Use of a Double-Barreled Biolistic Device and Counting Software for Comparing Performance Between Plant Transfection Procedures
Date
2020-12
Authors
Kirscht, Tyler
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Abstract
The accuracy of plant cell characterization is improved through the optimization of CellProfiler software, where images of fluorescent gene expression are analyzed to determine the efficacy of a gene delivery system. CellProfiler is an open-source cell characterization software made up of various modules that process images. These modules require their settings to be fine-tuned in order to accurately identify, separate, and count the cells with modified DNA. These inputs affect the various steps which the program uses to count the cells. The brightness threshold is the first, in which the program runs an algorithmic thresholding method to determine which cells are significant. Several of these thresholding methods were tested, along with a threshold correction factor that empirically adjusts the calculated threshold value. An enhancement filter and Gaussian blur were also implemented to suppress background noise while making the cell features more prominent. Additionally, the cell size parameter was adjusted for effective brightness. Finally, the program uses de-clumping to separate and identify cells that are grouped together, based on several empirically determined factors. Overall, the accuracy of the software was significantly improved, and the lessons learned here can be taken into future plant cell characterization projects.