TBE virus was preincubated at pH 8.0 (A), 10.0 (B), and 5.4 (C), fixed with formalin, and negatively stained by phosphotungstic acid adjusted to pH 8.0 (samples A and B) or pH 6.0 (sample C). Arrows in (B) point to the rough surface generated by alkaline pH and in (C) to the bulky spikes generated by low pH treatment. All micrographs have been recorded at the same magnification. In (B) and (C), the virions lost their shell-like icosahedral envelope structure, at least at the particle surface, and as a consequence display irregular shapes that give the impression that the virus diameter is smaller than in (A). However, in all cases, the core diameter of the best-preserved virions has a similar value. In (C), the virions are aggregated, a characteristic of TBE virus maintained at low pH.