Label-Free Metabolic Classification of Single Cells in Droplets Using the Phasor Approach to Fluorescence Lifetime Imaging Microscopy.

Characterization of single cell metabolism is imperative for understanding subcellular functional and biochemical changes associated with healthy tissue development and the progression of numerous diseases. However, single-cell analysis often requires the use of fluorescent tags and cell lysis followed by genomic profiling to identify the cellular heterogeneity. Identifying individual cells in a noninvasive and label-free manner is crucial for the detection of energy metabolism which will discriminate cell types and most importantly critical for maintaining cell viability for further analysis. Here, we have developed a robust assay using the droplet microfluidic technology together with the phasor approach to fluorescence lifetime imaging microscopy to study cell heterogeneity within and among the leukemia cell lines (K-562 and Jurkat). We have extended these techniques to characterize metabolic differences between proliferating and quiescent cells-a critical step toward label-free single cancer cell dormancy research. The result suggests a droplet-based noninvasive and label-free method to distinguish individual cells based on their metabolic states, which could be used as an upstream phenotypic platform to correlate with genomic statistics. © 2018 International Society for Advancement of Cytometry

Blended Learning: Integrating an e-Learning Component for Competency Validation

Introduction: Emergency care providers are required to demonstrate competency in the management of life-threatening situation. The care provider’s ability to manage an emergency situation depends upon his/her knowledge and skills in basic CPR; and the use of emergency equipment and supplies. The education department at our healthcare facility is responsible for CPR/Emergency Management competency validation of over 2500 employees annually. Historically each employee was scheduled to attend 4 hours of class every year to review the content, complete the post-test and demonstrate skills. It was resource-intensive, time consuming, stressful and often difficult to schedule the 24/7 employees for the sessions. [See PDF for complete abstract

Rapid and label-free identification of single leukemia cells from blood in a high-density microfluidic trapping array by fluorescence lifetime imaging microscopy.

The rapid screening and isolation of single leukemia cells from blood has become critical for early leukemia detection and tumor heterogeneity interrogation. However, due to the size overlap between leukemia cells and the more abundant white blood cells (WBCs), the isolation and identification of leukemia cells individually from peripheral blood is extremely challenging and often requires immunolabeling or cytogenetic assays. Here we present a rapid and label-free single leukemia cell identification platform that combines: (1) high-throughput size-based separation of hemocytes via a single-cell trapping array, and (2) leukemia cell identification through phasor approach and fluorescence lifetime imaging microscopy (phasor-FLIM), to quantify changes between free/bound nicotinamide adenine dinucleotide (NADH) as an indirect measurement of metabolic alteration in living cells. The microfluidic trapping array designed with 1600 highly-packed addressable single-cell traps can simultaneously filter out red blood cells (RBCs) and trap WBCs/leukemia cells, and is compatible with low-magnification imaging and fast-speed fluorescence screening. The trapped single leukemia cells, e.g., THP-1, Jurkat and K562 cells, are distinguished from WBCs in the phasor-FLIM lifetime map, as they exhibit significant shift towards shorter fluorescence lifetime and a higher ratio of free/bound NADH compared to WBCs, because of their glycolysis-dominant metabolism for rapid proliferation. Based on a multiparametric scheme comparing the eight parameter-spectra of the phasor-FLIM signatures, spiked leukemia cells are quantitatively distinguished from normal WBCs with an area-under-the-curve (AUC) value of 1.00. Different leukemia cell lines are also quantitatively distinguished from each other with AUC values higher than 0.95, demonstrating high sensitivity and specificity for single cell analysis. The presented platform is the first to enable high-density size-based single-cell trapping simultaneously with RBC filtering and rapid label-free individual-leukemia-cell screening through non-invasive metabolic imaging. Compared to conventional biomolecular diagnostics techniques, phasor-FLIM based single-cell screening is label-free, cell-friendly, robust, and has the potential to screen blood in clinical volumes through parallelization

Properties and Performance of Two Wide Field of View Cherenkov/Fluorescence Telescope Array Prototypes

A wide field of view Cherenkov/fluorescence telescope array is one of the main components of the Large High Altitude Air Shower Observatory project. To serve as Cherenkov and fluorescence detectors, a flexible and mobile design is adopted for easy reconfiguring of the telescope array. Two prototype telescopes have been constructed and successfully run at the site of the ARGO-YBJ experiment in Tibet. The features and performance of the telescopes are presented

Critical Exponents of the Metal-Insulator Transition in the Two-Dimensional Hubbard Model

We study the filling-controlled metal-insulator transition in the two-dimensional Hubbard model near half-filling with the use of zero temperature quantum Monte Carlo methods. In the metallic phase, the compressibility behaves as $\kappa \propto |\mu - \mu_c|^{-0.58\pm0.08}$ where $\mu_c$ is the critical chemical potential. In the insulating phase, the localization length follows $\xi_l \propto |\mu - \mu_c|^{-\nu_l}$ with $\nu_l = 0.26 \pm 0.05$. Under the assumption of hyperscaling, the compressibility data leads to a correlation length exponent $\nu_\kappa = 0.21 \pm 0.04$. Our results show that the exponents $\nu_\kappa$ and $\nu_l$ agree within statistical uncertainty. This confirms the assumption of hyperscaling with correlation length exponent $\nu = 1/4$ and dynamical exponent $z = 4$. In contrast the metal-insulator transition in the generic band insulators in all dimensions as well as in the one-dimensional Hubbard model satisfy the hyperscaling assumption with exponents $\nu = 1/2$ and $z = 2$.Comment: Two references added. The DVI file and PS figure files are also available at http://www.issp.u-tokyo.ac.jp/labs/riron/imada/furukawa/; to appear in J. Phys. Soc. Jpn 65 (1996) No.

