Optimization of the minimal sample protocol.
The panels describe
testing of the need for a carrier channel, carrier channel input size,
and position. (a) Panel shows an overview of cell numbers used for
different purposes in the protocol and comparisons. (b) Comparison
of TMT setups for using 1000 cells with and without carrier channel
and using 60 000, 20 000, and 10 000 cells for
the carrier channel. The graph shows the numbers of protein groups
detected using the experimental setup below. The error bars show standard
deviation. (c) Effects of carrier channel on neighboring channels
at 1 Thompson distance using a 10plex TMT experiment with empty channels
2–8 and a sample from 10 000 cells in channels 1 and
10. ESC—embryonic stem cell, MN—motor neuron, FACS—fluorescence-activated
cell sorting, and TMT—tandem mass tag.