TY - JOUR
T1 - Advanced easySTED microscopy based on two-photon excitation by electrical modulations of light pulse wavefronts
AU - Otomo, Kohei
AU - Hibi, Terumasa
AU - Fang, Yi Cheng
AU - Hung, Jui Hung
AU - Tsutsumi, Motosuke
AU - Kawakami, Ryosuke
AU - Yokoyama, Hiroyuki
AU - Nemoto, Tomomi
N1 - Funding Information:
MEXT/JSPS KAKENHI (JP16K15103, JP15H05953 “Resonance Bio”, and JP16H06280 “Advanced Bioimaging Support”); the Research Program of “Five-star Alliance” in “NJRC Mater. & Dev.” (MEXT); and Brain/MINDS, AMED, Japan.
Publisher Copyright:
© 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.
PY - 2018/6/1
Y1 - 2018/6/1
N2 - We developed a compact stimulated emission depletion (STED) two-photon excitation microscopy that utilized electrically controllable components. Transmissive liquid crystal devices inserted directly in front of the objective lens converted the STED light into an optical vortex while leaving the excitation light unaffected. Light pulses of two different colors, 1.06 and 0.64 μm, were generated by laser diode-based light sources, and the delay between the two pulses was flexibly controlled so as to maximize the fluorescence suppression ratio. In our experiments, the spatial resolution of this system was up to three times higher than that obtained without STED light irradiation, and we successfully visualize the fine microtubule network structures in fixed mammalian cells without causing significant photo-damage.
AB - We developed a compact stimulated emission depletion (STED) two-photon excitation microscopy that utilized electrically controllable components. Transmissive liquid crystal devices inserted directly in front of the objective lens converted the STED light into an optical vortex while leaving the excitation light unaffected. Light pulses of two different colors, 1.06 and 0.64 μm, were generated by laser diode-based light sources, and the delay between the two pulses was flexibly controlled so as to maximize the fluorescence suppression ratio. In our experiments, the spatial resolution of this system was up to three times higher than that obtained without STED light irradiation, and we successfully visualize the fine microtubule network structures in fixed mammalian cells without causing significant photo-damage.
UR - http://www.scopus.com/inward/record.url?scp=85048191801&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85048191801&partnerID=8YFLogxK
U2 - 10.1364/BOE.9.002671
DO - 10.1364/BOE.9.002671
M3 - Article
AN - SCOPUS:85048191801
VL - 9
SP - 2671
EP - 2680
JO - Biomedical Optics Express
JF - Biomedical Optics Express
SN - 2156-7085
IS - 6
M1 - #318596
ER -