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

Boris Louis, Rafael Camacho, , Sergey Abakumov, Johannes Vandaele, Tetsuhiro Kudo, Hiroshi Masuhara, Ivan Scheblykin, Johan Hofkens & Susana Rocha
Multifocal plane microscopy allows capturing images at different focal planes simultaneously. Using a proprietary prism which splits the emitted light into paths of different length, 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...
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