Programs for cell counting in single cell microscopy

[roodkontjeaapje]

Hi everyone,

 

I'm not sure if this is the right forum for this type of question (please give me your recommendations if I'm wrong), but here we go.

I work in a lab and we sometimes perform this single cell microscopy assay, where yeast cells are spread on a micro-array after being incubated with both a potentially fungicidal agent and a substance to mark dead cells. The micro-array with the cells on it is visualized through a microscope while a laser shines on it. Like this, dead cells will light up thanks to a fluorescent marker agent (called PI if I'm not mistaking) and the magnified image of the entire micro-array is displayed on a computer screen. Photos of the micro-array are taken regularly (e.g. every 15 minutes) and you can see how many additional yeast cells are killed over time simply by counting them on every picture. And that's a hell of a lot of work: you have to count all the dead cells (which can be identified since they light up) on the entire array and do this for multiple pictures that are taken sequentially.

 

My question, is there a computer program that can do this counting for me? This program should pretty much be able to count shiny dots on a picture, which are dead cells. But a few conditions have to be met:

-The micro-array is covered with microscopic wells (which all have the same depth and are regularly interspaced, so they're all the same distance from one another) and only the cells -inside- those wells should be counted;

-Some debris particles might light up as well, these should not be counted

-Only cells that light up with a certain minimal intensity are dead and only those should be counted.

-The edges of the wells tend to light up as well and the program should be able to distinguish this from cells (well are just empty circles that light up and cells are more like solidly colored).

Can anyone think of a program like this, or help me find someone who can design it?

 

Many thanks.