolution compared to confocal microscopy, which can be further improved by image processing using deconvolution software.
KW  - RSV
KW  - A549
KW  - STED microscopy
KW  - Filopodia
KW  - Cell-to-cell spread
KW  - Immunofluorescence
KW  - Confocal microscopy
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False