Abstract
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 mu m(3). 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 mu m fluorescent beads into an optical trap. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
Original language | English |
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Article number | 401557 |
Pages (from-to) | 28656-28671 |
Number of pages | 16 |
Journal | Optics Express |
Volume | 28 |
Issue number | 19 |
DOIs | |
State | Published - 14 Sep 2020 |
Keywords
- 3-DIMENSIONAL PARTICLE TRACKING
- IMAGE CORRELATION SPECTROSCOPY
- MULTIFOCAL PLANE MICROSCOPY
- FLUORESCENCE MICROSCOPY
- LOCALIZATION MICROSCOPY
- MOLECULE LOCALIZATION
- CELL
- DYNAMICS
- DIFFUSION
- RECEPTOR