Two-photon microscopy (2PM) performs a reliably environment friendly position in noninvasive deep-tissue imaging in biomedical investigation. Since the invention of the two-photon microscope on the finish of the twentieth century, there was a gentle circulate of associated analysis advancing 2PM—from fluorophores to imaging strategies and purposes—within the fields of biochemistry and drugs.
How to picture deeper tissues has been an important problem in two-photon imaging. Laser sources can probably handle that problem. However, the standard Ti:sapphire mode-locked laser for two-photon imaging is proscribed by its excessive repetition frequency and can’t present the excessive pulse vitality wanted for deep tissue imaging at low publicity energy. The fiber laser overcomes the excessive repetition price conveniently by including a couple of dozen meters of fibers within the cavity, however in some circumstances it suffers from low positive aspects and low signal-to-noise ratio (SNR).
Recently, as reported in Advanced Photonics Nexus, researchers from Kenneth Wong’s Omega Group at University of Hong Kong (HKU) developed a high-performance laser as a brand new sort of gentle supply for multiphoton microscopy. They reported a 937-nm laser, frequency doubled from an all-fiber mode-locked laser at 1.8 μm, with a low repetition price of ~9 MHz and a excessive SNR of 74 dB.
The novel 937-nm laser supply depends on self-phase modulation within the single-mode fiber to concurrently enhance the 1.8 μm energy and compress the heartbeat width. The 937-nm laser design is appropriate for high-sensitivity deep-tissue imaging of a number of fluorescence proteins. The laser gentle supply gives two-photon excitations on a number of organic tissue varieties. The penetration depth demonstrated with a mouse mind reached 620 μm, revealing the aptitude of this method for deep tissue imaging. The researchers additionally performed second-harmonic era (SHG) imaging, demonstrating label-free imaging and initially validating the potential of this gentle supply for multimodal imaging purposes.
Thanks to its low repetition frequency and excessive signal-to-noise ratio, the sunshine supply requires solely 10 mW of energy to picture tissue at depths of over 600 µm, considerably decrease than the 40-MHz fiber laser, which requires roughly 200-mW energy at an analogous depth. This enormously reduces photobleaching and photodamage in imaging, enhancing the depth of tissue imaging and security in dwell (in vivo) imaging.
This work will facilitate higher insights into deep tissue imaging for analysis and biomedical purposes. Postdoctoral fellow at HKU and corresponding creator Tian Qiao remarks that “this novel high-SNR 937-nm laser supply achieves an excellent steadiness between sensitivity, penetration depth, and imaging speed for two-photon imaging. Its great performance in two-photon imaging indicates its exciting potential for biological investigations, such as in vivo deep-tissue imaging and multimode imaging.”
Hongsen He et al, Deep-tissue two-photon microscopy with a frequency-doubled all-fiber mode-locked laser at 937 nm, Advanced Photonics Nexus (2022). DOI: 10.1117/1.APN.1.2.026001
High-performance 937-nm laser lets scientists see deeper with decrease energy (2022, September 13)
retrieved 13 September 2022
This doc is topic to copyright. Apart from any truthful dealing for the aim of personal examine or analysis, no
half could also be reproduced with out the written permission. The content material is supplied for info functions solely.