The role of tissue fluorescence in in vivo optical bioimaging

The following article appeared in Journal of Applied Physics 128.17 (2020): 171101 and may be found at https://doi.org/10.1063/5.0021854The technological advancements made in optics and semiconductors (e.g., cameras and laser diodes) working with infrared have brought interest in optical bioimaging back to the forefront of research investigating in vivo medical imaging techniques. The definition of the near-infrared transparency windows has turned optical imaging into more than just a method for topical imaging applications. Moreover, this has focused attention back to tissue fluorescence, emissions by tissues and organs that occur when excited by external illumination sources. Most endogenous fluorophores emit in the blue to green range of the electromagnetic spectrum and the resulting tissue fluorescence can be employed in studies from cells to tissue metabolism or avoided by shifting to the red if seen as unwanted autofluorescence. With the more recent move to infrared, it was discovered that autofluorescence is not limited to the visible but also strongly affects in vivo imaging in the infrared. In this Tutorial, we give an overview on tissue fluorescence and tissue interactions with excitation light as well as their effect on in vivo imaging. Furthermore, potential sources of tissue fluorescence in the near-infrared are identified and we describe approaches for successful biomedical imaging in the biological windows, taking into consideration infrared autofluorescence and summarizing techniques for avoiding it in in vivo imaging experimentsThis work was supported by the Spanish Ministry of Economy and Competitiveness under Project No. MAT2016-75362-C3-1-R, the Spanish Ministry of Sciences, Innovation and Universities under Project No. PID2019-106211RB-I00 (NANONERV), by the Instituto de Salud Carlos III (Nos. PI16/00812 and PI19/00565), and through the Comunidad Autónoma de Madrid (No. B2017/ BMD-3867RENIMCM), and co-financed by the European Structural and investment fund. Additional funding was provided by the European Union’s Horizon 2020 FET Open project NanoTBTech (Grant Agreement No. 801305), the Fundación para la Investigación Biomédica del Hospital Universitario Ramón y Cajal under Project No. IMP18_38(2018/0265), and also COST action CA17140. Y.S. acknowledges a scholarship from the China Scholarship Council (No.201806870023), E.X. is grateful for a Juan de la Cierva Formación scholarship (No. FJC2018-036734-I), and D.H.O. is thankful to the Instituto de Salud Carlos III for a Sara Borrell Fellowship (No. CD17/00210). The authors thank Dr. Blanca del Rosal for the helpful discussion and input on the manuscrip

The near-infrared autofluorescence fingerprint of the brain

This is the peer reviewed version of the following article: Lifante, J, del Rosal, B, Chaves-Coira, I, Fernández, N, Jaque, D, Ximendes, E. The near-infrared autofluorescence fingerprint of the brain. J. Biophotonics. 2020; 13:e202000154, which has been published in final form at https://doi.org/10.1002/jbio.202000154. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived VersionsThe brain is a vital organ involved in mostof the central nervous system disorders.Their diagnosis and treatment require fast,cost-effective, high-resolution and high-sensitivity imaging. The combinationof a new generation of luminescent nanoparticles and imaging systems work-ing in the second biological window (near-infrared II [NIR-II]) is emerging asa reliable alternative. For NIR-II imaging to become a robust technique at thepreclinical level, full knowledge of the NIR-II brain autofluorescence, responsi-ble for the loss of image resolution and contrast, is required. This work demon-strates that the brain shows a peculiar infrared autofluorescence spectrumthat can be correlated with specific molecular components. The existence ofparticular structures within the brain with well-defined NIR autofluorescencefingerprints is also evidenced, opening the door to in vivo anatomical imaging.Finally, we propose a rational selection of NIR luminescent probes suitable forlow-noise brain imaging based on their spectral overlap with brainautofluorescenceComunidad de Madrid, Grant/AwardNumber: B2017/BMD-3867RENIMCM;European Cooperation in Science andTechnology, Grant/Award Number:CA17140; Fundación para la Investigación Biomédica del Hospital Universitario Ramón y Cajal, Grant/Award Number:IMP18_38(2018/0265); Horizon 2020 Framework Programme, Grant/AwardNumber: 801305; Instituto de Salud CarlosIII, Grant/Award Number: PI16/00812;Ministerio de Ciencia, Innovación y Universidades, Grant/Award Number:FJC2018-036734-I; Ministerio deEconomía y Competitividad, Grant/AwardNumbers: MAT2016-75362-C3-1-R,MAT2017-83111R, MAT2017-85617-

