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. 2018 Nov 20;10:98–113. doi: 10.1016/j.isci.2018.11.025

Figure 2.

Figure 2

FRET Imaging of Explanted Thymic Lobes Visualizes the Heterogeneous ERK Activity of Cortical Thymocytes

(A) The experimental design for analyzing the ERK activity of thymocytes in intact thymic lobes. The thymic lobes isolated from Lck-EKAREV-NLS were examined by TPEM.

(B) Representative images of the thymic cortex. (Left) Fluorescence image of thymocytes through a BA 520-560 nm emission filter. (Right) Fluorescence image through a BA 615-675 nm emission filter showing non-T cells, including TECs. The excitation wavelength was 840 nm. Scale bar, 40 μm.

(C) Representative FRET/CFP ratio image of WT thymocytes (left) and OT-I tg thymocytes (right) in the cortex shown in intensity-modulated display (IMD) mode. Scale bar, 40 μm.

(D) The relative frequency of the FRET/CFP ratio of WT thymocytes (blue; n = 829 cells) and OT-I tg thymocytes (red; n = 721 cells). The age of mice in weeks are indicated. Similar experiments were performed with two independent mice and shown in Figure S2.