TY - JOUR
T1 - Achieving λ/10 resolution CW STED nanoscopy with a Ti:sapphire oscillator
AU - Liu, Yujia
AU - Ding, Yichen
AU - Alonas, Eric
AU - Zhao, Wenli
AU - Santangelo, Philip J.
AU - Jin, Dayong
AU - Piper, James A.
AU - Teng, Junlin
AU - Ren, Qiushi
AU - Xi, Peng
N1 - Copyright the Author(s) [2012]. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.
PY - 2012/6/27
Y1 - 2012/6/27
N2 - In this report, a Ti:Sapphire oscillator was utilized to realize synchronization-free stimulated emission depletion (STED) microscopy. With pump power of 4.6 W and sample irradiance of 310 mW, we achieved super-resolution as high as 71 nm. With synchronization-free STED, we imaged 200 nm nanospheres as well as all three cytoskeletal elements (microtubules, intermediate filaments, and actin filaments), clearly demonstrating the resolving power of synchronization-free STED over conventional diffraction limited imaging. It also allowed us to discover that, Dylight 650, exhibits improved performance over ATTO647N, a fluorophore frequently used in STED. Furthermore, we applied synchronization-free STED to image fluorescently-labeled intracellular viral RNA granules, which otherwise cannot be differentiated by confocal microscopy. Thanks to the widely available Ti:Sapphire oscillators in multiphoton imaging system, this work suggests easier access to setup super-resolution microscope via the synchronization-free STED.
AB - In this report, a Ti:Sapphire oscillator was utilized to realize synchronization-free stimulated emission depletion (STED) microscopy. With pump power of 4.6 W and sample irradiance of 310 mW, we achieved super-resolution as high as 71 nm. With synchronization-free STED, we imaged 200 nm nanospheres as well as all three cytoskeletal elements (microtubules, intermediate filaments, and actin filaments), clearly demonstrating the resolving power of synchronization-free STED over conventional diffraction limited imaging. It also allowed us to discover that, Dylight 650, exhibits improved performance over ATTO647N, a fluorophore frequently used in STED. Furthermore, we applied synchronization-free STED to image fluorescently-labeled intracellular viral RNA granules, which otherwise cannot be differentiated by confocal microscopy. Thanks to the widely available Ti:Sapphire oscillators in multiphoton imaging system, this work suggests easier access to setup super-resolution microscope via the synchronization-free STED.
UR - http://www.scopus.com/inward/record.url?scp=84862995094&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0040003
DO - 10.1371/journal.pone.0040003
M3 - Article
C2 - 22761944
AN - SCOPUS:84862995094
VL - 7
SP - 1
EP - 9
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 6
M1 - e40003
ER -