Abstract
Counting labeled cells, after immunofluorescence or expression of a genetically fluorescent reporter protein, is frequently used to quantify viral infection. However, this can be very tedious without a high content screening apparatus. For this reason, we have developed QuantIF, an ImageJ macro that automatically determines the total number of cells and the number of labeled cells from two images of the same field, using DAPI- and specific-stainings, respectively. QuantIF can automatically analyze hundreds of images, taking approximately one second for each field. It is freely available as supplementary data online at MDPI.com and has been developed using ImageJ, a free image processing program that can run on any computer with a Java virtual machine, which is distributed for Windows, Mac, and Linux. It is routinely used in our labs to quantify viral infections in vitro, but can easily be used for other applications that require quantification of labeled cells.
Highlights
When evaluating viral infections in vitro, fluorescence microscopy is commonly used to monitor the expression of a viral protein following immunostaining
We present QuantIF, an ImageJ macro for determining the percentage of fluorescent cells following immunofluorescence staining
QuantIF is a free, simple, and robust automated tool to estimate the proportion of virally infected cells after immunofluorescence
Summary
When evaluating viral infections in vitro, fluorescence microscopy is commonly used to monitor the expression of a viral protein following immunostaining. Rasband at the National Institute of Health, and has become a valuable tool for researchers [1,2] It is a Java-based software that can run on any computer using a Java virtual machine. A major strength of ImageJ is the possibility to record macros that enable the automatization of image analysis. In this technical note, we present QuantIF, an ImageJ macro for determining the percentage of fluorescent cells following immunofluorescence staining. When the macro is run, it automatically processes all images in the folder, taking around one second to analyze both images of each field. All results are saved as a “.xls” file that can be processed for statistical analysis
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
