Fast-tracking of single emitters in large volumes with nanometer precision

BORIS LOUIS, RAFAEL CAMACHO, ROGER BRESOLÍ-OBACH, SERGEY ABAKUMOV, JOHANNES VANDAELE, TESTSUHIRO KUDO, HIROSHI MASUHARA, IVAN G. SCHEBLYKIN, JOHAN HOFKENS, SUSANA ROCHA

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Resum

Multifocal plane microscopy allows for capturing images at different focal planes simultaneously. Using a proprietary prism which splits the emitted light into paths of different lengths, images at 8 different focal depths were obtained, covering a volume of 50x50x4 μm3. The position of single emitters was retrieved using a phasor-based approach across the different imaging planes, with better than 10 nm precision in the axial direction. We validated the accuracy of this approach by tracking fluorescent beads in 3D to calculate water viscosity. The fast acquisition rate (>100 fps) also enabled us to follow the capturing of 0.2 μm fluorescent beads into an optical trap.

Idioma originalAnglès
Número d’article401557
Pàgines (de-a)28656-28671
Nombre de pàgines16
RevistaOptics Express
Volum28
Número19
DOIs
Estat de la publicacióPublicada - de set. 2020

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