Not sure if this is the correct place to start this topic, but I need some help. I'm doing my research on the induction of macrophage in the peritoneal cavity of mice. I'm doing IP injections on 3 groups of mice. One group will be injected with thioglycollate alone. Group 2 is injected with thioglycollate along with a catalase enzyme and Group 3 is going to be injected with thioglycollate along with a Cholesterol oxidase enzyme. Basically, I'm just going to do cell counts and staining to see which group has the most induction.
I know how to do the injections and I know how to retrieve the cells. My questions are this:
Once I have collected the cells, washed them, and spinned them down, do I need to vortex them to re-suspend the cells to take a sample for slides?
I'm using Wright's stain for staining.
When doing my cell counts with a hemocytometer, do the cells have to be stained first? Or will I be able to distinguish and identify the macrophages? I know the cells will be from the suspension.
Also when doing the cell counts, do I have to do dilutions, or can I just pipette straight from the sample.
I guess these sorta sound like silly questions that I should know the answers to, but I've grown a little rusty on it.