Before Ag-induced clonal expansion, nonengaging γδ TCR on nonstimulated T cells appeared to be distributed differently from their αβ TCR counterparts. Figures show topographic (left), fluorescence (middle), and merged topographic-fluorescence (right) NSOM images of αβ TCR on Vβ5⁺T cells (A) and γδ TCR on Vδ2⁺T cells (B), respectively. Note the Vβ5⁺T cell displaying the immunofluorescence TCR was derived from one of the 2 scanned Vβ5 cells as shown in the 2 lower-magnification graphs (25 × 25 μm²) inserted in the lower left (topographic) and lower right (fluorescence) bottom, respectively. See Figure 5B for NSOM images of a fluorescent Vγ2⁺ T cell and Figure S1 for more Vβ5⁺ and Vδ2⁺ T cells, and Vβ3.1⁺, and Vβ17⁺ αβ T cells. Scan size: (A) 8 × 8 μm²; (B) 7 × 7 μm². Resolution: (A,B) 500 × 500 pixel². Integration time: (A) 30 ms; (B) 10 ms. (C) The histograms showing a frequency difference in immunofluorescence TCR (size distribution) between the αβ T cell (left) and γδ T cell (right) using the Image-Plus software-based analysis. The data are the means calculated from up to 5 Vβ5⁺ cells (n = 1314) and 5 Vδ2⁺ cells (n = 5153). γδ T cells had more approximately 50-nm TCR dots distributed on cell surface than αβ T cells (P < .05).