FIGURE 3.

Micro-scale imaging example for using SamuROI on a dendritic structure. (A) (A1) The morphological image of a dendritic segment from an electroporated layer 2 cell in the MEC. (A2) The corresponding FrameView image from SamuROI, a composite of the thresholding overlay mask defining relevant pixels of the grayscale morphological image and color-coded frame specific brightness values of the relevant pixels. The yellow outline was generated using the BranchMask tool. The scale bar corresponds to 2 μm. (A3) Depicts how the BranchMask from (A2) is divided into SegmentMasks by SamuROI. (B) RasterView of segments from the dendritic branch corresponding to (A). Time on the x-axis is defined by the scale bar in (C). ΔF/F is color-coded as in (A4). The magenta arrow corresponds to the point on the y-axis representing the magenta-colored segment both in (A4) and in (C). The black arrow corresponds to the time point indicated by arrows in (C) and (D) and defines the frame depicted in (A4). (C) The inset on the left magnifies the distal part of the dendrite shown in (A). The trace corresponds to the SegmentMask outlined in the inset and (A4). (D) Based on the distribution of pixel brightness at the time point defined by the black arrow in (B), ROI masks are defined manually as freehand PolygonMasks. The local Ca²⁺ transient in the spine (green) can be differentiated from the dendritic segment (red).