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Oral Dosing of Chemical Indicators for In Vivo Monitoring of Ca2+ Dynamics in Insect Muscle

Figure 3

Ca2+ dynamics and muscle displacement of the beetle leg muscle under electrical stimulation with multiple pulse trains (100 Hz, 10% duty cycle, 2 V) for 3 s.

Pseudocolor time series images of beetle leg muscle dosed with (A) Fluo-8 (60 µM) and (C) Cell Tracker (60 µM) indicators. ROIs are indicated by yellow region (as also shown in S1B Fig.). The color scale is given on the right side of each image (A) or (C) respectively. Fluorescence intensity dynamics of (B) Fluo-8 and (D) Cell Tracker under electrical stimulation, digitized with ImageJ software from the ROI shown in (A) and (C) respectively. The stimulus timing is indicated by grey shading. The Fluo-8 pseudocolor images illustrate the fluorescence intensity dynamics that correspond to the Ca2+ dynamics inside the leg muscle: it increased at the start of electrical stimulation, was maintained during the application of the stimulus, and finally slowly decreased after the stimulus stopped. The Cell Tracker pseudocolor images display that the muscle displacement also causes intensity change during electrical stimulation, which slightly affects the Fluo-8 measurement.

Figure 3