Phasor analysis of multiphoton spectral images distinguishes autofluorescence components of <i>in vivo</i> human skin

Fereidouni, Farzad; Bader, Arjen N.; Colonna, Anne; Gerritsen, Hans C.

doi:10.1002/jbio.201200244

Cited by 59 publications

(64 citation statements)

References 25 publications

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“…By analyzing the spectroscopic properties of the emission -e.g. by using fluorescence lifetime imaging (FLIM) or spectral imaging -multiple signals originating from, for example, NADH, flavins, retinol, keratin, FAD, melanin and elastin can be separated and quantified (Fereidouni et al, 2014;Skala et al, 2007;Zipfel et al, 2003). Compared to THG, autofluorescence signals require shorter near infrared excitation wavelengths and usually have a low signal-to-noise ratio, which limits detection in scattering tissues.…”

Section: Box 1 Label-free Imaging Modalities In Life Sciencesmentioning

confidence: 99%

Third harmonic generation microscopy of cells and tissue organization

Weigelin

Bakker

Friedl

2016

Journal of Cell Science

177

147

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The interaction of cells within their microenvironmental niche is fundamental to cell migration, positioning, growth and differentiation in order to form and maintain complex tissue organization and function. Third harmonic generation (THG) microscopy is a label-free scatter process that is elicited by water-lipid and water-protein interfaces, including intra-and extracellular membranes, and extracellular matrix structures. In applied life sciences, THG delivers a versatile contrast modality to complement multi-parameter fluorescence, second harmonic generation and fluorescence lifetime microscopy, which allows detection of cellular and molecular cell functions in threedimensional tissue culture and small animals. In this Commentary, we review the physical and technical basis of THG, and provide considerations for optimal excitation, detection and interpretation of THG signals. We further provide an overview on how THG has versatile applications in cell and tissue research, with a particular focus on analyzing tissue morphogenesis and homeostasis, immune cell function and cancer research, as well as the emerging applicability of THG in clinical practice.

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Section: Box 1 Label-free Imaging Modalities In Life Sciencesmentioning

confidence: 99%

Third harmonic generation microscopy of cells and tissue organization

Weigelin

Bakker

Friedl

2016

Journal of Cell Science

177

147

View full text Add to dashboard Cite

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“…phasor analysis [31, 32]) is expected to not improve the overall speed of displaying the augmented video frames. Due to parallelization of the processes the overall speed is determined by the formation of the FLIm maps (Table I).…”

Section: Resultsmentioning

confidence: 99%

Real-Time Visualization of Tissue Surface Biochemical Features Derived From Fluorescence Lifetime Measurements

Gorpas

Bec

et al. 2016

IEEE Trans. Med. Imaging

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Fiber based fluorescence lifetime imaging has shown great potential for intraoperative diagnosis and guidance of surgical procedures. Here we describe a novel method addressing a significant challenge for the practical implementation of this technique, i.e. the real-time display of the quantified biochemical or functional tissue properties superimposed on the interrogated area. Specifically, an aiming beam (450 nm) generated by a continuous-wave laser beam was merged with the pulsed fluorescence excitation light in a single delivery/collection fiber and then imaged and segmented using a color-based algorithm. We demonstrate that this approach enables continuous delineation of the interrogated location and dynamic augmentation of the acquired frames with the corresponding fluorescence decay parameters. The method was evaluated on a fluorescence phantom and fresh tissue samples. Current results demonstrate that 34 frames per second can be achieved for augmenting videos of 640×512 pixels resolution. Also we show that the spatial resolution of the fluorescence lifetime map depends on the tissue optical properties, the scanning speed, and the frame rate. The dice similarity coefficient between the fluorescence phantom and the reconstructed maps was estimated to be as high as 93%. The reported method could become a valuable tool for augmenting the surgeon’s field of view with diagnostic information derived from the analysis of fluorescence lifetime data in real-time using handheld, automated, or endoscopic scanning systems. Current method provides also a means for maintaining the tissue light exposure within safety limits. This study provides a framework for using an aiming beam with other point spectroscopy applications.

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“…Spectral phasor analysis was carried out using a custom ImageJ plugin (available from http://www.spechron.com). The phasor approach has previously been implemented to analyse lifetime [39, 40] and spectral images [41, 42]. A detailed description of this analysis is provided in the appendix.…”

Section: Methodsmentioning

confidence: 99%

Early BAFF receptor blockade mitigates murine Sjögren's syndrome: Concomitant targeting of CXCL13 and the BAFF receptor prevents salivary hypofunction

Sharma

Kiripolsky

Klimatcheva

et al. 2016

Clinical Immunology

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Sjögren's Syndrome (SS) is a debilitating autoimmune disease. Patients with SS may develop xerostomia. This process is progressive, and there are no therapeutics that target disease etiology. We hypothesized BAFF receptor (BAFFR) blockade would mitigate SS disease development, and neutralization of CXCL13 and BAFF signaling would be more efficacious than BAFFR blockade alone. We treated NOD/ShiLtJ SS mice with soluble BAFF receptor (BAFFR-Fc) or anti-CXCL13/BAFFR-Fc in combination, prior to the development of clinical disease. Our results show treatment with BAFFR-Fc reduced peripheral B cells numbers and decreased sialadenitis. In addition, this treatment reduced total serum immunoglobulin as well as IgG and IgM specific anti-nuclear autoantibodies. NOD/ShiLtJ mice treated with BAFFR-Fc and anti-CXCL13 antibody were protected from salivary deficits. Results from this study suggest blockade of CXCL13 and BAFFR together may be an effective therapeutic strategy in preventing salivary hypofunction and reducing autoantibody titers and sialadenitis in patients with SS.

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Phasor analysis of multiphoton spectral images distinguishes autofluorescence components of in vivo human skin

Cited by 59 publications

References 25 publications

Third harmonic generation microscopy of cells and tissue organization

Third harmonic generation microscopy of cells and tissue organization

Real-Time Visualization of Tissue Surface Biochemical Features Derived From Fluorescence Lifetime Measurements

Early BAFF receptor blockade mitigates murine Sjögren's syndrome: Concomitant targeting of CXCL13 and the BAFF receptor prevents salivary hypofunction

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