期刊论文详细信息
Viruses
Overlap Intensity: An ImageJ Macro for Analyzing the HIV-1 In Situ Uncoating Assay
Zachary Ingram1  Courtney Weatherford1  Amy E. Hulme1  Hannah Matheney1  Emma Wise1 
[1] Department of Biomedical Sciences, Missouri State University, Springfield, MO 65987, USA;
关键词: HIV-1;    capsid;    uncoating;    ImageJ;    image quantification;    fluorescent microscopy;   
DOI  :  10.3390/v13081604
来源: DOAJ
【 摘 要 】

Capsid uncoating is at the crossroads of early steps in HIV-1 replication. In recent years, the development of novel assays has expanded how HIV-1 uncoating can be studied. In the in situ uncoating assay, dual fluorescently labelled virus allows for the identification of fused viral cores. Antibody staining then detects the amount of capsid associated with each viral core at different times post-infection. Following fixed cell imaging, manual counting can be used to assess the fusion state and capsid signal for each viral core, but this method can introduce bias with increased time of analysis. To address these limitations, we developed the Overlap Intensity macro in ImageJ. This macro automates the detection of viral cores and quantification of overlapping fusion and capsid signals. We demonstrated the high accuracy of the macro by comparing core detection to manual methods. Analysis of an in situ uncoating assay further verified the macro by detecting progressive uncoating as expected. Therefore, this macro improves the accessibility of the in situ uncoating assay by replacing time-consuming manual methods or the need for expensive data analysis software. Beyond the described assay, the Overlap Intensity macro includes adjustable settings for use in other methods requiring quantification of overlapping fluorescent signals.

【 授权许可】

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