Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy.
Yicong WuAbhishek KumarCorey SmithEvan ArdielPanagiotis ChandrisRyan ChristensenIvan Rey-SuarezMin GuoHarshad D VishwasraoJiji ChenJianyong TangArpita UpadhyayaPatrick J La RiviereHari ShroffPublished in: Nature communications (2017)
Light-sheet fluorescence microscopy (LSFM) enables high-speed, high-resolution, and gentle imaging of live specimens over extended periods. Here we describe a technique that improves the spatiotemporal resolution and collection efficiency of LSFM without modifying the underlying microscope. By imaging samples on reflective coverslips, we enable simultaneous collection of four complementary views in 250 ms, doubling speed and improving information content relative to symmetric dual-view LSFM. We also report a modified deconvolution algorithm that removes associated epifluorescence contamination and fuses all views for resolution recovery. Furthermore, we enhance spatial resolution (to <300 nm in all three dimensions) by applying our method to single-view LSFM, permitting simultaneous acquisition of two high-resolution views otherwise difficult to obtain due to steric constraints at high numerical aperture. We demonstrate the broad applicability of our method in a variety of samples, studying mitochondrial, membrane, Golgi, and microtubule dynamics in cells and calcium activity in nematode embryos.
Keyphrases
- high resolution
- high speed
- single molecule
- mass spectrometry
- atomic force microscopy
- risk assessment
- induced apoptosis
- machine learning
- photodynamic therapy
- oxidative stress
- cell cycle arrest
- healthcare
- signaling pathway
- deep learning
- high throughput
- health risk
- health information
- cell death
- heavy metals
- fluorescence imaging
- cell proliferation
- pi k akt