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The use of Transmission Electron Microscopy (TEM) to characterize the microstructure of a material continues to grow in importance as technological advancements become increasingly more dependent on nanotechnology 1. Since nanoparticle properties such as size (diameter) and size distribution are often important in determining potential applications, a particle analysis is often performed on TEM images. Traditionally done manually, this has the potential to be labor intensive, time consuming, and subjective 2. To resolve these issues, automated particle analysis routines are becoming more widely accepted within the community 3. When using such programs, it is important to compare their performance, in terms of functionality and cost. The primary goal of this study was to apply one such software package, ImageJ to grayscale TEM images of nanoparticles with known size. A secondary goal was to compare this popular open-source general purpose image processing program to two commercial software packages. After a brief investigation of performance and price, ImageJ was identified as the software best suited for the particle analysis conducted in the study. While many ImageJ functions were used, the ability to break agglomerations that occur in specimen preparation into separate particles using a watershed algorithm was particularly helpful 4. © 2009 SPIE-IS&T.