scholarly journals How To Extend The Capabilities Of A Commercial Two-Photon Microscope To Perform Super-Resolution Imaging, Wavelength Mixing And Label-Free Microscopy.

2021 ◽  
Author(s):  
Chiara Peres ◽  
Chiara Nardin ◽  
Guang Yang ◽  
Fabio Mammano

Multimodal microscopy combines multiple non-linear techniques that take advantage of different optical processes to generate contrast and increase the amount of information that can be obtained from biological samples. However, the most advanced optical architectures are typically custom-made and require complex alignment procedures, as well as daily maintenance by properly trained personnel for optimal performance. Here, we describe a hybrid system we constructed to overcome these disadvantages by modifying a commercial upright microscope. We show that our multimodal imaging platform can be used to seamlessly perform two-photon STED, wavelength mixing and label-free microscopy in both ex vivo and in vivo samples. The system is highly stable and endowed with remote alignment hardware that ensures simplified operability for non-expert users. This optical architecture is an important step forward towards a wider practical applicability of non-linear optics to bioimaging.

2008 ◽  
Vol 92 (3) ◽  
pp. 359-365 ◽  
Author(s):  
R. Cicchi ◽  
L. Sacconi ◽  
A. Jasaitis ◽  
R.P. O’Connor ◽  
D. Massi ◽  
...  

2007 ◽  
Vol 15 (8) ◽  
pp. 4935 ◽  
Author(s):  
Sylvain Lecler ◽  
Stefan Haacke ◽  
Nhan Lecong ◽  
Olivier Crégut ◽  
Jean-Luc Rehspringer ◽  
...  

2011 ◽  
Vol 64 (9) ◽  
pp. 1269 ◽  
Author(s):  
Marek Samoc ◽  
T. Christopher Corkery ◽  
Andrew M. McDonagh ◽  
Marie P. Cifuentes ◽  
Mark G. Humphrey

The cubic hyperpolarizabilities of 1,3,5-(trans-[RuCl(dppe)2(C≡CC6H4-4-C≡C)])3C6H3 (1), 1,3,5-(trans-[Ru(C≡CPh)(dppe)2(C≡CC6H4-4-C≡C)])3C6H3 (2), 1,3,5-(trans-[Ru(C≡CC6H4-4-NO2)(dppe)2(C≡CC6H4-4-C≡C)])3C6H3 (3), 1,3,5-{trans-[Ru(C≡C-3,5-(trans-[Ru(C≡CPh)(dppe)2(C≡CC6H4-4-C≡C)])2C6H3)(dppe)2(C≡CC6H4-4-C≡C)]}3C6H3 (4), and 1,3,5-{trans-[Ru(C≡C-3,5-(trans-[Ru(C≡CC6H4-4-NO2)(dppe)2(C≡CC6H4-4-C≡C)])2C6H3)(dppe)2(C≡CC6H4-4-C≡C)]}3C6H3 (5) have been assessed over the spectral range 520–1600 nm using the Z-scan technique and ~150 fs pulses. All complexes exhibit negative values of γreal (corresponding to self-defocusing behaviour) and significant positive values of γimag (corresponding to two-photon absorption) at short wavelengths (up to 1000 nm). The maximal values of γreal and γimag increase in magnitude on dendrimer generation increase (proceeding from 2 to 4 or 3 to 5). The open-aperture Z-scan results have been used to confirm and contrast the two-photon (2PA) and three-photon absorption (3PA) behaviour of 1–5, the data being consistent with the existence of 2PA at the short wavelength range, but with significant 3PA at longer wavelengths for 1–3 and 5, a record 3PA coefficient for an inorganic complex for 5 at 1180 nm, and appreciable 3PA at the telecommunications wavelength of 1300 nm.


