TY - JOUR
T1 - Imaging subcellular dynamics with fast and lightefficient volumetrically parallelized microscopy
AU - Dean, Kevin M.
AU - Roudot, Philippe
AU - Welf, Erik S.
AU - Pohlkamp, Theresa
AU - Garrelts, Gerard
AU - Herz, Joachim
AU - Fiolka, Reto
N1 - Publisher Copyright:
© 2017 Optical Society of America.
PY - 2017/2/20
Y1 - 2017/2/20
N2 - In fluorescence microscopy, the serial acquisition of two-dimensional images to form a three-dimensional (3D) volume limits the maximum imaging speed. This is particularly evident when imaging adherent cells in a light-sheet fluorescence microscopy format, as their elongated morphologies require ~200 image planes per image volume. Here, by illuminating the specimen with three light sheets, each independently detected, we present a light-efficient, crosstalk- free, and volumetrically parallelized 3D microscopy technique that is optimized for high-speed (up to 14 Hz) subcellular (300 nm lateral, 600 nm axial resolution) imaging of adherent cells. We demonstrate 3D imaging of intracellular processes, including cytoskeletal dynamics in single-cell migration and collective wound healing for 1500 and 1000 time points, respectively. Furthermore, we capture rapid biological processes, including the trafficking of early endosomes with velocities exceeding 10 µm/s and calcium signaling in primary neurons.
AB - In fluorescence microscopy, the serial acquisition of two-dimensional images to form a three-dimensional (3D) volume limits the maximum imaging speed. This is particularly evident when imaging adherent cells in a light-sheet fluorescence microscopy format, as their elongated morphologies require ~200 image planes per image volume. Here, by illuminating the specimen with three light sheets, each independently detected, we present a light-efficient, crosstalk- free, and volumetrically parallelized 3D microscopy technique that is optimized for high-speed (up to 14 Hz) subcellular (300 nm lateral, 600 nm axial resolution) imaging of adherent cells. We demonstrate 3D imaging of intracellular processes, including cytoskeletal dynamics in single-cell migration and collective wound healing for 1500 and 1000 time points, respectively. Furthermore, we capture rapid biological processes, including the trafficking of early endosomes with velocities exceeding 10 µm/s and calcium signaling in primary neurons.
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U2 - 10.1364/OPTICA.4.000263
DO - 10.1364/OPTICA.4.000263
M3 - Article
C2 - 28944279
AN - SCOPUS:85013313968
SN - 2334-2536
VL - 4
SP - 263
EP - 271
JO - Optica
JF - Optica
IS - 2
ER -