scholarly journals Systematic Quantification of Cell Confluence in Human Normal Oral Fibroblasts

2020 ◽  
Vol 10 (24) ◽  
pp. 9146
Author(s):  
Ching-Hsiang Chiu ◽  
Jyh-Der Leu ◽  
Tzu-Ting Lin ◽  
Pin-Hua Su ◽  
Wan-Chun Li ◽  
...  

Background: The accurate determination of cell confluence is a critical step for generating reasonable results of designed experiments in cell biological studies. However, the cell confluence of the same culture may be diversely predicted by individual researchers. Herein, we designed a systematic quantification scheme implemented on the Matlab platform, the so-called “Confluence-Viewer” program, to assist cell biologists to better determine the cell confluence. Methods: Human normal oral fibroblasts (hOFs) seeded in 10 cm culture dishes were visualized under an inverted microscope for the acquisition of cell images. The images were subjected to the cell segmentation algorithm with top-hat transformation and the Otsu thresholding technique. A regression model was built using a quadratic model and shape-preserving piecewise cubic model. Results: The cell segmentation algorithm generated a regression curve that was highly correlated with the cell confluence determined by experienced researchers. However, the correlation was low when compared to the cell confluence determined by novice students. Interestingly, the cell confluence determined by experienced researchers became more diverse when they checked the same images without a time limitation (up to 1 min). Conclusion: This tool could prevent unnecessary human-made mistakes and meaningless repeats for novice researchers working on cell-based studies in health care or cancer research.

Author(s):  
Kris Gillis ◽  
Jean-Yves Wielandts ◽  
Gabriela Hilfiker ◽  
Louisa O'Neill ◽  
Alina Vlase ◽  
...  

Introduction. During left bundle branch area pacing (LBBAP) lead implantation, intermittent monitoring of unipolar pacing characteristics validates LBB capture and can detect septal perforation. We aimed to demonstrate that continuous uninterrupted unipolar pacing from an inserted lead stylet (LS) is feasible and facilitates LBBAP implantation. Methods. Thirty patients (mean age 76 ± 14 years) were implanted with stylet-driven pacing lead (Biotronik Solia S60). In 10 patients (validation-group) conventional, interrupted implantation was performed, with comparison of unipolar pacing characteristics between LS and connector-pin (CP)-pacing after each rotation step. In 20 patients (feasibility-group) performance and safety of uninterrupted implantation during continuous pacing from the LS were analyzed. Results. In the validation-group, LS and CP-pacing impedances were highly correlated (R=0.95, p<0.0001, bias 12±37Ω). Pacing characteristics from LS and CP showed comparable sensed electrograms and paced QRS morphologies. In the feasibility-group, continuous LS-pacing allowed beat-to-beat monitoring of impedance and QRS morphology to guide implantation. This resulted in successful LBBAP in all patients, after a mean of 1±0 attempts, with mean threshold 0.81 ± 0.4V, median sensing 6.5mV [IQR 4.4-9.5] and mean impedance 624 ± 101Ω, and positive LBBAP-criteria with median paced QRS duration 120ms [IQR 112-152ms] and median pLVAT 73ms [IQR 68-80.5ms]. No septal perforation occurred. Conclusion. Unipolar pacing from the LS allows accurate determination of pacing impedance and generates similar paced QRS morphologies and equal sensed electrograms, compared to CP pacing. Continuous LS pacing allows real-time monitoring of impedance and paced QRS morphology, which facilitates a safe and successful LBBAP lead implantation.


2021 ◽  
Author(s):  
Chibuzo Onwukaeme ◽  
Won-Jin Choi ◽  
Han-Youl Ryu

Abstract We investigate the junction temperature measurements for GaN-based blue light emitting diodes (LEDs) using nonlinear dependence of the forward voltage ( V f ) on temperature. Unlike the conventional linear model of the dependence of V f on temperature, the modeling of the temperature dependent V f with a quadratic function showed good agreements with measured data in the temperature range between 20 and 100 o C. Using the proposed quadratic model, the junction temperature and thermal resistance of the measured LED could be accurately determined as the ambient temperature varied. It was observed that the junction temperature increment remained almost unchanged as the ambient temperature increased from 20 to 80 o C, which could be attributed to the interplay between the decrease in series resistance and the increase in non-radiative recombination with increasing temperature. The presented method for accurate determination of the junction temperature is expected to be advantageously employed for the thermal management of high-power LEDs.


