Figure 4.

Quantitation of spinules emerging from mushroom spines over time using Fiji/ImageJ

Enhanced resolution maximum projection images depict a GFP-labeled mushroom spine and the manual tracking of spinule lengths over time using Fiji/ImageJ software. Live time-lapse imaging was acquired at Z-intervals of 20 s for a duration of 10 min. The circle quadrant tool assists in tracking of individual spinules over time at the same topographical locations. Blue arrows highlight an elongated, long-lived spinule that appears continuously at location QII in the first 13 time-points, for an estimated lifespan of 260 s. The spinule disappears and another re-emerges at the same location at a later time-point (T=360 s), in a phenomenon termed spinule recurrence. The red arrow highlights a smaller, short-lived spinule that appears at a single time-point in QIII, for an estimated lifespan of ≤20 s. Scale bar equals 1 micron.