Biology Reference
In-Depth Information
Oligomerization
A
H
1
R-mTq2
H
1
R-sYFP2
B
1.0
1.0
2.0
2.0
3.0
3.0
C
1.0
1.0
2.0
2.0
3.0
3.0
5 0 5
Pixel nr
5 0 5
Pixel nr
D
40
0
0
20
40
60
Time (s)
FIGURE 11.2
Working scheme for the line-FCCS data acquisition. Averaged confocal image (A) at the
equatorial plane of a HeLa cell cotransfected with H
1
R-mTurquoise2 and H
1
R-sYFP2 shows
membrane-localized fluorescence (scale bar, 5
m). A line of 16 pixels (dotted line)
perpendicular to the plasma membrane was selected for data acquisition. The intensity peaks
in the time-pixel kymographs (B), corresponding to the plasma membrane, were aligned
to correct for membrane movement during the measurement (C). For each line, the
intensities in a box of 2 pixels around the center were summed. The resulting intensity traces
(D) were auto- and cross-correlated.
m
