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Figure 1. Time-course of the accumulation of the reaction product.
A. platensis NIES-39 filaments (291 ± 39 filaments in 50 µL of SOT medium)
were mixed with 5 µL of MTT solution (5 mg/mL in H2O) and cultured. After the
indicated periods, filaments were recovered by centrifugation, and formazan
product in the cells was dissolved in acid-isopropanol. Relative amounts of
formazan product were determined by measuring absorbance at 570 nm. Viable
(open circles) and heat-killed cells (closed circles) were used for the reactions. The
means and standard errors of the means are shown (n = 5).
Figure 2. Dynamic range and sensitivity of MTT assay.
Varied numbers of A. platensis filaments were incubated with MTT in microfuge
tubes, and accumulation of the reaction product was determined. (a) A dilution
series of A. platensis filaments were subjected to the assay. (b) Filaments were
individually picked using a micropipette, and the indicated numbers of filaments
that were suspended in 50 µL of SOT medium were subjected to the assay. The
means and standard errors of the means are shown (n = 5). Linear regression
analysis was performed using GraphPad Prism 8 (GraphPad Software, San Diego).
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Figure 3. Viability analysis of A. platensis after heat-treatment.
(a) A. platensis filaments (approximately 400 filaments) were heated for 30 min at various
temperatures in 50 µL of SOT medium and then cultured in 1 mL of the same medium.
Viability was determined by comparing the growth curves of heat-treated and control
samples. (b) MTT assay was performed soon after the heat-treatment in 50 µL of SOT
medium. Relative OD570 values relative to control samples (30°C) were calculated and
plotted by setting the mean value of the control samples (0.207 at 30°C) as 100. (c)
Samples were maintained for 16 h under standard culture conditions after heat-treatment.
Then, MTT assay was performed. OD570 values relative to the mean OD570 value of the
control samples (0.269 at 30ºC) were calculated and plotted as in (b). For each data point,
the mean and the standard error of the mean is shown (n = 5).
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