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BP: Fachverband Biologische Physik

BP 17: Bioimaging

BP 17.5: Invited Talk

Wednesday, March 20, 2024, 10:30–11:00, H 2032

Red photocontrollable fluorescent proteins in nanoscopy — •Francesca Pennacchietti — KTH Royal Institute of Technology, Stockholm, Sweden

The observation of organelles dynamics and macromolecular complex interactions inside living cells and tissues, requires minimally invasive imaging strategies. In this context, photocontrollable fluorescent proteins (FPs) play a crucial role as tags in optical super-resolution microscopy and functional live cell imaging. To this end we have previously shown that reversibly switchable FPs enable fast (1 Hz for a 50 x 50 um2) and gentler (< 1 kW/cm2) nanoscopy (Masullo et al, Nat Comm, 2018). Additionally, irreversibly photoconvertible FPs can achieve photolabeling with high spatiotemporal precision. Nevertheless, their photophysical complexity poses some challenges in expanding such techniques toward multiplexing and in vivo imaging. Here, we explore novel photoswitching mechanism for fluorescent proteins in the red and near-infrared region of the spectra and assess their compatibility with live cell imaging at the nanoscale (Pennacchietti et al, Nat. Meth, 2018). Finally, we present strategies to combine the spectral and photophysical fingerprint of distinct photocontrollable FPs to achieve multiplexing in live cell imaging at the nanoscale and photolabeling studies (Pennacchietti et al, Nat Comm, 2023).

Keywords: Photoswitching; live cell imaging; nanoscopy; multiplexing; near-infrared fluorescent proteins