Sorry, I can´t see the video. But if you only need to trak a few objects and get their speed (avg or in one timepoint) and direcction, you can try the Manual Tracking Plugging. It is very easy to use it and it has a nice manual online.

http://fiji.sc/Manual_Tracking

Hope this helps.

Thank you very much.

The problem is that what i am tracking is not really like a particle.

Please can you take a look at the video?

Unfortunately, i am finding it difficult to upload the video.

Hi Uchenna, even if your tracking target is not a particle, you can still use the manual tracking Victor suggested. You will need to select the target frame by frame manually, and the plugin will "lock" on the target depending on your setting. Also, you can set the scaling factor so that the resulting data table gives you pixel distance converted to calibrated distance per time between frames - which is your velocity... Good luck.

If you are okay with using matlab, there is qFSM package that is really suitable for actin flow tracking (even without speckle-ness), which is developed in Danuser lab:

http://lccb.hms.harvard.edu/software.html

The package has both flow-tracking (that uses PIV-type flow estimation) and particle-tracking mode that allows quantification of actin polymerization/depolymerization. But for your purpose, flow-tracking is good enough.

Thank you very much Sangyoon J Han and Clemens Alt. I will try as you guys suggested! Thanks again!!