Biomedical Engineering Reference
In-Depth Information
(a)
TPF
TPF
SHG
SHG
(b)
700
80
SHG
60
40
20
0
-20
650
Membrane potential
600
-40
-60
-80
550
1
2
3
4 5 6
Frame number
7
8
9
10
(c)
Intracellular loading
Extracellular loading
15
15
10
10
5
5
0
0
-5
-5
-10
-10
-15
-15
-125 -100 -75-50 -25 0
Membrane potential change (mV)
25
50
75 100 125
-125 -100-75 -50 -25 0
Membrane potential change (mV)
25
50
75 100 125
FIgurE 10.2 ( See color insert .) Characterization of SHG responses to membrane potential. (a) Comparison of
two-photon fluorescence (TPF) and SHG signals. TPF and SHG signals were collected simultaneously from a neu-
ron intracellularly stained with FM4-64 through the patch pipette. While TPF signals were obtained from FM4-64
dyes distributed both at the plasma membrane and cytoplasm, SHG signals were only collected from the plasma
membrane. (b) SHG signal responses to membrane potential changes. SHG signals were collected from soma of the
neuron stained with FM4-64 held at different membrane potentials. Average SHG signal intensity (red) follows the
membrane potential (black). (c) SHG signal has large and linear response to membrane potential changes. Neurons
were stained with FM4-64 either intracellularly (left) or extracellularly (right) and the SHG signal changes upon
membrane potential changes were plotted. In both cases, SHG signal shows linear response to membrane potential
changes with the amplitude of ~10% per 100 mV, but with the opposite sign. The red lines show linear fit of the data.
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