Multimodal fluorescence molecular imaging for <i>in vivo</i> characterization of skin cancer using endogenous and exogenous fluorophores

Miller, J. Philip; Habimana-Griffin, LeMoyne; Edwards, Tracy S.; Achilefu, Samuel

doi:10.1117/1.jbo.22.6.066007

Cited by 15 publications

(8 citation statements)

References 29 publications

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“…[12], where a relationship was established between cellular autofluorescence and cellular metabolic processes, cell-endogenous fluorophores have been successfully used as biomarkers in the non-destructive and real-time determination of cell characteristics. Numerous adoptions have thus been made in biomedical research and diagnosis [13], with notable applications in the identification of stem cell differentiation [14, 15] as well as the detection of diseases such as cancer [16, 17] and Alzheimer’s [18]. These applications have been enabled by optical techniques such as multi-photon microscopy cum spectroscopy [15, 19] and fluorescence lifetime imaging microscopy [20, 21].…”

Section: Introductionmentioning

confidence: 99%

Autofluorescence spectroscopy in redox monitoring across cell confluencies

et al. 2019

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Patient-specific therapies require that cells be manufactured in multiple batches of small volumes, making it a challenge for conventional modes of quality control. The added complexity of inherent variability (even within batches) necessitates constant monitoring to ensure comparable end products. Hence, it is critical that new non-destructive modalities of cell monitoring be developed. Here, we study, for the first time, the use of optical spectroscopy in the determination of cellular redox across cell confluencies by exploiting the autofluorescence properties of molecules found natively within cells. This was achieved through a simple retrofitting of a standard inverted fluorescence microscope with a spectrometer output and an appropriate fluorescence filter cube. Through spectral decomposition on the acquired autofluorescence spectra, we are able to further discern the relative contributions of the different molecules, namely flavin adenine dinucleotide (FAD) and reduced nicotinamide adenine dinucleotide (NADH). This is then quantifiable as redox ratios (RR) that represent the extent of oxidation to reduction based upon the optically measured quantities of FAD and NADH. Results show that RR decreases with increasing cell confluency, which we attribute to several inter-related cellular processes. We validated the relationship between RR, metabolism and cell confluency through bio-chemical and viability assays. Live-dead and DNA damage studies were further conducted to substantiate that our measurement process had negligible effects on the cells. In this study, we demonstrate that autofluorescence spectroscopy-derived RR can serve as a rapid, non-destructive and label-free surrogate to cell metabolism measurements. This was further used to establish a relationship between cell metabolism and cellular redox across cell confluencies, and could potentially be employed as an indicator of quality in cell therapy manufacturing.

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Section: Introductionmentioning

confidence: 99%

Autofluorescence spectroscopy in redox monitoring across cell confluencies

et al. 2019

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“…In addition, SCC-74A and SCC-74B could be clearly distinguished based on the dynamic range of°u orescence intensity (the di®erence between uncoupled and inhibited metabolic states) and the lifetime distribution. In 2017, Miller et al 42 characterized skin cancer in living mice with SCC by using multimodal°uorescence molecular imaging method. Firstly, the endogenous°uorophores, such as°avin and lipofuscin, were excited by 480 nm laser.…”

Section: Squamous Cell Carcinomamentioning

confidence: 99%

Fluorescence lifetime imaging microscopy and its applications in skin cancer diagnosis

Liu

Yang

Zhang

et al. 2019

J. Innov. Opt. Health Sci.

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Fluorescence lifetime (FLT) of fluorophores is sensitive to the changes in their surrounding microenvironment, and hence it can quantitatively reveal the physiological characterization of the tissue under investigation. Fluorescence lifetime imaging microscopy (FLIM) provides not only morphological but also functional information of the tissue by producing spatially resolved image of fluorophore lifetime, which can be used as a signature of disorder and/or malignancy in diseased tissues. In this paper, we begin by introducing the basic principle and common detection methods of FLIM. Then the recent advances in the FLIM-based diagnosis of three different skin cancers, including basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and malignant melanoma (MM) are reviewed. Furthermore, the potential advantages of FLIM in skin cancer diagnosis and the challenges that may be faced in the future are prospected.

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“…Since the pioneering work by Chance et al, 8 where a relationship was established between cell autofluorescence and cellular metabolic processes, cell-endogenous fluorophores have been successfully used as biomarkers in the nondestructive and real-time determination of cell characteristics. Numerous adoptions have thus been made in biomedical research and diagnosis, 9 with notable applications in the identification of stem cell differentiation 10,11 as well as the detection of diseases such as cancer 12,13 and Alzheimer's. 14 These applications have been enabled by optical techniques such as multi-photon microscopy cum spectroscopy 11,15 and fluorescence lifetime imaging microscopy.…”

Section: Introductionmentioning

confidence: 99%

Autofluorescence spectroscopy for cell monitoring

Yong

Rahim

Ong

et al. 2018

CLEO Pacific Rim Conference

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Patient-specific therapies require that cells be manufactured in multiple batches of small volumes, making it a challenge for conventional modes of quality control. The added complexity of inherent variability (even within batches) necessitates constant monitoring to ensure comparable end products. Hence, it is critical that new non-destructive modalities of cell monitoring be developed. Here, we study, for the first time, the use of optical spectroscopy in the determination of cell confluency. We exploit the autofluorescence properties of molecules found natively within cells. By applying spectral decomposition on the acquired autofluorescence spectra, we are able to further discern the relative contributions of the different molecules, namely flavin adenine dinucleotide (FAD) and reduced nicotinamide adenine dinucleotide (NADH). This is then quantifiable as redox ratios that represent the extent of oxidation to reduction based upon the optically measured quantities of FAD and NADH. Results show that RR is significantly higher for lower confluencies (≤50%), which we attribute to the different metabolic requirements as cells switch from individual survival to concerted proliferation. We validate this relationship through bio-chemical assays and autofluorescence imaging, and further confirmed through a live-dead study that our measurement process had negligible effects on cell viability.

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Multimodal fluorescence molecular imaging for in vivo characterization of skin cancer using endogenous and exogenous fluorophores

Cited by 15 publications

References 29 publications

Autofluorescence spectroscopy in redox monitoring across cell confluencies

Autofluorescence spectroscopy in redox monitoring across cell confluencies

Fluorescence lifetime imaging microscopy and its applications in skin cancer diagnosis

Autofluorescence spectroscopy for cell monitoring

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