Abnormal cellular metabolism is a hallmark of cancer, yet there an absence quantitative methods to dynamically image this powerful function. Optical metabolic imaging (OMI) noninvasive, high-resolution, tool for monitoring metabolism. OMI probes the fluorescence intensities and lifetimes autofluorescent coenzymes reduced NADH flavin adenine dinucleotide. We confirm that correlates with glycolytic levels across panel human breast cell lines using standard assays rates glucose uptake lactate...

Abstract There is a need for technologies to predict the efficacy of cancer treatment in individual patients. Here, we show that optical metabolic imaging organoids derived from primary tumors can therapeutic response xenografts and measure antitumor drug responses human tumor–derived organoids. Optical quantifies fluorescence intensity lifetime NADH FAD, coenzymes metabolism. As early as 24 hours after with clinically relevant anticancer drugs, index responsive decreased (P < 0.001)...

Objectives Three-dimensional organoids derived from primary pancreatic ductal adenocarcinomas are an attractive platform for testing potential anticancer drugs on patient-specific tissue. Optical metabolic imaging (OMI) is a novel tool used to assess drug-induced changes in cellular metabolism, and its quantitative end point, the OMI index, evaluated as biomarker of drug response cancer organoids. Methods both malignant cell fibroblast within murine human Results Anticancer induce...

Subpopulations of cells that escape anti-cancer treatment can cause relapse in cancer patients. Therefore, measurements cellular-level tumor heterogeneity could enable improved regimens. Cancer exhibits altered cellular metabolism, which affects the autofluorescence metabolic cofactors NAD(P)H and FAD. The optical redox ratio (fluorescence intensity divided by FAD) reflects global metabolism. fluorescence lifetime (amount time a fluorophore is excited state) sensitive to microenvironment,...

The genetic and phenotypic heterogeneity of cancers can contribute to tumor aggressiveness, invasion, resistance therapy.Fluorescence imaging occupies a unique niche investigate due its high resolution molecular specificity.Here, heterogeneous populations are identified quantified by combined optical metabolic subpopulation analysis (OMI-SPA).OMI probes the fluorescence intensities lifetimes enzymes in cells provide images cellular metabolism, SPA models cell as mixed Gaussian distributions...

Abstract Primary tumor organoids grown in three-dimensional culture provide an excellent platform for studying progression, invasion and drug response. However, organoid generation protocols require fresh tissue, which limits research clinical use. This study investigates cellular morphology, viability response of derived from frozen tissues. The results demonstrate that viable can be flash-frozen thawed tissue bulk tissues slowly DMSO supplemented media. While the freezing process affects...

The function of macrophages in vitro is linked to their metabolic rewiring. However, macrophage metabolism remains poorly characterized situ. Here, we used two-photon intensity and lifetime imaging autofluorescent coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) flavin (FAD), assess the wound microenvironment. Inhibiting glycolysis reduced NAD(P)H mean made intracellular redox state more oxidized, as indicated by optical ratio. We found that TNFα+ had lower were oxidized...

Optical metabolic imaging measures fluorescence intensity and lifetimes from cofactors nicotinamide adenine dinucleotide (NADH) flavin (FAD). These molecular level measurements provide unique biomarkers for early cellular responses to cancer treatments. Head neck squamous cell carcinoma (HNSCC) is an attractive target optical because of easy access the site using fiber optic probes. Two HNSCC lines, SCC25 SCC61, were treated with Cetuximab (anti-EGFR antibody), BGT226 (PI3K/mTOR inhibitor),...

The optical redox ratio (fluorescence intensity of NADH divided by that FAD), was acquired for a panel breast cancer cell lines to investigate how overexpression human epidermal growth factor receptor 2 (HER2) affects tumor metabolism, and metabolism may be altered in response clinically used HER2-targeted therapies. Confocal fluorescence microscopy acquire FAD auto-fluorescent images. highest cells overexpressing HER2 lowest triple negative (TNBC) cells, which lack HER2, progesterone...

Light-sheet microscopy is an increasingly popular imaging tool for studying biological processes, however, it generally requires the use of exogenous fluorescent contrast agents which can be toxic to live cells. tomography label-free analog light-sheet microscopy, utilizing elastic scattering visualize structure sample. Here, we present a multi-modal polarization and (PLμTo) system on-line, non-destructive, non-invasive volumetric human mesenchymal stem cells, also referred as stromal...

The polymerase gamma (POLG) gene mutation is associated with mitochondria and metabolism disorders, resulting in heterogeneous responses to immunological activation posing challenges for mitochondrial disease therapy. Optical metabolic imaging captures the autofluorescent signal of two coenzymes, NADH FAD, offers a label-free approach detect cellular phenotypes, track morphology, quantify heterogeneity. In this study, fluorescence lifetime (FLIM) NAD(P)H FAD revealed that POLG mutator...

Cryopreservation is a widely used technique to preserve biological samples for extended periods of time at low temperatures. Even though it known have significant effects on cell viability, its effect their metabolism remains unexplored. Studying how cryopreservation influences the cells important guarantee reliability transported between sites analysis. Optical metabolic imaging allows study cellular in label-free manner by using autofluorescence properties nicotinamide adenine dinucleotide...

Luminescence lifetime imaging reports information about the local environment of luminescent molecules. Traditional systems for luminescence typically rely on ultrafast detectors and electronics. While these offer high temporal resolution, their cost complexity limit widespread applicability. To address this gap, a novel optical system, MAX-alif (Modular Analysis eXtraction system Affordable Lifetime Imaging Fluorescence signals), was developed to provide speed, low-cost scalable solution...

Luminescence lifetime imaging reports information about the local environment of luminescent molecules. Traditional systems for luminescence typically rely on ultrafast detectors and electronics. While these offer high temporal resolution, their cost complexity limit widespread applicability. To address this gap, a novel optical system, MAX-alif (Modular Analysis eXtraction system Affordable Lifetime Imaging Fluorescence signals), was developed to provide speed, low-cost scalable solution...

Abstract Triple-negative breast cancer (TNBC) is an aggressive subtype of with no targeted treatments currently available. TNBC cells participate in metabolic symbiosis, a process that optimizes tumor growth by balancing processes between glycolysis and oxidative phosphorylation through increased activity the enzyme lactate dehydrogenase B (LDHB). Metabolic symbiosis allows to function at similar rate as glycolytic cells, increasing overall proliferation. Here, fluorescence lifetime imaging...

Mitochondria are dynamic organelles that play a key role in energy production and maintaining cellular homeostasis. The regulation of mitochondrial dynamics, involving both fission fusion, is vital for healthy population mitochondria within the cell. Alterations dynamics have been associated with various disease states, such as metabolic neurodegenerative diseases cancer. We describe protocol imaging analyzing NAD(P)H intensity to visualize movement over time 3D distribution network A...

The mechanisms of many anesthetic drugs, such as propofol, are unknown. This study investigates the impact a widely used intravenous drug, on metabolic behavior human triple-negative breast cancer cell line, MDA-MB 231. Utilizing fluorescence lifetime imaging microscopy (FLIM), we assessed effects propofol cellular metabolism through lifetimes NADH and FAD, coenzymes reactions. Exposure to induced significant morphological changes cells, including substantial shrinkage, which may reveal...