Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging
| Autor: | Aleksander Rebane, Caleb R. Stoltzfus |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Fluorescence-lifetime imaging microscopy Materials science Image quality 01 natural sciences Noise (electronics) Article law.invention 010309 optics 03 medical and health sciences Optics law 0103 physical sciences Image resolution ocis:(170.0170) Medical optics and biotechnology ocis:(140.7090) Ultrafast lasers ocis:(000.4430) Numerical approximation and analysis business.industry ocis:(230.4320) Nonlinear optical devices ocis:(120.0120) Instrumentation measurement and metrology Laser ocis:(190.4360) Nonlinear optics devices Atomic and Molecular Physics and Optics 030104 developmental biology Excited state Femtosecond Optoelectronics business Ultrashort pulse Biotechnology |
| Zdroj: | Biomedical Optics Express |
| ISSN: | 2156-7085 |
| Popis: | We study the optimal conditions for high throughput two-photon excited fluorescence (2PEF) and three-photon excited fluorescence (3PEF) imaging using femtosecond lasers. We derive relations that allow maximization of the rate of imaging depending on the average power, pulse repetition rate, and noise characteristics of the laser, as well as on the size and structure of the sample. We perform our analysis using ~100 MHz, ~1 MHz and 1 kHz pulse rates and using both a tightly-focused illumination beam with diffraction-limited image resolution, as well loosely focused illumination with a relatively low image resolution, where the latter utilizes separate illumination and fluorescence detection beam paths. Our theoretical estimates agree with the experiments, which makes our approach especially useful for optimizing high throughput imaging of large samples with a field-of-view up to 10x10 cm(2). |
| Databáze: | OpenAIRE |
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