Live spheroid formation recorded with light sheet-based fluorescence microscopy.
We provide a detailed protocol for a three-dimensional long-term live imaging of cellular spheroids with light sheet-based fluorescence microscopy. The protocol allows the recording of all phases of spheroid formation in three dimensions, including cell proliferation, aggregation, and compaction. We employ the human hepatic cell line HepaRG transfected with the fusion protein H2B-GFP, i.e., a fluorescing histone. The protocol allows monitoring the effect of drugs or toxicants.
Physical Biology Group, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe Universität Frankfurt am Main, Max-von-Laue-Str. 15, Frankfurt am Main, D-60438, Germany, francesco.pampaloni@physikalischebiologie.de.
Francesco
F
Pampaloni
Roli
R
Richa
Nariman
N
Ansari
Ernst H K
EH
Stelzer
D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics; D006657:Histones / Q000235:genetics; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D018874:Spheroids, Cellular / Q000648:ultrastructure*
D006657:Histones; D049452:Green Fluorescent Proteins