Two-Color, Two-Photon Imaging at Long Excitation Wavelengths Using a Diamond Raman Laser

Trägårdh, Johanna; Murtagh, Michelle; Robb, G. R. M.; Parsons, Maddy; Lin, Jipeng; Spence, David J.; McConnell, Gail

doi:10.1017/s143192761601151x

Cited by 6 publications

(4 citation statements)

References 24 publications

(34 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To date, diamond Raman lasers have seen little use in microscopy. A Ti:S pumped diamond Raman laser has been used for asynchronous multi-color imaging in thin cell samples 24 , and a Nd:YVO 4 -pumped, picosecond diamond Raman laser emitting at 1240 nm has been demonstrated 25 , but not used for imaging. We show, for the first time, that a diamond Raman laser can generate femtosecond (~100 fs) pulse-widths at 1240 nm through pumping with an amplified ytterbium fiber source, and that the output is stable enough for routine use as a microscopy source.…”

Section: Introductionmentioning

confidence: 99%

Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

et al. 2017

View full text Add to dashboard Cite

Two-color multiphoton microscopy through wavelength mixing of synchronized lasers has been shown to increase the spectral window of excitable fluorophores without the need for wavelength tuning. However, most currently available dual output laser sources rely on the costly and complicated optical parametric generation approach. In this report, we detail a relatively simple and low cost diamond Raman laser pumped by a ytterbium fiber amplifier emitting at 1055 nm, which generates a first Stokes emission centered at 1240 nm with a pulse width of 100 fs. The two excitation wavelengths of 1055 and 1240 nm, along with the effective two-color excitation wavelength of 1140 nm, provide an almost complete coverage of fluorophores excitable within the range of 1000–1300 nm. When compared with 1055 nm excitation, two-color excitation at 1140 nm offers a 90% increase in signal for many far-red emitting fluorescent proteins (for example, tdKatushka2). We demonstrate multicolor imaging of tdKa-tushka2 and Hoechst 33342 via simultaneous two-color two-photon, and two-color three-photon microscopy in engineered 3D multicellular spheroids. We further discuss potential benefits and applications for two-color three-photon excitation. In addition, we show that this laser system is capable of in vivo imaging in mouse cortex to nearly 1 mm in depth with two-color excitation.

show abstract

Section: Introductionmentioning

confidence: 99%

Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

et al. 2017

View full text Add to dashboard Cite

show abstract

“…As demonstrated in Section 2, the 2PE wavelength at 1047 nm overlaps with the absorption band of several bio-imaging dyes. However, both Cr:Forsterite and Nd:YLF mode-locked lasers lack a wide wavelength tuning range (Trägårdh et al, 2016), which implies that fluorescent probes must be carefully chosen in order to match the small bandwidths, therefore this tends to limit biological imaging to just one colour. An example of a solid-state laser being used for in vivo applications involve a Nd:YLF laser being used to perform 2P NIR-II imaging of a stained zebrafish embryo (Wokosin et al, 1996a) (Section 3.2, Table 3).…”

Section: Nir-ii Solid-state Pulsed Lasersmentioning

confidence: 99%

“…To overcome the drawback of limited excitation wavelengths of such solid-state lasers, multi-colour 2P imaging can instead be achieved by using two tunable lasers in tandem, although this is a relatively high-cost solution. Other solutions have been found in utilising the second Stokes shift to extend output wavelengths in Ti:Sapphire laser systems, or in methods such as phase-shaping (Brenner et al, 2013;Trägårdh et al, 2016).…”

Section: Nir-ii Solid-state Pulsed Lasersmentioning

confidence: 99%

Two-Photon Absorption: An Open Door to the NIR-II Biological Window?

et al. 2022

View full text Add to dashboard Cite

The way in which photons travel through biological tissues and subsequently become scattered or absorbed is a key limitation for traditional optical medical imaging techniques using visible light. In contrast, near-infrared wavelengths, in particular those above 1000 nm, penetrate deeper in tissues and undergo less scattering and cause less photo-damage, which describes the so-called “second biological transparency window”. Unfortunately, current dyes and imaging probes have severely limited absorption profiles at such long wavelengths, and molecular engineering of novel NIR-II dyes can be a tedious and unpredictable process, which limits access to this optical window and impedes further developments. Two-photon (2P) absorption not only provides convenient access to this window by doubling the absorption wavelength of dyes, but also increases the possible resolution. This review aims to provide an update on the available 2P instrumentation and 2P luminescent materials available for optical imaging in the NIR-II window.

show abstract

“…Currently, multi-color two-photon microscopy imaging is a research hotspot in biomedical photonics, the reported multi-color two-photon microscopy mainly includes the following implementation methods. On one hand, multiple femtosecond lasers are used as excitation sources to excite different fluorescence probes, however, it will make the optical path of the imaging system complicated, expensive, and reduce the stability of the system [7][8][9]. On the other hand, a new type of pulsed laser can be developed as a light source, which can simultaneously excite a variety of fluorescent dyes.…”

Section: Introductionmentioning

confidence: 99%

Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation

et al. 2022

View full text Add to dashboard Cite

Two-photon probes with broad absorption spectra are beneficial for multi-color two-photon microscopy imaging, which is one of the most powerful tools to study the dynamic processes of living cells. To achieve multi-color two-photon imaging, multiple lasers and detectors are usually required for excitation and signal collection, respectively. However, one makes the imaging system more complicated and costly. Here, we demonstrate a multi-color two-photon imaging method with a single-wavelength excitation by using a signal separation strategy. The method can effectively solve the problem of spectral crosstalk by selecting a suitable filter combination and applying image subtraction. The experimental results show that the two-color and three-color two-photon imaging are achieved with a single femtosecond laser. Furthermore, this method can also be combined with multi-photon imaging technology to reveal more information and interaction in thick biological tissues.

show abstract

Two-Color, Two-Photon Imaging at Long Excitation Wavelengths Using a Diamond Raman Laser

Abstract: Microscopy and Microanalysis, a Cambridge University Press journalthe Ti:Sapphire laser, resulting in a laser system tunable 680 nm-1200 nm it can be used for twophoton excitation of a large variety and combination of dyes.

Cited by 6 publications

References 24 publications

Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

Two-Photon Absorption: An Open Door to the NIR-II Biological Window?

Multi-Color Two-Photon Microscopic Imaging Based on a Single-Wavelength Excitation

Contact Info

Product

Resources

About