Immunostaining was adapted from Micheva and Smith1 with modifications. Instead of a PAP pen, we used reversibly applicable and removable custom sample chambers made of polydimethylsiloxane (PDMS, Sylgard 184, Dow Corning) for IHC and dSTORM image acquisition. According to the instruction for use, the bulk material is thoroughly mixed with the crosslinker compound, degassed, and subsequently polymerized at 85°C for 4 h in negative molds made from Plexiglas. The resultant rectangular chambers with an inner edge length of 1 cm reversibly adhere to glass slides, creating a barrier for keeping IHC or switching buffers in place (cf. Fig. 1). For initial blocking and rehydration, blocking solution (0.1% BSA and 0.05% Tween 20 in Tris-buffer) was added for 5 min. All incubation steps were performed in humidity chambers at room temperature. The first antibodies were diluted (monoclonal mouse anti--tubulin, , Sigma; product number: T5168) or (polyclonal chicken anti-GFP, abcam; product number: ab13970) in blocking solution, centrifuged for 2 min at top speed and applied for 1 h. Sections were washed four times with Tris-buffer (5 min each). Secondary antibodies [for dSTORM application: goat anti-mouse IgG (H+L) Alexa Fluor 647 conjugate, F(ab')2 fragment, Thermo Fisher Scientific; for SIM application: goat anti-chicken IgG (H+L) Alexa Fluor 488 conjugate, Thermo Fisher Scientific, and goat anti-mouse IgG (H+L) Texas Red dye-conjugate, Jackson ImmunoResearch] were also diluted in blocking solution and then applied for 30 min in the dark. After washing Live Hoechst 33342 (Sigma) diluted in Tris-buffer was applied, then sections were washed again two times, and finally either PDMS sample chambers were removed and the sections mounted with Mowiol (for SIM application) or sections were kept in Tris-buffer until dSTORM application. All light microscopic analyses were performed within 3 days of immunostaining. SIM image acquisition was performed with an ELYRA S.1 super-resolution structured illumination microscope (Zeiss). Black glue in between sections and the Hoechst staining aided in finding the sections and tissues.