TY - JOUR
T1 - Light sheet-based fluorescence microscopy (LSFM) for the quantitative imaging of cells and tissues
AU - Pampaloni, Francesco
AU - Chang, Bo Jui
AU - Stelzer, Ernst H.K.
N1 - Funding Information:
The research of the authors was funded by the Cluster of Excellence for Macromolecular Complexes (CEF-MC, EXC-115) granted to the Goethe Universität Frankfurt am Main by the Deutsche Forschungsgemeinschaft (DFG) and by the German Ministry for Education and Research (BMBF, Forschungsschwerpunkt Biophotonik IV, project ProMEBS) and additionally by the National Science Council (NSC100-2917-I-564-030) in Taiwan, R.O.C. and EMBO (ASTF 404–2012).
Publisher Copyright:
© 2015, Springer-Verlag Berlin Heidelberg.
PY - 2015/4/1
Y1 - 2015/4/1
N2 - In light sheet-based fluorescence microscopy (LSFM), only the focal plane is illuminated by a laser light sheet. Hence, only the fluorophores within a thin volume of the specimen are excited. This reduces photo-bleaching and photo-toxic effects by several orders of magnitude compared with any other form of microscopy. Therefore, LSFM (aka single/selective-plane illumination microscopy [SPIM] or digitally scanned light sheet microscopy [DSLM]) is the technique of choice for the three-dimensional imaging of live or fixed and of small or large three-dimensional specimens. The parallel recording of millions of pixels with modern cameras provides an extremely fast acquisition speed. Recent developments address the penetration depth, the resolution and the recording speed of LSFM. The impact of LSFM on research areas such as three-dimensional cell cultures, neurosciences, plant biology and developmental biology is increasing at a rapid pace. The development of high-throughput LSFM is the next leap forward, allowing the application of LSFM in toxicology and drug discovery screening.
AB - In light sheet-based fluorescence microscopy (LSFM), only the focal plane is illuminated by a laser light sheet. Hence, only the fluorophores within a thin volume of the specimen are excited. This reduces photo-bleaching and photo-toxic effects by several orders of magnitude compared with any other form of microscopy. Therefore, LSFM (aka single/selective-plane illumination microscopy [SPIM] or digitally scanned light sheet microscopy [DSLM]) is the technique of choice for the three-dimensional imaging of live or fixed and of small or large three-dimensional specimens. The parallel recording of millions of pixels with modern cameras provides an extremely fast acquisition speed. Recent developments address the penetration depth, the resolution and the recording speed of LSFM. The impact of LSFM on research areas such as three-dimensional cell cultures, neurosciences, plant biology and developmental biology is increasing at a rapid pace. The development of high-throughput LSFM is the next leap forward, allowing the application of LSFM in toxicology and drug discovery screening.
KW - Cellular spheroids
KW - Digitally scanned light sheet-based microscopy
KW - High-throughput LSFM
KW - Light sheet-based fluorescence microscopy (LSFM)
KW - Single/selective-plane illumination microscopy
KW - Three-dimensional cell cultures
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U2 - 10.1007/s00441-015-2144-5
DO - 10.1007/s00441-015-2144-5
M3 - Review article
C2 - 25743693
AN - SCOPUS:84939989940
SN - 0302-766X
VL - 360
SP - 129
EP - 141
JO - Cell and Tissue Research
JF - Cell and Tissue Research
IS - 1
ER -