Fig. 4: Hemodynamic and motion correction can be achieved with a red fluorescent protein and a single excitation light source. | Nature Communications

Fig. 4: Hemodynamic and motion correction can be achieved with a red fluorescent protein and a single excitation light source.

From: Widefield imaging of rapid pan-cortical voltage dynamics with an indicator evolved for one-photon microscopy

Fig. 4

a 5-Hz oscillations observed in JEDI-1P-Kv and RFP reference channels in lightly anesthetized mice correspond to the heart rate (shown in EKG) and reflect fast blood volume changes (fast hemodynamics). JEDI-1P-Kv and RFP optical traces are from the same 2 × 2-pixel ROI. The regression pipeline for anesthetized mice is as in panel (c) but used 4–20, 1–4, 0–1 Hz as the three bandpass filters. b Successful removal of fast hemodynamic signals in a lightly anesthetized animal. n = 25 trials from a representative mouse. The heartbeat peak at ~5 Hz (arrow) was removed in the regressed trace (blue). c Signal pre-processing workflow when recording from awake animals. The reference channel was filtered to three different frequency ranges for regression: 10–30 Hz—which includes the heartbeat frequency in the awake condition, 1–10 Hz, and 0–1 Hz. These ranges produced efficient regression in our awake mice, but different ranges might be optimal in other studies. d Step-wise regression on representative traces removed fluorescence signals unrelated to voltage. An example of pixel-wise regression is shown in Supplementary Fig. 9.

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