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Non-linear Spectral Microscopy-Multi-Photon Fl, SHG and THG
Published online by Cambridge University Press: 02 July 2020
Abstract
The non-linear nature of multi-photon fluorescence (FL) excitation, SHG and THG restricts the signal detecting volume to the vicinity of the focal point. As a result, the technology has intrinsic optical sectioning capability. The use of multi-photon fluorescence excitation also allows micro-fluorometry at high spatial resolution. Figure 1 shows a conventional optical micrograph of maize protoplasts, the time lapse fluorescence spectral change from a single chloroplast is shown in FIG 2. Under high intensity illumination, biological specimen not only emits fluorescence, but also generates harmonic emissions. in addition to the Ti-sapphire laser commonly used in multiphoton microscopy, the use of ultra-fast Cr-fosterite laser made simultaneous detecting two- and three-photon fluorescence, SHG and THG possible. in addition to the fluorescence signals generated by multi-photon excitation process, non-linear phenomena such as harmonic generation can also provide useful information about the structure and optical properties of a specimen (Kao et al., 2000). Simultaneous recording the spectral response in an image (x-y-λ) can provide insight about the nature of the signal.
- Type
- Challenges of Confocal Microscopy in the 21st Century (Organized by S. Paddock)
- Information
- Copyright
- Copyright © Microscopy Society of America 2001
Footnotes
Supported by National Science Council, Rep. of China: NSC88-2811-B-001-0023(PCC),NSC89-2811 -E-002-0058(PCC), NSC89-2311 -B-001 -032(BLL), NSC89-2112-M-l 10-016,NSC89-2216-E-110-003(FJK), NSC89-2215-E-002-064(CKS) and Academia Sinica (BLL).
References
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