Light-sheet fluorescence imaging to localize cardiac lineage and protein distribution.
Light-sheet fluorescence microscopy (LSFM) serves to advance developmental research and regenerative medicine. Coupled with the paralleled advances in fluorescence-friendly tissue clearing technique, our cardiac LSFM enables dual-sided illumination to rapidly uncover the architecture of murine hearts over 10 by 10 by 10 mm3in volume; thereby allowing for localizing progenitor differentiation to the cardiomyocyte lineage and AAV9-mediated expression of exogenous transmembrane potassium channels with high contrast and resolution. Without the steps of stitching image columns, pivoting the light-sheet and sectioning the heart mechanically, we establish a holistic strategy for 3-dimentional reconstruction of the "digital murine heart" to assess aberrant cardiac structures as well as the spatial distribution of the cardiac lineages in neonates and ion-channels in adults.
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.|Department of Bioengineering, School of Engineering &Applied Sciences, UCLA, Los Angeles, CA 90095, USA.
Yichen
Y
Ding
Department of Bioengineering, School of Engineering &Applied Sciences, UCLA, Los Angeles, CA 90095, USA.
Juhyun
J
Lee
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.
Jianguo
J
Ma
Department of Bioengineering, School of Engineering &Applied Sciences, UCLA, Los Angeles, CA 90095, USA.
Kevin
K
Sung
Departments of Anesthesiology, Physiology and Medicine, Cardiovascular Research Laboratories, UCLA, Los Angeles, CA 90095, USA.
Tomohiro
T
Yokota
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.
Neha
N
Singh
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.
Mojdeh
M
Dooraghi
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.|Department of Bioengineering, School of Engineering &Applied Sciences, UCLA, Los Angeles, CA 90095, USA.
Parinaz
P
Abiri
Departments of Anesthesiology, Physiology and Medicine, Cardiovascular Research Laboratories, UCLA, Los Angeles, CA 90095, USA.
Yibin
Y
Wang
Department of Bioengineering, School of Engineering &Applied Sciences, UCLA, Los Angeles, CA 90095, USA.|Division of Dermatology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.|California NanoSystems Institute, UCLA, Los Angeles, CA 90095, USA.
Rajan P
RP
Kulkarni
Department of Molecular, Cellular and Developmental Biology, UCLA, Los Angeles, CA 90095, USA.|Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, UCLA, Los Angeles, CA 90095, USA.
Atsushi
A
Nakano
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.
Thao P
TP
Nguyen
School of Optical and Electronic Information, Huazhong University of Science and Technology, Wuhan, China.
Peng
P
Fei
Division of Cardiology, Department of Medicine, UCLA, Los Angeles, CA 90095, USA.|Department of Bioengineering, School of Engineering &Applied Sciences, UCLA, Los Angeles, CA 90095, USA.|California NanoSystems Institute, UCLA, Los Angeles, CA 90095, USA.
Tzung K
TK
Hsiai
D000818:Animals; D000831:Animals, Newborn; D002138:Calibration; D019070:Cell Lineage; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006352:Heart Ventricles / Q000166:cytology; D021621:Imaging, Three-Dimensional; D051379:Mice; D008856:Microscopy, Fluorescence; D009206:Myocardium / Q000166:cytology*; D015221:Potassium Channels / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*
D015221:Potassium Channels; D011506:Proteins; D049452:Green Fluorescent Proteins