Multi-photon fluorescence microscopy imaging of mitochondria in living cells excited by Yb-doped femtosecond fiber laser utilizing two- and three-photon competitive absorption

By employing an Yb-doped femtosecond fiber laser oscillating at 1030 nm, multi-photon fluorescence microscopy imaging was demonstrated, utilizing competitive two- and three-photon absorption processes for excitation of the fluorescence probes. Competitive two- and three-photon absorptions occurred f...

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Bibliographic Details
Published in:AIP advances 2020-09, Vol.10 (9), p.095219-095219-7
Main Authors: Moritomo, H., Onishi, S., Makino, Y., Matsumoto, K., Suzuki, Y., Kawamata, J.
Format: Article
Language:English
Subjects:
Competition
Cut off wavelength
Excitation
Fiber lasers
Fluorescent indicators
Image acquisition
Lasers
Microscopy
Mitochondria
Photon absorption
Photons
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Summary:By employing an Yb-doped femtosecond fiber laser oscillating at 1030 nm, multi-photon fluorescence microscopy imaging was demonstrated, utilizing competitive two- and three-photon absorption processes for excitation of the fluorescence probes. Competitive two- and three-photon absorptions occurred for all six of the fluorescence probes employed. The mitochondria of human embryonic kidney 293 cells were stained with the probes, and multi-photon fluorescence images were obtained in all cases. The brightest multi-photon fluorescence image was obtained when employing the probe compound possessing the longest cut-off wavelength. This can be attributed to the larger contribution of two-photon absorption to the competitive excitation process. Thus, by utilizing competitive two- and three-photon absorption processes, multi-photon fluorescence images can be acquired using Yb-doped femtosecond fiber laser excitation with a variety of probes.
ISSN:2158-3226
2158-3226
DOI:10.1063/5.0004953