In vivo grading of lipids in fatty liver by near-infrared autofluorescence and reflectance

The prevalence of nonalcoholic fatty liver (NAFLD) is rapidly increasing worldwide. When untreated, it may lead to complications such as liver cirrhosis or hepatocarcinoma. The diagnosis of NAFLD is usually obtained by ultrasonography, a technique that can underestimate its prevalence. For this reason, physicians aspire for an accurate, cost-effective, and noninvasive method to determine both the presence and the specific stage of the NAFLD. In this paper, we report an integrated approach for the quantitative estimation of the density of triglycerides in the liver based on the use of autofluorescence and reflectance signals generated by the abdomen of obese C57BL6/J mice. Singular value decomposition is applied to the generated spectra and its corresponding regression model provided a determination coefficient of 0.99 and a root mean square error of 240 mg/dl. This, in turn, enabled the quantitative imaging of triglycerides density in the livers of mice under in vivo conditionsMinisterio de Ciencia e Innovacion, Grant/Award Number: IJC2020-045229-I; Ministerio de Ciencia e Innovacion, Grant/Award Number: NANONERVPID2019-106211RB-I0

3D Optical Coherence Thermometry Using Polymeric Nanogels

In nanothermometry, the use of nanoparticles as thermal probes enables remote and minimally invasive sensing. In the biomedical context, nanothermometry has emerged as a powerful tool where traditional approaches, like infrared thermal sensing and contact thermometers, fall short. Despite the strides of this technology in preclinical settings, nanothermometry is not mature enough to be translated to the bedside. This is due to two major hurdles: the inability to perform 3D thermal imaging and the requirement for tools that are readily available in the clinics. This work simultaneously overcomes both limitations by proposing the technology of optical coherence thermometry (OCTh). This is achieved by combining thermoresponsive polymeric nanogels and optical coherence tomography (OCT)—a 3D imaging technology routinely used in clinical practice. The volume phase transition of the thermoresponsive nanogels causes marked changes in their refractive index, making them temperature-sensitive OCT contrast agents. The ability of OCTh to provide 3D thermal images is demonstrated in tissue phantoms subjected to photothermal processes, and its reliability is corroborated by comparing experimental results with numerical simulations. The results included in this work set credible foundations for the implementation of nanothermometry in the form of OCTh in clinical practiceThis work was financed by the Spanish Ministerio de Innovación y Ciencia under project NANONERV PID2019-106211RB-I00, NANOGRANZ PID2021-123318OB-I00, PID2020-118878RB-I00, RYC2021-032913-I, and TED2021-132317-I00B and under project COLUMNAS (PID2019- 110632RB-I00), by the Instituto de Salud Carlos III (PI19/00565), by the Comunidad Autónoma de Madrid (S2022/BMD-7403 RENIM-CM and SI3/PJI/2021-00211) and co-financed by the European structural and investment fund. Additional funding was provided by COST action CA17140, supported by COST (European Cooperation in Science and Technology) and the Fundación para la Investigación Biomédica del Hospital Universitario Ramón y Cajal (IMP21_A4 (2021/0427)

Early in vivo detection of denervation-induced atrophy by luminescence transient nanothermometry

Denervation induces skeletal muscle atrophy due to the loss of control and feedback with the nervous system. Unfortunately, muscle atrophy only becomes evident days after the denervation event when it could be irreversible. Alternative diagnosis tools for early detection of denervation-induced muscle atrophy are, thus, required. In this work, we demonstrate how the combination of transient thermometry, a technique already used for early diagnosis of tumors, and infrared-emitting nanothermometers makes possible the in vivo detection of the onset of muscle atrophy at short (<1 day) times after a denervation event. The physiological reasons behind these experimental results have been explored by performing three dimensional numerical simulations based on the Pennes' bioheat equation. It is concluded that the alterations in muscle thermal dynamics at the onset of muscle atrophy are consequence of the skin perfusion increment caused by the alteration of peripheral nervous autonomous system. This work demonstrates the potential of infrared luminescence thermometry for early detection of diseases of the nervous system opening the venue toward the development of new diagnosis toolsComunidad de Madrid, Grant/Award Number: S2017/BMD-3867 RENIM-CM; COST action CA17140 (Nano2Clinic); European Structural and Investment Fund and the Ministerio de Economía y Competitividad-MINECO, Grant/Award Number: PID2019-106211RB-I00; Juan de la Cierva scholarship, Grant/Award Number: IJC2020-045229-