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- W4382798889 abstract "This protocol outlines in detail how to measure illumination overhead (IO) and collect raw illumination intensity data using the Rigol ds1054z oscilloscope. In fluorescence microscopy, IO is defined as the time a sample is illuminated without the emitted light being captured by the camera, resulting in unnecessary photobleaching and phototoxicity. Minimizing IO is critical for all live imaging experiments. IO occurs due to delays/errors in the synchronization between the microscope fluorescence excitation light source, exposure of the sample to excitation light and the camera exposure time for capturing emitted fluorescence light. The amount of IO varies greatly across setups due to software settings, physical hardware and how components are connected and triggered by the hardware and/or software. Thus, it is recommended that each individual measure the IO of their own microscope and experimental setup including for their exact image acquisition settings. More details use of this protocol and the need to measure IO can be found in Kiepas et. al. JCS, 2020 (https://pubmed.ncbi.nlm.nih.gov/31988150/) and Kiepas et. al. Microscopy Today, 2020 (https://www.cambridge.org/core/journals/microscopy-today/article/microscope-hardware-and-software-delays-cause-phototoxicity/B964736E4617AEC17C8B1BBD8B4F37B8)" @default.
- W4382798889 created "2023-07-02" @default.
- W4382798889 creator A5068414716 @default.
- W4382798889 date "2023-06-08" @default.
- W4382798889 modified "2023-09-23" @default.
- W4382798889 title "How to Measure the Illumination Overhead (IO) of a Fluorescence Microscope v1" @default.
- W4382798889 doi "https://doi.org/10.17504/protocols.io.rm7vzxm35gx1/v1" @default.
- W4382798889 hasPublicationYear "2023" @default.
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