Fig. 2 Linear relationship between the number of Leptospira and resazurin reduction. Exponentially growing
Leptospira were serially diluted to obtain the indicated concentration and 100 μl of Leptospira were
transferred in a flat bottom 96-well plates (TPP
® Tissue Culture Test Plate in (a) and (b); μClear Black
96-well plate in (c)). 80 μl of EMJH medium and 20 μl of 10Â resazurin solution (provided by the Alamar Blue
®
Assay) were added. The plates were incubated 24 h at 30
C. (a) The change of the blue resazurin solution into
a pink resorufin solution upon reduction is shown. (b) The percentage of resazurin reduction was calculated as
described in Fig. 3 and plotted in function of the bacterial concentration (step 7 in Subheading 3.3). (c) The
fluorescence was measured (λ exc ¼ 560 nm, λ em ¼ 590 nm) and plotted in function of the bacterial
concentration (step 7 in Subheading 3.3)
218
Cle ´ mence Mouville and Nadia Benaroudj
Leptospira were serially diluted to obtain the indicated concentration and 100 μl of Leptospira were
transferred in a flat bottom 96-well plates (TPP
® Tissue Culture Test Plate in (a) and (b); μClear Black
96-well plate in (c)). 80 μl of EMJH medium and 20 μl of 10Â resazurin solution (provided by the Alamar Blue
®
Assay) were added. The plates were incubated 24 h at 30
C. (a) The change of the blue resazurin solution into
a pink resorufin solution upon reduction is shown. (b) The percentage of resazurin reduction was calculated as
described in Fig. 3 and plotted in function of the bacterial concentration (step 7 in Subheading 3.3). (c) The
fluorescence was measured (λ exc ¼ 560 nm, λ em ¼ 590 nm) and plotted in function of the bacterial
concentration (step 7 in Subheading 3.3)
218
Cle ´ mence Mouville and Nadia Benaroudj