Dear Elodie!

We have done several sets of IHC on 4µm mouse brain sections using the Leica Bond III system (Leica Biosystems Newcastle Ltd, UK).

The paraffin-embedded tissue blocks were chilled on ice and sliced on a Leica (Leica Biosystems, Nussloch, Germany) RM2235 manual rotary microtome at a thickness of 4 μm. The sections were mounted onto gelatin-coated slides, dried overnight at room temperature, and stored at 4 °C inside storage boxes.

Tissues were pretreated with epitope-retrieval solutions (ER, Leica Biosystems Newcastle Ltd, UK) followed by 30 minutes incubation with primary antibodies.

We used ARGI (GeneTex, GTX113131), CD68 (Abcam, # ab125212) and ARGII (GeneTex, #GTX104036) Abs with dilutions 1:500, 1:200, and 1:600, respectively. ER/time 2/20 (min)

Of note, IBA1 Ab was used as a standard marker for microglia.

The Leica Refine-HRP kit (Leica Biosystems Newcastle Ltd, UK) used for detection and counterstain with Hematoxylin.

Best regards

https://link.springer.com/article/10.1007/s13311-018-0669-5

https://www.mdpi.com/1422-0067/20/18/4616/htm

Thanks a lot for your answer. Really useful!