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Colony-forming unit (CFU) assay

MH M. T. Heemskerk
CK C. J. Korbee
JE J. J. Esselink
CS C. Carvalho dos Santos
SV S. van Veen
IG I. F. Gordijn
FV F. Vrieling
KW K. V. Walburg
CE C. G. Engele
KD K. Dijkman
LW L. Wilson
FV F. A. W. Verreck
TO T. H. M. Ottenhoff
MH M. C. Haks
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Cells were lysed in H2O containing 0.05% SDS (ThermoFisher Scientific). Lysates of Mtb-infected cells were serially diluted in steps of fivefold in 7H9 broth and 10 μl droplets were spotted onto square Middlebrook 7H10 agar plates. Plates were incubated at 37 °C for 12–14 days and bacterial colonies quantified using a microscope with a magnification of 2.5 times to enhance early detection of bacterial growth. Lysates of Stm-infected cells were serially diluted in LB broth, thereafter 10 μl droplets were spotted onto square LB agar plates and incubated overnight at 37 °C.

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