Author(s):  
R.D. Leapman ◽  
P. Rez ◽  
D.F. Mayers

Microanalysis by EELS has been developing rapidly and though the general form of the spectrum is now understood there is a need to put the technique on a more quantitative basis (1,2). Certain aspects important for microanalysis include: (i) accurate determination of the partial cross sections, σx(α,ΔE) for core excitation when scattering lies inside collection angle a and energy range ΔE above the edge, (ii) behavior of the background intensity due to excitation of less strongly bound electrons, necessary for extrapolation beneath the signal of interest, (iii) departures from the simple hydrogenic K-edge seen in L and M losses, effecting σx and complicating microanalysis. Such problems might be approached empirically but here we describe how computation can elucidate the spectrum shape.The inelastic cross section differential with respect to energy transfer E and momentum transfer q for electrons of energy E0 and velocity v can be written as


Author(s):  
M.A. Gribelyuk ◽  
M. Rühle

A new method is suggested for the accurate determination of the incident beam direction K, crystal thickness t and the coordinates of the basic reciprocal lattice vectors V1 and V2 (Fig. 1) of the ZOLZ plans in pixels of the digitized 2-D CBED pattern. For a given structure model and some estimated values Vest and Kest of some point O in the CBED pattern a set of line scans AkBk is chosen so that all the scans are located within CBED disks.The points on line scans AkBk are conjugate to those on A0B0 since they are shifted by the reciprocal vector gk with respect to each other. As many conjugate scans are considered as CBED disks fall into the energy filtered region of the experimental pattern. Electron intensities of the transmitted beam I0 and diffracted beams Igk for all points on conjugate scans are found as a function of crystal thickness t on the basis of the full dynamical calculation.


Author(s):  
F.A. Ponce ◽  
H. Hikashi

The determination of the atomic positions from HRTEM micrographs is only possible if the optical parameters are known to a certain accuracy, and reliable through-focus series are available to match the experimental images with calculated images of possible atomic models. The main limitation in interpreting images at the atomic level is the knowledge of the optical parameters such as beam alignment, astigmatism correction and defocus value. Under ordinary conditions, the uncertainty in these values is sufficiently large to prevent the accurate determination of the atomic positions. Therefore, in order to achieve the resolution power of the microscope (under 0.2nm) it is necessary to take extraordinary measures. The use of on line computers has been proposed [e.g.: 2-5] and used with certain amount of success.We have built a system that can perform operations in the range of one frame stored and analyzed per second. A schematic diagram of the system is shown in figure 1. A JEOL 4000EX microscope equipped with an external computer interface is directly linked to a SUN-3 computer. All electrical parameters in the microscope can be changed via this interface by the use of a set of commands. The image is received from a video camera. A commercial image processor improves the signal-to-noise ratio by recursively averaging with a time constant, usually set at 0.25 sec. The computer software is based on a multi-window system and is entirely mouse-driven. All operations can be performed by clicking the mouse on the appropiate windows and buttons. This capability leads to extreme friendliness, ease of operation, and high operator speeds. Image analysis can be done in various ways. Here, we have measured the image contrast and used it to optimize certain parameters. The system is designed to have instant access to: (a) x- and y- alignment coils, (b) x- and y- astigmatism correction coils, and (c) objective lens current. The algorithm is shown in figure 2. Figure 3 shows an example taken from a thin CdTe crystal. The image contrast is displayed for changing objective lens current (defocus value). The display is calibrated in angstroms. Images are stored on the disk and are accessible by clicking the data points in the graph. Some of the frame-store images are displayed in Fig. 4.


1996 ◽  
Vol 75 (05) ◽  
pp. 772-777 ◽  
Author(s):  
Sybille Albrecht ◽  
Matthias Kotzsch ◽  
Gabriele Siegert ◽  
Thomas Luther ◽  
Heinz Großmann ◽  
...  

SummaryThe plasma tissue factor (TF) concentration was correlated to factor VII concentration (FVIIag) and factor VII activity (FVIIc) in 498 healthy volunteers ranging in age from 17 to 64 years. Immunoassays using monoclonal antibodies (mAbs) were developed for the determination of TF and FVIIag in plasma. The mAbs and the test systems were characterized. The mean value of the TF concentration was 172 ± 135 pg/ml. TF showed no age- and gender-related differences. For the total population, FVIIc, determined by a clotting test, was 110 ± 15% and the factor VIlag was 0.77 ± 0.19 μg/ml. FVII activity was significantly increased with age, whereas the concentration demonstrated no correlation to age in this population. FVII concentration is highly correlated with the activity as measured by clotting assay using rabbit thromboplastin. The ratio between FVIIc and FVIIag was not age-dependent, but demonstrated a significant difference between men and women. Between TF and FVII we could not detect a correlation.


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