Dificultades en la clasificación del síndrome metabólico: El ejemplo de los adolescentes en México

OBJETIVO. Determinar la diferencia entre las definiciones del National Cholesterol Education Program Adult Treatment Panel (ATPIII) y de la International Diabetes Federation (IDF) para síndrome metabólico (SM) en adolescentes mexicanos. MATERIAL Y MÉTODOS. Estudio transversal en 575 adolescentes de 14 a 16 años. Se utilizaron pruebas t de Student, ji cuadrada y correlación de Spearman. RESULTADOS. La prevalencia de SM fue mayor por ATPIII (18.6%) versus IDF (8.2%) (p<0.001), con 41.1% de concordancia. CONCLUSIONES. Existe una diferencia estadística de la prevalencia del SM en adolescentes mexicanos entre las dos definiciones

Generalizing the O(N)-field theory to N-colored manifolds of arbitrary internal dimension D

We introduce a geometric generalization of the O(N)-field theory that describes N-colored membranes with arbitrary dimension D. As the O(N)-model reduces in the limit N->0 to self-avoiding polymers, the N-colored manifold model leads to self-avoiding tethered membranes. In the other limit, for inner dimension D->1, the manifold model reduces to the O(N)-field theory. We analyze the scaling properties of the model at criticality by a one-loop perturbative renormalization group analysis around an upper critical line. The freedom to optimize with respect to the expansion point on this line allows us to obtain the exponent \nu of standard field theory to much better precision that the usual 1-loop calculations. Some other field theoretical techniques, such as the large N limit and Hartree approximation, can also be applied to this model. By comparison of low and high temperature expansions, we arrive at a conjecture for the nature of droplets dominating the 3d-Ising model at criticality, which is satisfied by our numerical results. We can also construct an appropriate generalization that describes cubic anisotropy, by adding an interaction between manifolds of the same color. The two parameter space includes a variety of new phases and fixed points, some with Ising criticality, enabling us to extract a remarkably precise value of 0.6315 for the exponent \nu in d=3. A particular limit of the model with cubic anisotropy corresponds to the random bond Ising problem; unlike the field theory formulation, we find a fixed point describing this system at 1-loop order.Comment: 57 pages latex, 26 figures included in the tex

Stellar Population Properties and Evolution Analysis of NGC 628 with the Panchromatic Photometry

Panchromatic spectral energy distribution (SED) from the ultraviolet (UV), optical to infrared (IR) photometry of NGC 628, combined with the evolutionary stellar population synthesis, is used to derive the spatially resolved age, metallicity and reddening maps. These parameter distributions show that the bulge of this galaxy is a disk-like pseudobulge, which has the S{\'e}rsic index close to the exponential law, rich gas, and a young circumnuclear ring structure. We also discover the disk has two distinct regions with different radial age and metallicity gradients. The inner region is older and has a much steeper age gradient than the outer region of the disk. Both these two regions and the central young structure can be seen in the radial profile of the optical color. Based on the age and reddening distributions, we consider that the pseudobulge and disk are likely to have grown via the secular evolution, which is the redistribution of mass and energy through the angular momentum transport caused by the non-axisymmetric potential of the spirals. However, possible gas accretion events could affect the outer region of the disk, due to abundant H{\sc i} gas accumulating in the outer disk.Comment: 9 figures, accepted for publication in A

Scanning Tunneling Microscopy and Tunneling Luminescence of the Surface of GaN Films Grown by Vapor Phase Epitaxy

We report scanning tunneling microscopy (STM) images of surfaces of GaN films and the observation of luminescence from those films induced by highly spatially localized injection of electrons or holes using STM. This combination of scanning tunneling luminescence (STL) with STM for GaN surfaces and the ability to observe both morphology and luminescence in GaN is the first step to investigate possible correlations between surface morphology and optical properties.Comment: 12 pages, Revtex 3.0, submitted to Appl. Phys. Lett., three figures available from Jian Ma at [email protected]

Effect of hydrogen bonding and complexation with metal ions on the fluorescence of luotonin A

Fluorescence characteristics of a biologically active natural alkaloid, luotonin A (LuA), were studied by steady-state and time-resolved spectroscopic methods. The rate constant of the radiationless deactivation from the singlet-excited state diminished by more than one order of magnitude when the solvent polarity was changed from toluene to water. Dual emission was found in polyfluorinated alcohols of large hydrogen bond donating ability due to photoinitiated proton displacement along the hydrogen bond. In CH 2Cl2, LuA produced both 1:1 and 1:2 hydrogen-bonded complexes with hexafluoro-2-propanol (HFIP) in the ground state. Photoexcitation of the 1:2 complex led to protonated LuA, whose fluorescence appeared at a long wavelength. LuA served as a bidentate ligand forming 1:1 complexes with metal ions in acetonitrile. The stability of the complexes diminished in the series of Cd2+ > Zn2+ > Ag+, and upon competitive binding of water to the metal cations. The effect of chelate formation on the fluorescent properties was revealed. © 2013 The Royal Society of Chemistry and Owner Societies