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Practical considerations for quantitative light sheet fluorescence microscopy
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DOI：10.1038/s41592-022-01632-x.png)
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This Perspective describes common pitfalls that can occur when using light sheet microscopy and offers guidance for improved quantitative imaging with these instruments. Fluorescence microscopy has evolved from a purely observational tool to a platform for quantitative, hypothesis-driven research. As such, the demand for faster and less phototoxic imaging modalities has spurred a rapid growth in light sheet fluorescence microscopy (LSFM). By restricting the excitation to a thin plane, LSFM reduces the overall light dose to a specimen while simultaneously improving image contrast. However, the defining characteristics of light sheet microscopes subsequently warrant unique considerations in their use for quantitative experiments. In this Perspective, we outline many of the pitfalls in LSFM that can compromise analysis and confound interpretation. Moreover, we offer guidance in addressing these caveats when possible. In doing so, we hope to provide a useful resource for life scientists seeking to adopt LSFM to quantitatively address complex biological hypotheses.
Keyword:
PLANE ILLUMINATION MICROSCOPY
ADAPTIVE OPTICS
BEGINNERS GUIDE
RESOLUTION
DECONVOLUTION
VIEW
REGISTRATION
EXCITATION
EMBRYOS
CELLS
期刊
IF:
32.1
论文数:
7.2K
被引数:
12.7W
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