scholarly journals High-throughput quantitative detection of basal autophagy and autophagic flux using image cytometry

BioTechniques ◽  
2019 ◽  
Vol 67 (2) ◽  
pp. 70-73 ◽  
Author(s):  
Gopika SenthilKumar ◽  
Justin H Skiba ◽  
Randall J Kimple
Keyword(s):  
High Throughput ◽  
Image Cytometry ◽  
Quantitative Detection ◽  
Autophagic Flux

scholarly journals High-throughput quantitative detection of basal autophagy and autophagic flux using image cytometry

2019 ◽  
Author(s):  
Gopika SenthilKumar ◽  
Justin H. Skiba ◽  
Randall J. Kimple
Keyword(s):  
Acridine Orange ◽  
High Throughput ◽  
Late Stage ◽  
Image Cytometry ◽  
Alternative Methods ◽  
Quantitative Detection ◽  
Autophagic Flux ◽  
Objective Method ◽  
Acridine Orange Staining ◽  
Transmission Electron

AbstractQuantitative assessment of changes in macro-autophagy is often performed through manual quantification of the number of LC3B foci in immunofluorescence microscopy images. This method is highly laborious, subject to image-field selection and foci-counting bias, and is not sensitive for analyzing changes in basal autophagy. Alternative methods such as flow cytometry and transmission electron microscopy require highly specialized, expensive instruments and time-consuming sample preparation. Immunoblots are prone to exposure-related variations and noise that prevent accurate quantification. We report a high-throughput, inexpensive, reliable, and objective method for studying basal level and flux changes in late-stage autophagy using image cytometry and acridine orange staining.Methods summaryA high-throughput, inexpensive, reliable, and objective method for studying both basal autophagy and autophagic flux is reported. This approach uses acridine orange staining of late-stage autophagy and image cytometry to quantify autophagy.

scholarly journals High-throughput viral microneutralization method for feline coronavirus using image cytometry

2020 ◽  
Vol 286 ◽  
pp. 113979
Author(s):  
Morgan Pearson ◽  
Alora LaVoy ◽  
Leo Li-Ying Chan ◽  
Gregg A. Dean
Keyword(s):  
High Throughput ◽  
Image Cytometry ◽  
Feline Coronavirus

scholarly journals Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

Viruses ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 655 ◽  
Author(s):  
Yíngyún Caì ◽  
Masaharu Iwasaki ◽  
Brett Beitzel ◽  
Shuīqìng Yú ◽  
Elena Postnikova ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
High Throughput ◽  
Reverse Genetics ◽  
Fluorescent Protein ◽  
Cultured Cells ◽  
Neutralizing Antibody ◽  
Quantitative Detection ◽  
Lassa Virus ◽  
Wild Type ◽  
Green Fluorescent

Lassa virus (LASV), a mammarenavirus, infects an estimated 100,000–300,000 individuals yearly in western Africa and frequently causes lethal disease. Currently, no LASV-specific antivirals or vaccines are commercially available for prevention or treatment of Lassa fever, the disease caused by LASV. The development of medical countermeasure screening platforms is a crucial step to yield licensable products. Using reverse genetics, we generated a recombinant wild-type LASV (rLASV-WT) and a modified version thereof encoding a cleavable green fluorescent protein (GFP) as a reporter for rapid and quantitative detection of infection (rLASV-GFP). Both rLASV-WT and wild-type LASV exhibited similar growth kinetics in cultured cells, whereas growth of rLASV-GFP was slightly impaired. GFP reporter expression by rLASV-GFP remained stable over several serial passages in Vero cells. Using two well-characterized broad-spectrum antivirals known to inhibit LASV infection, favipiravir and ribavirin, we demonstrate that rLASV-GFP is a suitable screening tool for the identification of LASV infection inhibitors. Building on these findings, we established a rLASV-GFP-based high-throughput drug discovery screen and an rLASV-GFP-based antibody neutralization assay. Both platforms, now available as a standard tool at the IRF-Frederick (an international resource), will accelerate anti-LASV medical countermeasure discovery and reduce costs of antiviral screens in maximum containment laboratories.

scholarly journals One-step high-throughput assay for quantitative detection of β-galactosidase activity in intact Gram-negative bacteria, yeast, and mammalian cells

BioTechniques ◽  
2006 ◽  
Vol 40 (4) ◽  
pp. 433-440 ◽  
Author(s):  
Faustino Vidal-Aroca ◽  
Michele Giannattasio ◽  
Elisa Brunelli ◽  
Alessandro Vezzoli ◽  
Paolo Plevani ◽  
...  
Keyword(s):  
High Throughput ◽  
Mammalian Cells ◽  
Quantitative Detection ◽  
Gram Negative Bacteria ◽  
Galactosidase Activity ◽  
Gram Negative ◽  
One Step ◽  
High Throughput Assay

High-throughput two-step LNA real time PCR assay for the quantitative detection and genotyping of HPV prognostic-risk groups

2009 ◽  
Vol 45 (4) ◽  
pp. 304-310 ◽  
Author(s):  
Elisa Leo ◽  
Simona Venturoli ◽  
Monica Cricca ◽  
Monica Musiani ◽  
Marialuisa Zerbini
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Risk Groups ◽  
Quantitative Detection ◽  
Pcr Assay

High-Throughput Image Cytometry for Rare Cell Detection

2013 ◽  
Author(s):  
Keisuke Goda ◽  
Genshun Yu ◽  
Yoshifumi Wakisaka ◽  
Hirofumi Kobayashi
Keyword(s):  
High Throughput ◽  
Image Cytometry ◽  
Cell Detection

scholarly journals Subnuclear foci quantification using high-throughput 3D image cytometry

2015 ◽  
Author(s):  
Dushan N. Wadduwage ◽  
Marcus Parrish ◽  
Heejin Choi ◽  
Bevin P. Engelward ◽  
Paul Matsudaira ◽  
...  
Keyword(s):  
High Throughput ◽  
Image Cytometry ◽  
3D Image

A simple and high-throughput luciferase immunosorbent assay for both qualitative and semi-quantitative detection of anti-HIV-1 antibodies

2019 ◽  
Vol 263 ◽  
pp. 9-15 ◽  
Author(s):  
Haiying Wang ◽  
Qundi Cai ◽  
Yuanhao Liang ◽  
Jingwei Shui ◽  
Shixing Tang
Keyword(s):  
Immunosorbent Assay ◽  
High Throughput ◽  
Quantitative Detection ◽  
Anti Hiv ◽  
Hiv 1

scholarly journals Subnuclear foci quantification using high-throughput 3D image cytometry

2015 ◽  
Author(s):  
Dushan N. Wadduwage ◽  
Marcus Parrish ◽  
Heejin Choi ◽  
Bevin P. Engelward ◽  
Paul Matsudaira ◽  
...  
Keyword(s):  
High Throughput ◽  
Image Cytometry ◽  
3D Image

scholarly journals High-throughput detection of DNA double-strand breaks using image cytometry

BioTechniques ◽  
2015 ◽  
Vol 58 (1) ◽  
Author(s):  
Tyler L. Fowler ◽  
Alison M. Bailey ◽  
Bryan P. Bednarz ◽  
Randall J. Kimple
Keyword(s):  
High Throughput ◽  
Image Cytometry ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Strand Breaks ◽  
High Throughput Detection
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