Please excuse me if, I am wrong. We quantify cytokines using ELISA or RNA analysis and western blots. It may not reliable on immunofluorescence due to lower levels of IL-6& TNFa inside cells and these two are secretory proteins.

We quantify the number of neurons expressing IL-10 bi immunofluorescence: Wrona D, Listowska M, Kubera M, Majkutewicz I, Glac W, Wojtyła-Kuchta B, Plucińska K, Grembecka B, Podlacha M. Chronic antidepressant desipramine treatment increases open field-induced brain expression and spleen production of interleukin 10 in rats. Brain Res Bull. 2013, 99:117-31. Recently we quantify also the number of cells expressing the IL-6 (not published yet, but in the attachment there is a photo of IL-6 expressing cells in the rat brain slice - orange circles and the green spots are the c-Fos expressing cells). The method of counting is described in the publication. If you have some questions please ask.

Thanks a lot for your response Chandra Dasari. But, what if we induce accumulation of these IL-6/TNFa inside the cells and then quantify?

Many Thanks for your response Dr. Irena Majkutewicz. also thanks for sharing the image of neuronal cells. DO you have any information about quantifying in in vitro cultured cells?

I think the quantification using real-time PCR is good

Unfortunatelly, I have not labelled cultured cells by immunofluorescence yet. I have experience only with tissue slices.