Fig. 4

Germination, outgrowth, elongation, and growth of multiple individual untreated B. subtilis PS533 spores without a, c or with b, d a 10 day high vacuum treatment. Untreated or high vacuum-treated spores were cultured on an LB medium agar pad on a microscope coverslip at 37 °C and bright-field images were recorded every 60 s for 4 h as described in Methods. a, b Bright-field images of individual spores at various culture times. In a panels the red and black arrows show images of two spores that germinated and grew; in panel b the blue, red, and black arrows show the images of spores that never germinated, germinated but did not grow, and germinated and grew, respectively. c, d Cell length vs time for 7 individual untreated and high vacuum-treated spores that germinated and grew, and arrows denote the T_elong times