Symmetry ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 153 ◽  
Author(s):  
Christophe Humbert ◽  
Thomas Noblet

To take advantage of the singular properties of matter, as well as to characterize it, we need to interact with it. The role of optical spectroscopies is to enable us to demonstrate the existence of physical objects by observing their response to light excitation. The ability of spectroscopy to reveal the structure and properties of matter then relies on mathematical functions called optical (or dielectric) response functions. Technically, these are tensor Green’s functions, and not scalar functions. The complexity of this tensor formalism sometimes leads to confusion within some articles and books. Here, we do clarify this formalism by introducing the physical foundations of linear and non-linear spectroscopies as simple and rigorous as possible. We dwell on both the mathematical and experimental aspects, examining extinction, infrared, Raman and sum-frequency generation spectroscopies. In this review, we thus give a personal presentation with the aim of offering the reader a coherent vision of linear and non-linear optics, and to remove the ambiguities that we have encountered in reference books and articles.


Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 922
Author(s):  
William Querido ◽  
Shital Kandel ◽  
Nancy Pleshko

Advances in vibrational spectroscopy have propelled new insights into the molecular composition and structure of biological tissues. In this review, we discuss common modalities and techniques of vibrational spectroscopy, and present key examples to illustrate how they have been applied to enrich the assessment of connective tissues. In particular, we focus on applications of Fourier transform infrared (FTIR), near infrared (NIR) and Raman spectroscopy to assess cartilage and bone properties. We present strengths and limitations of each approach and discuss how the combination of spectrometers with microscopes (hyperspectral imaging) and fiber optic probes have greatly advanced their biomedical applications. We show how these modalities may be used to evaluate virtually any type of sample (ex vivo, in situ or in vivo) and how “spectral fingerprints” can be interpreted to quantify outcomes related to tissue composition and quality. We highlight the unparalleled advantage of vibrational spectroscopy as a label-free and often nondestructive approach to assess properties of the extracellular matrix (ECM) associated with normal, developing, aging, pathological and treated tissues. We believe this review will assist readers not only in better understanding applications of FTIR, NIR and Raman spectroscopy, but also in implementing these approaches for their own research projects.


2020 ◽  
Vol 223 (16) ◽  
pp. jeb214890
Author(s):  
Ebtesam Ali Barnawi ◽  
Justine E. Doherty ◽  
Patrícia Gomes Ferreira ◽  
Jonathan M. Wilson

ABSTRACTPotassium regulation is essential for the proper functioning of excitable tissues in vertebrates. The H+/K+-ATPase (HKA), which is composed of the HKα1 (gene: atp4a) and HKβ (gene: atp4b) subunits, has an established role in potassium and acid–base regulation in mammals and is well known for its role in gastric acidification. However, the role of HKA in extra-gastric organs such as the gill and kidney is less clear, especially in fishes. In the present study in Nile tilapia, Oreochromis niloticus, uptake of the K+ surrogate flux marker rubidium (Rb+) was demonstrated in vivo; however, this uptake was not inhibited with omeprazole, a potent inhibitor of the gastric HKA. This contrasts with gill and kidney ex vivo preparations, where tissue Rb+ uptake was significantly inhibited by omeprazole and SCH28080, another gastric HKA inhibitor. The cellular localization of this pump in both the gill and kidney was demonstrated using immunohistochemical techniques with custom-made antibodies specific for Atp4a and Atp4b. Antibodies against the two subunits showed the same apical ionocyte distribution pattern in the gill and collecting tubules/ducts in the kidney. Atp4a antibody specificity was confirmed by western blotting. RT-PCT was used to confirm the expression of both subunits in the gill and kidney. Taken together, these results indicate for the first time K+ (Rb+) uptake in O. niloticus and that HKA is implicated, as shown through the ex vivo uptake inhibition by omeprazole and SCH28080, verifying a role for HKA in K+ absorption in the gill's ionocytes and collecting tubule/duct segments of the kidney.


1995 ◽  
Vol 17 (11-12) ◽  
pp. 1555-1559 ◽  
Author(s):  
G. C. La Rocca ◽  
F. Bassani ◽  
V. M. Agranovich

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