Nonlinear Structured Illumination Using a Fluorescent Protein Activating at the Readout Wavelength

Hui-Wen Lu-Walther, Wenya Hou, Martin Kielhorn, Yoshiyuki Arai, Takeharu Nagai, Michael M Kessels, Britta Qualmann and Rainer Heintzmann

PLOS ONE, 2016, vol. 11, issue 10, 1-14

Abstract: Structured illumination microscopy (SIM) is a wide-field technique in fluorescence microscopy that provides fast data acquisition and two-fold resolution improvement beyond the Abbe limit. We observed a further resolution improvement using the nonlinear emission response of a fluorescent protein. We demonstrated a two-beam nonlinear structured illumination microscope by introducing only a minor change into the system used for linear SIM (LSIM). To achieve the required nonlinear dependence in nonlinear SIM (NL-SIM) we exploited the photoswitching of the recently introduced fluorophore Kohinoor. It is particularly suitable due to its positive contrast photoswitching characteristics. Contrary to other reversibly photoswitchable fluorescent proteins which only have high photostability in living cells, Kohinoor additionally showed little degradation in fixed cells over many switching cycles.

Date: 2016
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Persistent link: https://EconPapers.repec.org/RePEc:plo:pone00:0165148

DOI: 10.1371/journal.pone.0165148

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