[Histonet] IF staining on peritoneal macrophages

[Mauricio Avigdor]

Greetings all,

I am trying to do immunofluorescence on peritoneal macrophages. I am having
a couple of issues that I was hoping one of you could help me resolve.

Firstly, I am having uneven results with the Cytospin. Cells tend to get
washed off the slides during rinses. Does anyone have tips on how to make
cells stick a little better to Cytoslides? Secondly, I have not yet found a
satisfactory method for fixation. In order to prevent loss of cells when
dipping the slides into fixatives, I am having to air dry the slides before
I can do anything to them. I tried the Shandon Collection Fluid (ethanol,
isopropanol, carbowax) with great success, but it killed the fluorescence.

I am leaning towards spinning the lavage fluid until I get a pellet and then
resuspending in PBS with a little BSA added. I hope this makes cells stick
well enough that I can put the slides in formalin or acetone.

Any thoughts are appreciated!