A5083395619- Advanced Fluorescence Microscopy Techniques
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- 3D Printing in Biomedical Research
- Cancer, Hypoxia, and Metabolism
- Spectroscopy Techniques in Biomedical and Chemical Research
- Immune cells in cancer
- Neuroscience and Neural Engineering
- Photoacoustic and Ultrasonic Imaging
- Nanoplatforms for cancer theranostics
- Immune Response and Inflammation
- Single-cell and spatial transcriptomics
- Adipose Tissue and Metabolism
- S100 Proteins and Annexins
- Cell Image Analysis Techniques
- Cancer Research and Treatments
- Blood disorders and treatments
- Metabolomics and Mass Spectrometry Studies
- Analytical Chemistry and Sensors
- Cell Adhesion Molecules Research
- Pluripotent Stem Cells Research
- bioluminescence and chemiluminescence research
- Birth, Development, and Health
- Genetics, Aging, and Longevity in Model Organisms
- Cardiac electrophysiology and arrhythmias
- Agriculture, Soil, Plant Science
Morgridge Institute for Research
2018-2025
University of California, Irvine
2014-2025
Institute of Biomedical Science
2024
Zero to Three
2021
Fluor (United States)
2014-2019
Jadavpur University
2010
Abstract There is a growing interest in developing microphysiological systems that can be used to model both normal and pathological human organs vitro . This “organs-on-chips” approach aims capture key structural physiological characteristics of the target tissue. Here we describe vascularized microtumors (VMTs). “tumor-on-a-chip” platform incorporates tumor stromal cells grow 3D extracellular matrix depend for survival on nutrient delivery through living, perfused microvessels. Both...
Conventionally, the size, shape, and biomechanics of cartilages are determined by their voluminous extracellular matrix. By contrast, we found that multiple murine consist lipid-filled cells called lipochondrocytes. Despite resembling adipocytes, lipochondrocytes were molecularly distinct produced lipids exclusively through de novo lipogenesis. Consequently, grew uniform lipid droplets resisted systemic surges did not enlarge upon obesity. Lipochondrocytes also lacked mobilization factors,...
Abstract Presence of reactive oxygen species (ROS) in excess normal physiological level results oxidative stress. This can lead to a range pathological conditions including inflammation, diabetes mellitus, cancer, cardiovascular and neurodegenerative disease. Biomarkers stress play an important role understanding the pathogenesis treatment these diseases. A number fluorescent biomarkers exist. However, non-invasive label-free identification technique would be advantageous for vivo...
The majority of cancer cells have a unique metabolic requirement for methionine that is not observed in normal, non-tumorigenic cells. This phenotype described as "methionine dependence" or stress sensitivity" which are unable to proliferate when has been replaced with its precursor, homocysteine, cell culture growth media. We focus on the response triple negative breast line MDA-MB-468 and insensitive derivative MDA-MB-468res-R8. Using variety techniques including fluorescence lifetime...
Macrophages adopt a variety of phenotypes that are reflection the many functions they perform as part immune system. In particular, metabolism is phenotypic trait differs between classically activated, proinflammatory macrophages, and alternatively prohealing macrophages. Inflammatory macrophages have based on glycolysis while activated generally rely oxidative phosphorylation to generate chemical energy. We employ this shift in an endogenous marker identify phenotype individual via...
Significance: Fluorescence lifetime imaging microscopy (FLIM) measures the decay rate of fluorophores, thus providing insights into molecular interactions. FLIM is a powerful technique that widely used in biology and medicine. Aim: This perspective highlights some major advances instrumentation, analysis, biological clinical applications we have found impactful over last year. Approach: Innovations instrumentation resulted faster acquisition speeds, rapid large fields view, integration with...
Abstract Bacterial populations exhibit a range of metabolic states influenced by their environment, intra- and interspecies interactions. The identification bacterial transitions between them in native environment promises to elucidate community behavior stochastic processes, such as antibiotic resistance acquisition. In this work, we employ two-photon fluorescence lifetime imaging microscopy (FLIM) create fingerprint individual bacteria populations. FLIM autofluorescent reduced nicotinamide...
Access to electron acceptors supports oxidized biomass synthesis and can be limiting for cancer cell proliferation, but how cells overcome this limitation in tumors is incompletely understood. Nontransformed help metabolic limitations, particularly pancreatic cancer, where stellate (PSCs) promote proliferation tumor growth. However, whether PSCs affect the redox state of not known. By taking advantage endogenous fluorescence properties reduced nicotinamide adenine dinucleotide flavin...
