Rapid diagnosis and intraoperative margin assessment of human lung cancer with fluorescence lifetime imaging microscopy

Abstract

A method of rapidly differentiating lung tumor from healthy tissue is extraordinarily needed for both the diagnosis and the intraoperative margin assessment. We assessed the ability of fluorescence lifetime imaging microscopy (FLIM) for differentiating human lung cancer and normal tissues with the autofluorescence, and also elucidated the mechanism in tissue studies and cell studies. A 15-patient testing group was used to compare FLIM results with traditional histopathology diagnosis. Based on the endogenous fluorescence lifetimes of the testing group, a criterion line was proposed to distinguish normal and cancerous tissues. Then by blinded examined 41 sections from the validation group of other 16 patients, the sensitivity and specificity of FLIM were determined. The cellular metabolism was studied with specific perturbations of oxidative phosphorylation and glycolysis in cell studies. The fluorescence lifetime of cancerous lung tissues is consistently lower than normal tissues, and this is due to the both decrease of reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) lifetimes. A criterion line of lifetime at 1920 ps can be given for differentiating human lung cancer and normal tissues.The sensitivity and specificity of FLIM for lung cancer diagnosis were determined as 92.9% and 92.3%. These findings suggest that NADH and FAD can be used to rapidly diagnose lung cancer. FLIM is a rapid, accurate and highly sensitive technique in the judgment during lung cancer surgery and it can be potential in earlier cancer detection.

Molecular mobility and activity in an intravital imaging setting – implications for cancer progression and targeting

“…Fluorescence lifetime imaging microscopy (FLIM) Quantification of inherent fluorescence lifetime of a fluorophore as an exponential decay rate. The fluorescence lifetime is the time a fluorophore spends in the excited state before emitting a photon and returning to the ground state (Huck et al, 2016;Kalinina et al, 2016;Stringari et al, 2012;Walsh et al, 2013;Wang et al, 2017). Intramolecular FRET biosensors Donor and acceptor fluorophores with linker in between that can be altered by kinases, phosphatases, proteases or protein interaction (Fig.…”

“…Here, label-free fluorescence lifetime imaging microscopy (FLIM, see Box 1 for further details) readily lends itself to applications in humans. For instance, FLIM has been demonstrated in human lung and skin samples to delineate the border of cancer lesions (Galletly et al, 2008;Wang et al, 2017), as well as in capillaries of the fingernail bed and of the forearm in vivo (Shirshin et al, 2017) or in the tongue of human volunteers , which could also be adapted for the monitoring of oral cancers in humans. In a recent clinical trial, intravital tomography combined with FLIM of metabolites (see Table 1 and Box 1) demonstrated differences in mitochondrial density between healthy and inflamed skin and that an increase of free NADH in mitochondria correlated with the severity of inflammation (Huck et al, 2016).…”

Detecting benign uterine tumors by autofluorescence lifetime imaging microscopy through adjacent healthy cervical tissues

“…Optical imaging techniques especially°uorescence lifetime imaging microscopy (FLIM) are promising for the detection of malignancies [1][2][3][4][5][6] Di®erentiating malignancies from normal tissues by auto-°u orescence can rely on the biochemical and tissue morphological changes. The traditional gold standard technique for tissue characterization is haematoxylin and eosin (H & E) stained histopathology, based on morphological changes.…”

“…10,11 While NAD(P)H and FAD are known as classic molecules involving in oxidative phosphorylation and glycolysis, [12][13][14][15] which can be used to describe the cellular metabolic environment in tissues. Based on FLIM, di®erent cancers including cervical cancer, 4 lung cancer, 6 oral cancer, 16 breast cancer, 17 were studied on the auto°uorescence for early detection. Nevertheless, little attention was paid on benign tumors so far.…”

Ultra-fast fit-free analysis of complex fluorescence lifetime imaging via deep learning

“…Among all molecular imaging modalities, fluorescence optical imaging is a central technique thanks to its high sensitivity, the numerous molecular probes available, either endogenous or exogenous, and its ability to simultaneously image multiple biomarkers or biological processes at various spatio-temporal scales 1,2 . Especially, fluorescence lifetime imaging (FLI) has become an ever increasingly popular method as it provides unique insights into the cellular microenvironment, by non-invasively examining numerous intracellular parameters 3 such as metabolic status 4 , reactive oxygen species 5 and intracellular pH 6 . Moreover, FLI's exploitation of native fluorescent signatures has been extensively investigated for enhanced diagnostic of numerous pathologies.…”