Two-Photon Microscopy (TPM) and Fluorescence Lifetime Imaging Microscopy (FLIM) of Retinal Pigment Epithelium (RPE) of Mice In Vivo

2018 ◽  
pp. 73-88 ◽  
Author(s):  
Yoko Miura
Keyword(s):  
Retinal Pigment Epithelium ◽  
Fluorescence Lifetime ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Lifetime Imaging

scholarly journals Progressive dysmorphia of retinal pigment epithelium in age related macular degeneration revealed by fluorescence lifetime imaging

2021 ◽  
Author(s):  
Martin Hammer ◽  
Juliane Jakob-Girbig ◽  
Linda Schwanengel ◽  
Christine A. Curcio ◽  
Somar Hasan ◽  
...  
Keyword(s):  
Retinal Pigment Epithelium ◽  
Macular Degeneration ◽  
Fluorescence Lifetime ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Age Related Macular Degeneration ◽  
Fundus Photography ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Age Related

AbstractPurposeTo observe changes of the retinal pigment epithelium (RPE) on the transition from dysmorphia to atrophy in age related macular degeneration (AMD) by fluorescence lifetime imaging ophthalmoscopy (FLIO).MethodsMultimodal imaging including color fundus photography (CFP), optical coherence tomography (OCT), fundus autofluorescence (FAF) imaging, and FLIO was performed in 40 eyes of 37 patients with intermediate AMD and no evidence for geographic atrophy or macular neovascularization) (mean age: 74.2±7.0 years). Twenty-three eyes were followed for 28.3±18.3 months. Seven eyes had a second follow up after 46.6±9.0 months. Thickened RPE on OCT, hyperpigmentation on CFP, and migrated RPE, seen as hyperreflective foci (HRF) on OCT, were identified. Fluorescence lifetimes in two spectral channels (SSC: 500-560 nm, LSC: 560-720 nm) as well as emission spectrum intensity ratio (ESIR) of the lesions were measured by FLIO.ResultsAs hyperpigmented areas form and RPE migrates into the retina, FAF lifetimes lengthen and ESRI of RPE cells increase. Thickened RPE showed lifetimes of 256±49 ps (SSC) and 336±35 ps (LSC) and an ESIR of 0.552±0.079. For hyperpigmentation, these values were 317±68 ps (p<0.001), 377±56 ps (p<0.001), and 0.609±0.081 (p=0.001), respectively, and for HRF 337±79 ps (p<0.001), 414±50 ps (p<0.001), and 0.654±0.075 (p<0.001).ConclusionsIn the process of RPE degeneration, comprising different steps of dysmorphia, hyperpigmentation, and migration, lengthening of FAF lifetimes and a hypsochromic shift of emission spectra can be observed by FLIO. Thus, FLIO might provide early biomarkers for AMD progression and contribute to our understanding of RPE pathology.

scholarly journals Fluorescence Lifetime Imaging Ophthalmoscopy of the Retinal Pigment Epithelium During Wound Healing After Laser Irradiation

2019 ◽  
Vol 8 (5) ◽  
pp. 12
Author(s):  
Alessa Hutfilz ◽  
Svenja Rebecca Sonntag ◽  
Britta Lewke ◽  
Dirk Theisen-Kunde ◽  
Salvatore Grisanti ◽  
...  
Keyword(s):  
Wound Healing ◽  
Retinal Pigment Epithelium ◽  
Laser Irradiation ◽  
Fluorescence Lifetime ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

A molecular rotor sensor for detecting mitochondrial viscosity in apoptotic cells by two-photon fluorescence lifetime imaging

2020 ◽  
Vol 44 (26) ◽  
pp. 11342-11348 ◽  
Author(s):  
Ming-Xuan Hou ◽  
Liu-Yi Liu ◽  
Kang-Nan Wang ◽  
Xi-Juan Chao ◽  
Rong-Xue Liu ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Living Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Molecular Rotor ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

A two-photon fluorescent probe was developed for detecting mitochondrial viscosity during apoptosis of living cells by two-photon microscopy (TPM) and fluorescence lifetime imaging microscopy (FLIM) with good selectivity and highly biocompatible.

Visualization of Cutaneous Distribution of Minocycline of a Topical Gel in Human Facial Skin with Two-Photon Excited Fluorescence Lifetime Imaging Microscopy (FLIM) and Phasor Analysis

2017 ◽  
Vol 1 ◽  
pp. s57
Author(s):  
Sinyoung Jeong ◽  
Maiko Hermsmeier ◽  
Sam Osseiran ◽  
Akira Yamamoto ◽  
Usha Nagavarapu ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Facial Skin ◽  
Topical Gel ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Phasor Analysis

Abstract Not AvailableStudy supported by BioPharmX.

scholarly journals Coarse-grained modeling of mitochondrial metabolism enables subcellular flux inference from fluorescence lifetime imaging microscopy

2020 ◽  
Author(s):  
Xingbo Yang ◽  
Daniel J. Needleman
Keyword(s):  
Fluorescence Lifetime ◽  
Metabolic Flux ◽  
Living Cells ◽  
Coarse Grained ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Metabolic Fluxes ◽  
Subcellular Resolution

AbstractMitochondria are central to metabolism and their dysfunctions are associated with many diseases1–9. Metabolic flux, the rate of turnover of molecules through a metabolic pathway, is one of the most important quantities in metabolism, but it remains a challenge to measure spatiotemporal variations in mitochondrial metabolic fluxes in living cells. Fluorescence lifetime imaging microscopy (FLIM) of NADH is a label-free technique that is widely used to characterize the metabolic state of mitochondria in vivo10–18. However, the utility of this technique has been limited by the inability to relate FLIM measurement to the underlying metabolic activities in mitochondria. Here we show that, if properly interpreted, FLIM of NADH can be used to quantitatively measure the flux through a major mitochondrial metabolic pathway, the electron transport chain (ETC), in vivo with subcellular resolution. This result is based on the use of a coarse-grained NADH redox model, which we test in mouse oocytes subject to a wide variety of perturbations by comparing predicted fluxes to direct biochemical measurements and by self-consistency criterion. Using this method, we discovered a subcellular spatial gradient of mitochondrial metabolic flux in mouse oocytes. We showed that this subcellular variation in mitochondrial flux correlates with a corresponding subcellular variation in mitochondrial membrane potential. The developed model, and the resulting procedure for analyzing FLIM of NADH, are valid under nearly all circumstances of biological interest. Thus, this approach is a general procedure to measure metabolic fluxes dynamically in living cells, with subcellular resolution.

scholarly journals High-speed imaging of transient metabolic dynamics using two-photon fluorescence lifetime imaging microscopy

Optica ◽  
2018 ◽  
Vol 5 (10) ◽  
pp. 1290 ◽  
Author(s):  
Andrew J. Bower ◽  
Joanne Li ◽  
Eric J. Chaney ◽  
Marina Marjanovic ◽  
Darold R. Spillman ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
High Speed Imaging ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence
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