Figure 3.

Nanoscale association of β/γ-secretase and with APP in the neuronal processes using STED microscopy

(A and D) Gallery of confocal images of neuronal processes identified by automatic detection of confocal marker for APP (magenta) puncta with pseudocolour overlay of β/γ-secretase (green).

(B and E) STED image of the respective regions identified from (A, D).

(C and F) Represent automatically detected regions for confocal marker for β/γ-secretase (black) along with nanoscale representation of β/γ-secretase (green) and APP (magenta). Black in the overlay images represents the overlap between the corresponding green and magenta images.

(i and iii) Indicate the distribution of length of all β/γ-secretase nanodomains on APP obtained by STED microscopy.

(ii and iv) Indicate the distribution of intensity of all β/γ-secretase nanodomains on APP obtained by STED microscopy.

(G) Confocal image of the individual γ-secretase identified by automatic detection of confocal marker for γ-secretase (red) with pseudocolour overlay of β-secretase (blue).

(H) STED image of the representative regions from (G).

(I) Indicates the automatically detected regions for confocal marker for β-secretase (black), nanoscale representation of β-secretase (blue) and γ-secretase (red).

(v and vi) Indicate the distribution of length and intensity of all β-secretase nanodomains on γ-secretase obtained by STED microscopy, respectively. n = 13,484 puncta (β-secretase on APP), 6033 puncta (γ-secretase on APP), and 4762 puncta (for β/γ-secretases). Scale bar at (C, F, I) indicates 1.1 μm.