Neutrophils, the most abundant leukocytes in human peripheral circulation, are crucial for innate immune response. They typically quiescent but rapidly activate response to infection and inflammation, performing diverse functions such as oxidative burst, phagocytosis, NETosis, which require significant metabolic adaptation. Deeper insights into changes will help identify regulation of neutrophil health diseases. Due their short lifespan associated technical challenges, processes neutrophils...
Neutrophils are critical for host defense against fungi. However, the short life span and lack of genetic tractability primary human neutrophils has limited in vitro analysis neutrophil-fungal interactions. Human induced pluripotent stem cell (iPSC)-derived (iNeutrophils) provide a genetically tractable system to study responses neutrophils. Here, we show that deletion transcription factor GATA1 from iPSCs results iNeutrophils with improved antifungal activity Aspergillus fumigatus....
Abstract Neutrophils are highly abundant innate immune cells that constantly produced from myeloid progenitors in the bone marrow. Differentiated neutrophils can perform an arsenal of effector functions critical for host defense. This study aims to quantitatively understand neutrophil mitochondrial metabolism throughout differentiation and activation, elucidate impact on functions. To metabolic remodeling differentiation, murine ER-Hoxb8 progenitor-derived human induced pluripotent stem...
In this work we demonstrate a label-free optical imaging technique to assess metabolic status and oxidative stress in human induced pluripotent stem cell-derived cardiomyocytes by two-photon fluorescence lifetime of endogenous fluorophores. Our results show the sensitivity method detect shifts metabolism upon pathological stimuli hypoxia cardiotoxic drugs. This non-invasive could prove beneficial for drug development screening, especially vitro cardiac models created from study pathogenesis...
Phasor FLIM in cells undergoing oxidative stress and mice liver sections have shown the presence of a third autofluorescent component indicative lipid droplets along with free enzyme-bound NADH similar emissions. This affects position shape phasor distribution, pushing it away from metabolic trajectory. rule addition is still valid was exploited here to create multicomponent analysis where distribution can be reassigned trajectory changes metabolism detected independently intensity this...
Supervised machine learning can be used to predict which drugs human cardiomyocytes have been exposed to. Using electrophysiological data collected from with known exposure different drugs, a supervised algorithm trained recognize and classify cells that an unknown drug. Furthermore, the provides information on relative contribution of each parameter overall classification. Probabilities confidence in accuracy classification may also determined by algorithm. In this study, effects...
There is an ongoing need to do more with less and provide highly multiplexed analysis from limited sample volumes. Improved "sample sparing" assays would have a broad impact across pediatric other rare type studies in addition enabling sequential sampling. This capability advance both clinical basic research applications. Here we report the micro blood technology (μBAT), microfluidic platform that supports of neutrophils single drop blood. We demonstrate orthogonal capabilities μBAT...
Abstract Neutrophils are critical for host defense against fungi. However, the short life span and lack of genetic tractability primary human neutrophils has limited in vitro analysis neutrophil-fungal interactions. Human induced pluripotent stem cell (iPSC)-derived (iNeutrophils) a genetically tractable alternative to neutrophils. Here, we show that deletion transcription factor GATA1 from iPSCs results iNeutrophils with improved antifungal activity Aspergillus fumigatus . knockout (KO)...
SignificanceAdvancements in label-free microscopy could provide real-time, non-invasive imaging with unique sources of contrast and automated standardized analysis to characterize heterogeneous dynamic biological processes. These tools would overcome challenges widely used methods that are destructive (e.g., histology, flow cytometry) or lack cellular resolution plate-based assays, whole animal bioluminescence imaging).AimThis perspective aims (1) justify the need for track functions over...
Significance: Glioblastoma multiforme (GBM) is the most frequently diagnosed adult primary brain malignancy with poor patient prognosis. GBM can recur despite aggressive treatment due to therapeutically resistant glioblastoma stem cells (GSCs) that may exhibit metabolic plasticity. Aim: Intrinsic nicotinamide adenine dinucleotide (NADH) fluorescence be acquired lifetime imaging microscopy (FLIM) examine its bound and free states in GSC tissues. Approach: We compared mean NADH live human GSCs...
Metabolic features of mitosis remain poorly understood because this phase the cell cycle is rapid and heterogeneous between cells within a dish. Label-free optical metabolic imaging (OMI) can monitor changes in metabolism with single resolution using two-photon microscopy redox ratio (NAD(P)H/FAD) NAD(P)H fluorescence lifetimes. Here, we brought together image analysis tools to quantify OMI time-courses undergoing across multiple lines. Alignment morphological over time provided unique...