The SIM on brain sections was done with a Nikon SIM system with a 100 × 1.49 NA oil immersion objective, managed by NIS elements software. Tissues were imaged at laser excitation of 488 (for Tmem119) and 640 nm (for Iba1) with a 3D-SIM acquisition protocol. Fourteen-bit images sized 1024 × 1024 pixels, with a single-pixel of 0.030 μm, were acquired in a gray level range of 0–4,000 to exploit the linear range of the camera (iXon ultra DU-897U, Andor) and to avoid saturation. Raw and reconstructed images were verified by the SIMcheck ImageJ plugin (Ball et al., 2015 (link)).
Mercurio D., Fumagalli S., Schafer M.K., Pedragosa J., Ngassam L.D., Wilhelmi V., Winterberg S., Planas A.M., Weihe E, & De Simoni M.G. (2022). Protein Expression of the Microglial Marker Tmem119 Decreases in Association With Morphological Changes and Location in a Mouse Model of Traumatic Brain Injury. Frontiers in Cellular Neuroscience, 16, 820127.
Corresponding Organization : Mario Negri Institute for Pharmacological Research
Other organizations :
Philipps University of Marburg, University of Giessen, Consejo Superior de Investigaciones Científicas, Institut d'Investigacions Biomèdiques de Barcelona, Consorci Institut D'Investigacions Biomediques August Pi I Sunyer
Nikon SIM system with a 100 × 1.49 NA oil immersion objective
NIS elements software
3D-SIM acquisition protocol
Fourteen-bit images sized 1024 × 1024 pixels
Single-pixel of 0.030 μm
Gray level range of 0–4,000
IXon ultra DU-897U camera (Andor)
controls
Raw and reconstructed images were verified by the SIMcheck ImageJ plugin (Ball et al., 2015)
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