Line-Shaped Illumination: A Promising Configuration for a Flexible Two-Photon Microscopy Setup
Source abstract
Abstract
An innovative two-photon microscope exploiting a line-shaped illumination has been recently devised and then implemented. Such configuration allows to carry out a real-time detection by means of standard CCD cameras and is able to maintain the same resolution as commonly used point-scanning devices, thus overcoming what is usually regarded as the main limitation of line-scanning microscopes. Here, we provide an overview of the applications in which this device has been tested and has proved to be a flexible and efficient tool, namely imaging of biological samples, in-depth sample reconstruction, two-photon spectra detection, and dye cross-section measurements. These results demonstrate that the considered setup is promising for future developments in many areas of research and applications.
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Research context
OpenAlex topics: Advanced Fluorescence Microscopy Techniques · Nonlinear Optical Materials Studies · Photoacoustic and Ultrasonic Imaging
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Dipartimento di Fisica & Astronomia, Università di Firenze, IT-50019 Sesto Fiorentino, FI, Italy
LENS—European Laboratory for Nonlinear Spectroscopy, IT-50019 Sesto Fiorentino, FI, Italy
INO-CNR—Istituto Nazionale di Ottica del Consiglio Nazionale delle Ricerche, IT-50019 Sesto Fiorentino, FI, Italy
Dipartimento di Ingegneria Industriale e dell’Informazione, Università di Pavia, IT-27100 Pavia, PV, Italy
Bright Solutions, IT-27100 Cura Carpignano, PV, Italy
References (23)
Show source-provided references
1. Two-Photon Excitation Fluorescence Microscopy10.1146/annurev.bioeng.2.1.399 ↗
2. Two-photon laser scanning fluorescence microscopy10.1126/science.2321027 ↗
3. Two-photon fluorescence correlation spectroscopy: Method and application to the intracellular environment10.1016/S0006-3495(95)80230-4 ↗
4. Analysis of efficiency of two-photon versus single-photon absorption for fluorescence generation in biological objects10.1046/j.1365-2818.1996.870647.x ↗
5. Advances in two photon scanning and scanless microscopy technologies for functional neural circuit imaging10.1109/JPROC.2016.2577380 ↗
6. Imaging techniques for harmonic and multiphoton absorption fluorescence microscopy10.1063/1.3184828 ↗
7. Two-photon absorption: An overview of measurements and principles10.1364/AOP.2.000451 ↗
8. Line scanning microscopy with two-photon fluorescence excitation. W-Pos. 192
9. Real-time two-photon confocal microsocopy using a femtosecond, amplified Ti:sapphire system10.1046/j.1365-2818.1996.97379.x ↗
10. Improved depth resolution in video-rate line-scanning multiphoton microscopy using temporal focusing10.1364/OL.30.001686 ↗
11. Fast calculation of the line-spread-function by transversal directions decoupling10.1088/2040-8978/18/7/075609 ↗
12. Tomaselli, A., Tartara, L., and Hasani, E. (2013). Two-Photon Excited Fluorescence Microscope. (WO/2014/154279), European Patent.
13. Measurement of two-photon-absorption spectra through nonlinear fluorescence produced by a line-shaped excitation beam10.1111/jmi.12675 ↗
14. Practical two-photon-absorption cross sections and spectra of Eosin and Hematoxylin10.1002/jbio.202000141 ↗
15. Hasani, E., Tartara, L., and Tomaselli, A. (2019, January 15–18). A novel technique for the measurement of twophoton absorption spectra of dyes for nonlinear fluorescence microscopy. Proceedings of the 2019 IEEE International Conference on BioPhotonics (BioPhotonics), Taipei, Taiwan.10.1109/ICB47650.2019.8976316 ↗
16. Hasani, E., Tartara, L., and Tomaselli, A. (2019, January 15–18). Line-scanning two-photon microscope for the investigation of biological samples. Proceedings of the 2019 IEEE International Conference on BioPhotonics (BioPhotonics), Taipei, Taiwan.10.1109/ICB47650.2019.8976315 ↗
17. Lipoprotein (a) Internalization by RAW 264.7 Cells is Associated with Morphological Changes and Accumulation of Lipids Detectable by Two-photon Scanning Microscopy10.9734/BJMMR/2016/27433 ↗
18. Advanced methods of microscope control using μManager software10.14440/jbm.2014.36 ↗
19. (2022, January 20). μManager Official Website. Available online: https://micro-manager.org/.
20. Sutherland, R.L. (2003). Handbook of Nonlinear Optics, Marcel Dekker. [2nd ed.].10.1201/9780203912539 ↗
21. (2020, April 10). Zipfel2020—Two-Photon Action Cross Sections of Alexa 488 Colorant, from Zipfel Lab Online Database of Cornell University. Available online: http://zipfellab.bme.cornell.edu/.
22. Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm10.1364/JOSAB.13.000481 ↗
23. Direct Measurement of Two-Photon Action Cross Section10.1021/acs.analchem.9b00392 ↗