Calculation and Comparison of the Area of Immunofluorescence of Spleen Cells in Tumor-Bearing Mice

by Bradley Miller

 

Submitted : Spring 2018

The Objective of this project is to determine how to calculate the area of fluorescence based off a fluorescent image taken in the lab. During this project, many skills were learned, including program knowledge such as excel, ImageJ, and Matlab. Images taken by an immunofluorescence microscope were analyzed on the computer using ImageJ. Through this software, a plot of mean gray value versus pixel row was created. The X and Y values of the plot were copied and pasted to an excel sheet where the Y values were shifted to have the Y-axis represent the height of fluorescence in that column of pixels of the image. A trend line of this new graph was created to acquire an equation to represent the data. This equation was then integrated from a to b values and a value for the area of fluorescence was found in the units of pixels squared. This value was then compared to the area calculated by other measuring tools in the ImageJ software.  

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