Endy:Victor3 plate reader/Measurement variation

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I measured the variation in repeated measurements on the plate reader.

Fluorescence

I measured the fluorescence of a well containing E. coli expressing YFP and a well containing untransformed MG1655. Ten measurements were taken over a 5min period.

Variation in repeated YFP measurements
Reading MG1655.YFP MG1655
1 108445 8142
2 109492 8320
3 110628 8194
4 109728 8188
5 109334 8304
6 109988 8329
7 110382 8480
8 109248 8205
9 107576 8238
10 107560 8228
Mean 109238 8263
Stdev 1016.07 93.1
CV 0.0093 0.011

Absorbance

I measured the absorbance of the same wells. Measurements were taken over the same 5min period.

Variation in repeated absorbance measurements
Reading MG1655.YFP MG1655
1 0.476 0.650
2 0.471 0.651
3 0.469 0.648
4 0.463 0.644
5 0.462 0.643
6 0.465 0.643
7 0.460 0.641
8 0.457 0.639
9 0.458 0.639
10 0.456 0.639
Mean 0.46 0.64
Stdev 0.00623 0.0042
CV 0.013 0.0065

Notes

The absorbance readings for both wells appear to decrease over time. Over a 5 min time course this is unlikely to be due to evaporation. It may be due to settling of the cells in the well. The same effect was not observed for the fluorescence readings.

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