Java

Manual tracking using Trackmate plugin (comes with FIji, so no installation required if you are using Fiji). 

This method was originally designed to track objects (not necessarily spots) already identified in 2D 
frames and has been applied previously to particle tracking and analysis in high-speed atomic force microscopy image series.

JAMA is a basic linear algebra package for Java. It provides user-level classes for constructing and manipulating real, dense matrices. It is meant to provide sufficient functionality for routine problems, packaged in a way that is natural and understandable to non-experts. It is intended to serve as the standard matrix class for Java, and will be proposed as such to the Java Grande Forum and then to Sun. A straightforward public-domain reference implementation has been developed by the MathWorks and NIST as a strawman for such a class. We are releasing this version in order to obtain public comment. There is no guarantee that future versions of JAMA will be compatible with this one.

The phase contrast microscopy segmentation toolbox (PHANTAST) is a collection of open-source algorithms and tools for the processing of phase contrast microscopy (PCM) images. It was developed at University College London's department of Biochemical Engineering and CoMPLEX.

An ImageJ plugin for DEFCoN, the fluorescence spot counter based on fully convolutional neural networks