Validation of the robustness of the G/R ratiometric approach
to
different hardware (cell phone cameras) and lighting conditions. (a–g)
Enlarged and cropped color images (top two rows of each individual
panel) captured by an unmodified cell phone camera from positive (+)
and negative (−) RT-LAMP reactions at 2-fold increases in EBT
concentration from 10.9 μM to 1.4 mM (1 = 0.011 mM; 2 = 0.022
mM; 3 = 0.044 mM, 4 = 0.088 mM, 5 = 0.175 mM; 6 = 0.35 mM; 7 = 0.7
mM; 8 = 1.4 mM). Positive wells are blue and negative wells are purple.
After G/R ratiometric processing (bottom two rows of each individual
panel), negative wells are black. Regions I, II, III in each panel
indicate the effect of dye concentration: (II) acceptable concentration
range for visualization (green regions); (I) concentrations too low
for visualization (white regions); and (III) concentrations too high
for visualization (red regions). (a–d) Images captured by four
common cell phones under fluorescent light: (a) Apple iPhone 4S, (b)
HTC inspire 4G, (c) Motorola Moto G, and (d) Nokia 808 PureView. (e–g)
Images captured by an Apple iPhone 4S under three additional light
conditions: (e) incandescent light, (f) direct sunlight, and (g) indirect
sunlight. All experiments were performed with HCV RNA as a clinically
relevant target. All images were acquired with unmodified cell phone
cameras. Detailed information for the G/R ratiometric process (Figure
S2) and additional cell phone camera images (Figure S3) are provided
in the Supporting Information.