scholarly journals Fluocell for Ratiometric and High-Throughput Live-Cell Image Visualization and Quantitation

2019 ◽  
Vol 7 ◽  
Author(s):  
Qin Qin ◽  
Shannon Laub ◽  
Yiwen Shi ◽  
Mingxing Ouyang ◽  
Qin Peng ◽  
...  
Keyword(s):  
High Throughput ◽  
Live Cell ◽  
Image Visualization ◽  
Cell Image ◽  
Live Cell Image

Live cell, image-based high-throughput screen to quantitate p53 stabilization and viability in human papillomavirus positive cancer cells

Virology ◽  
2021 ◽  
Author(s):  
Gustavo Martínez-Noël ◽  
Valdimara Corrêa Vieira ◽  
Patricia Szajner ◽  
Erin M. Lilienthal ◽  
Rebecca E. Kramer ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Cancer Cells ◽  
High Throughput ◽  
Live Cell ◽  
High Throughput Screen ◽  
Cell Image ◽  
Live Cell Image

Live cell image segmentation

1995 ◽  
Vol 42 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Kenong Wu ◽  
D. Gauthier ◽  
M.D. Levine
Keyword(s):  
Image Segmentation ◽  
Live Cell ◽  
Cell Image ◽  
Live Cell Image

When a single lineage is not enough: Uncertainty-Aware Tracking for spatio-temporal live-cell image analysis

2018 ◽  
Vol 35 (7) ◽  
pp. 1221-1228 ◽  
Author(s):  
Axel Theorell ◽  
Johannes Seiffarth ◽  
Alexander Grünberger ◽  
Katharina Nöh
Keyword(s):  
Image Analysis ◽  
Live Cell ◽  
Cell Image ◽  
Cell Image Analysis ◽  
Spatio Temporal ◽  
Live Cell Image

scholarly journals A high-throughput screening assay based on automated microscopy for monitoring antibiotic susceptibility of Mycobacterium tuberculosis phenotypes

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging System ◽  
Aggregate Size ◽  
Live Cell ◽  
Screening Assay ◽  
High Throughput Screening Assay

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.

scholarly journals Droplet-microarray on superhydrophobic–superhydrophilic patterns for high-throughput live cell screenings

RSC Advances ◽  
2016 ◽  
Vol 6 (44) ◽  
pp. 38263-38276 ◽  
Author(s):  
Anna A. Popova ◽  
Konstantin Demir ◽  
Titus Genisius Hartanto ◽  
Eric Schmitt ◽  
Pavel A. Levkin
Keyword(s):  
High Throughput ◽  
Microarray Platform ◽  
Live Cell ◽  
Live Cells

Droplet-microarray platform based on superhydrophobic–superhydrophilic patterning allows for miniaturized high throughput drug and transfection screenings of live cells in separated nanoliter droplets.

Identification of microRNAs that stabilize p53 in HPV-positive cancer cells

2021 ◽  
Author(s):  
Gustavo Martínez-Noël ◽  
Patricia Szajner ◽  
Rebecca E. Kramer ◽  
Kathleen A. Boyland ◽  
Asma Sheikh ◽  
...  
Keyword(s):  
Cancer Cells ◽  
High Throughput ◽  
Ubiquitin Ligase ◽  
Hela Cells ◽  
Fluorescent Protein ◽  
Human Papillomaviruses ◽  
Live Cell ◽  
Viral Oncoprotein ◽  
Protein Levels ◽  
P53 Stability

Etiologically, 5% of all cancers worldwide are caused by the high-risk human papillomaviruses (hrHPVs). These viruses encode two oncoproteins (E6 and E7) whose expression is required for cancer initiation and maintenance. Among their cellular targets are the p53 and the retinoblastoma tumor suppressor proteins. Inhibition of the hrHPV E6-mediated ubiquitylation of p53 through the E6AP ubiquitin ligase results in the stabilization of p53, leading to cellular apoptosis. We utilized a live cell high throughput screen to determine whether exogenous microRNA (miRNA) transfection had the ability to stabilize p53 in hrHPV-positive cervical cancer cells expressing a p53-fluorescent protein as an in vivo reporter of p53 stability. Among the miRNAs whose transfection resulted in the greatest p53 stabilization was 375-3p that has previously been reported to stabilize p53 in HeLa cells, providing validation of the screen. The top 32 miRNAs in addition to 375-3p were further assessed using a second cell-based p53 stability reporter system as well as in non-reporter HeLa cells to examine their effects on endogenous p53 protein levels, resulting in the identification of 23 miRNAs whose transfection increased p53 levels in HeLa cells. While a few miRNAs that stabilized p53 led to decreases in E6AP protein levels, all targeted HPV oncoprotein expression. We further examined subsets of these miRNAs for their abilities to induce apoptosis and determined whether it was p53-mediated. The introduction of specific miRNAs revealed surprisingly heterogeneous responses in different cell lines. Nonetheless, some of the miRNAs described here have potential as therapeutics for treating HPV-positive cancers. Importance Human papillomaviruses cause approximately 5% of all cancers worldwide and encode genes that contribute to both the initiation and maintenance of these cancers. The viral oncoprotein E6 is expressed in all HPV-positive cancers and functions by targeting the degradation of p53 through the engagement of the cellular ubiquitin ligase E6AP. Inhibiting the degradation of p53 leads to apoptosis in HPV-positive cancer cells. Using a high throughput live cell assay we identified several miRNAs whose transfection stabilize p53 in HPV-positive cells. These miRNAs have the potential to be used in the treatment of HPV-positive cancers.

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