Abstract
The ability to perform optical sectioning is one of the great advantages of laser-scanning microscopy. This introduces, however, a number of difficulties due to the scanning process, such as lower frame rates due to the serial acquisition process. Here we show that by introducing spatiotemporal pulse shaping techniques to multiphoton microscopy it is possible to obtain full-frame depth resolved imaging completely without scanning. Our method relies on temporal focusing of the illumination pulse. The pulsed excitation field is compressed as it propagates through the sample, reaching its shortest duration at the focal plane, before stretching again beyond it. Combining temporal focusing with line-scanning microscopy results in an enhanced depth resolution, equivalent to that achieved by point scanning. Both the scanningless and the line-scanning techniques are applied to obtain depth-resolved two-photon excitation fluorescence (TPEF) images of drosophila egg-chambers.
| Original language | English |
|---|---|
| Article number | 10 |
| Pages (from-to) | 35-43 |
| Number of pages | 9 |
| Journal | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| Volume | 5700 |
| DOIs | |
| Publication status | Published - Jul 21 2005 |
| Event | Multiphoton Microscopy in the Biomedical Sciences V - San Jose, CA, United States Duration: Jan 23 2005 → Jan 25 2005 |
Keywords
- Laser-scanning
- Microscopy
- Nonlinear optics
- Two-photon excitation fluorescence
ASJC Scopus subject areas
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Atomic and Molecular Physics, and Optics
- Radiology Nuclear Medicine and imaging