Please take look at below links:

http://www.amrepflow.org.au/education/links-to-omips

Also, look into Cytometry Part B Comm Clin Cytometry issues

How many colors can you handle? Are you looking for surface markers in circulating cells or will you be stimulating the cells?

There are quite a few good protocols put there, but you need to identify your target first

Here's a good 8 color panel for APCs:

Multiplexed immunophenotyping of human antigen-presenting cells in whole blood by polychromatic flow cytometry

in Nat Protoc. 2010

Many of the surface markers can also be used for other cell types is you change the lineage markers.

And more generally, here's a list of well established Optimised Multicolor Immunofluorescence Panels:

http://goo.gl/ZRsvOg

Human or mouse?

For HU following set is OK (but you need LSR II/Fortessa or other 12+ color machine)

2-3-4-8-45-19-16/56-161- dead dye (fixable, if Ki67 is used)-38-HLA-DR-Ki67 (ICS, optional)

Something like this

Gulderen and Christopher: thank you for the link to this great resource!

Gulderen: I'll definitely look into Cytometry Part B, thanks!

Christopher: I have a Navios (Beckman) so I can do 10 colours.

I would like to monitor monocytes and neutrophils, so I was thinking of the following markers: CD14, CD15, CD11b, CD16, CD62L, HLA-DR and CD142.

Don't forget Lineage-minus cocktail (to exclude T/B and other subsets out of interest which could also express HLA-DR f.e.) and dead cell dye.

10 colors are OK (1 channel for dead, 1 for Lin- and 7 for markers)

With lin- cocktail, do you mean use lineage marker antibodies all in the same fluorophor to create a "dump" channel?

Yes

Hi Ina,

not sure if this covers your request. I would recommend the following open source paper for reading

Standardization of whole blood immune phenotype monitoring for clinical trials: panels and methods from the ONE study.

Streitz M, Miloud T, Kapinsky M, Reed MR, Magari R, Geissler EK, Hutchinson JA, Vogt K, Schlickeiser S, Kverneland AH, Meisel C, Volk HD, Sawitzki B.

Transplant Res. 2013 Oct 25;2(1):17. doi: 10.1186/2047-1440-2-17.

The panels presented therein assess a rather broad view of an individual's immune system, not focussed on activation. However, the approach is possibly the most standardized immune monitoring flow assay and may contain elements or marker combinations that may be of interest for you.

Michael Kapinsky

Product Manager at Beckman Coulter

Ina,

What laser lines do you have on your navios?