Super-resolved FRET imaging by confocal fluorescence-lifetime single-molecule localization microscopy

FRET-based approaches are a unique tool for sensing the immediate surroundings and interactions of (bio)molecules. FRET imaging and FLIM (Fluorescence Lifetime Imaging Microscopy) enable the visualization of the spatial distribution of molecular interactions and functional states. However, conventional FLIM and FRET imaging provide average information over an ensemble of molecules within a diffraction-limited volume, which limits the spatial information, accuracy, and dynamic range of the observed signals. Here, we demonstrate an approach to obtain super-resolved FRET imaging based on single-molecule localization microscopy using an early prototype of a commercial time-resolved confocal microscope. DNA Points Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) with fluorogenic probes provides a suitable combination of background reduction and blinking kinetics compatible with the scanning speed of usual confocal microscopes. A single laser is used to excite the donor, a broad detection band is employed to retrieve both donor and acceptor emission, and FRET events are detected from lifetime information

DNA self-assembly of single molecules with deterministic position and orientation

International audienc

Becas de Verano 2019 del Instituto Balseiro.

El programa de Becas de Verano del Instituto Balseiro ofrece a estudiantes universitarios avanzados o recientemente egresados de carreras de grado en Ciencias o Ingenierías la posibilidad de realizar una pasantía durante el mes de febrero. El objetivo de la misma es familiarizarse con técnicas experimentales y colaborar en tareas de investigación en laboratorios del Centro Atómico Bariloche. Anualmente, 15 estudiantes son seleccionados y reciben ayuda económica completa. Cada participante elige un único tema de investigación entre una serie de propuestas y es guiado por un grupo de investigadores del Centro Atómico Bariloche. La pasantía finaliza con la entrega de un informe y la presentación de un póster en el que se detallan los resultados obtenidos a lo largo del mes de trabaj