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Error bars commonly appear in figures error bars encompass the lowest and high-
in publications, but experimental est values. SD is calculated by the formula
biologists are often unsure how they
should be used and interpreted. In this SD = ∑ ( X − M )2
article we illustrate some basic features n −1
of error bars and explain how they can
where X refers to the individual data
help communicate data and assist
THE JOURNAL OF CELL BIOLOGY
volume of cells from the same stock culture For n to be greater than 1, the experiment
into adjacent wells of a tissue culture plate, would have to be performed using separate
and subsequently treating them identically. stock cultures, or separate cell clones of
Although it would be possible to assay the the same type. Again, consider the popula-
plate and determine the means and errors of tion you wish to make inferences about—it
the replicate wells, the errors would reflect is unlikely to be just a single stock culture.
the accuracy of pipetting, not the reproduc- Whenever you see a figure with very small
iblity of the differences between the experi- error bars (such as Fig. 3), you should ask
mental cells and the control cells. For yourself whether the very small variation
replicates, n = 1, and it is therefore in- implied by the error bars is due to analysis
appropriate to show error bars or statistics. of replicates rather than independent sam-
If an experiment involves triplicate ples. If so, the bars are useless for making
cultures, and is repeated four independent the inference you are considering.
times, then n = 4, not 3 or 12. The varia- Sometimes a figure shows only the
tion within each set of triplicates is related data for a representative experiment, imply-
to the fidelity with which the replicates ing that several other similar experiments
were created, and is irrelevant to the hy- were also conducted. If a representative ex-
Figure 4. Inferential error bars. Means with pothesis being tested. periment is shown, then n = 1, and no error
SE and 95% CI error bars for three cases, rang- To identify the appropriate value for bars or P values should be shown. Instead,
ing in size from n = 3 to n = 30, with descrip- n, think of what entire population is being the means and errors of all the independent
tive SD bars shown for comparison. The small
black dots are data points, and the large dots sampled, or what the entire set of experi- experiments should be given, where n is the
double the SE bars just touch, P is close to the SE or 95% CI calculated from those
0.05 (Fig. 5, leftmost panel). If n is 10 or differences. If that 95% CI does not in-