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. Author manuscript; available in PMC: 2020 Jan 1.
Published in final edited form as: J Immunol Methods. 2018 Oct 19;464:119–125. doi: 10.1016/j.jim.2018.10.010

Figure 2.

Figure 2.

Live lymph tissue slices can be stained and cultured without impacting viability and function. (a) Overview of procedure for live immunostaining. During the culture period, stimuli can be added to slices. (b) Slices showed no difference in viability after immunostaining. Calcein fluorescent intensity was measured over whole slice at 0 and 6 hours after immunostaining with anti-B220. One-way ANOVA with Bonferroni’s multiple comparisons, data representative of 2 experiments, N=3 slices per condition. (c) Immunostaining neither inhibited nor promoted an immune response in slices. Slices dual-stained for B220 and Lyve-1 were incubated for 20 hours with or without activation by PHA or anti-CD3ε. IFN-γ was quantified in the supernatant by ELISA. LOD was 93 pg/mL (dotted line). Data compiled from 2 replicate experiments, N = 10 slices per condition. Error bars denote standard deviation.