Imaging tissue-mimic with light sheet microscopy: A comparative guideline.
Tissue mimics (TMs) on the scale of several hundred microns provide a beneficial cell culture configuration for in vitro engineered tissue and are currently under the spotlight in tissue engineering and regenerative medicine. Due to the cell density and size, TMs are fairly inaccessible to optical observation and imaging within these samples remains challenging. Light Sheet Fluorescence Microscopy (LSFM)- an emerging and attractive technique for 3D optical sectioning of large samples- appears to be a particularly well-suited approach to deal with them. In this work, we compared the effectiveness of different light sheet illumination modalities reported in the literature to improve resolution and/or light exposure for complex 3D samples. In order to provide an acute and fair comparative assessment, we also developed a systematic, computerized benchmarking method. The outcomes of our experiment provide meaningful information for valid comparisons and arises the main differences between the modalities when imaging different types of TMs.
ICFO-Institut de Ciences Fotonique, Av. Carl Friedrich Gauss, 3, 08860 Castelldefels, Barcelona, Spain.
Jordi
J
Andilla
ITAV, Université de Toulouse, CNRS, UPS, France.
Raphael
R
Jorand
ICFO-Institut de Ciences Fotonique, Av. Carl Friedrich Gauss, 3, 08860 Castelldefels, Barcelona, Spain.
Omar E
OE
Olarte
Institut Pasteur BioImage Analysis Unit, F-75015, Paris, France.|CNRS, UMR-3691, F-75015 Paris, France.
Alexandre C
AC
Dufour
ITAV, Université de Toulouse, CNRS, UPS, France.
Martine
M
Cazales
Institut Pasteur BioImage Analysis Unit, F-75015, Paris, France.|CNRS, UMR-3691, F-75015 Paris, France.
Yoann L E
YL
Montagner
Onera, The French Aerospace Lab, F-31100 Toulouse, France.
Romain
R
Ceolato
Onera, The French Aerospace Lab, F-31100 Toulouse, France.
Nicolas
N
Riviere
Institut Pasteur BioImage Analysis Unit, F-75015, Paris, France.|CNRS, UMR-3691, F-75015 Paris, France.
Jean-Christophe
JC
Olivo-Marin
ICFO-Institut de Ciences Fotonique, Av. Carl Friedrich Gauss, 3, 08860 Castelldefels, Barcelona, Spain.
Pablo
P
Loza-Alvarez
ITAV, Université de Toulouse, CNRS, UPS, France.
Corinne
C
Lorenzo
D000818:Animals; D032701:Biomimetics / Q000379:methods*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D032383:Myocytes, Cardiac / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods*; D051381:Rats; D059008:Time-Lapse Imaging