Hi all, I am studying structures that are similar to focal adhesions called 
podosomes. They are rich in actin and make an actin core of about 1 micron in 
diameter.
 
These podosomes have a lifetime about about 2 mins (that is, it takes 2 mins 
for a "dot" in the cell to become visible and dissappear again).  

At this stage I use a CSU-X on an Andor system (63x, binned 2x2 Ocra ER) 
but I find it very hard to resolve because there is an "actin cloud" that is 
found within the podosome belt (which you see surrounds the gigantic 
osteoclast cell). I will be trying TIRF soon. 

I also do not have any software for analyzing this (apart from going frame by 
frame in image j).  

As such, would you have any suggestions as to the best way of acquiring the 
images (CSU, point scanner etc) and software for this work? 

I would also be interested in communicating with anyone who does this sort of 
particle tracking. 

Regards Sebastien