Immunohistochemical staining of fixed-frozen sections (40 μm thickness) was performed as described previously [28 (link)]. Briefly, the brain slices were mounted onto gelatin-coated slides, and primary antibody (rabbit anti-Iba1, Wako, 1:250) was added overnight. Secondary antibodies (1:1000) with corresponding conjugates and 4′,6-diamidino-2-phenylindole and dihydrochloride (DAPI, 1:1000, Invitrogen, Carlsbad, CA) were applied. Images were analyzed with a Zeiss Axiovert 200M microscope (Carl Zeiss, Oberkochen, Germany) and ImageJ software (NIH).
4 6 diamidino 2 phenylindole and dihydrochloride dapi
Manufactured by Thermo Fisher Scientific
4',6-diamidino-2-phenylindole and dihydrochloride (DAPI) is a fluorescent stain that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used in molecular biology and microscopy applications.
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2 protocols using 4 6 diamidino 2 phenylindole and dihydrochloride dapi
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Immunohistochemical Staining of Brain Slices
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Immunohistochemical Staining of Brain Slices
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Immunohistochemical staining of fixed-frozen sections (30 μm thickness) was performed as described previously (Ritzel et al., 2019 (link)). Briefly, the brain slices were mounted onto gelatin-coated slides, and primary goat anti-CD200R (ThermoFisher Scientific, cat # PA5–47345) and rabbit anti-VWF (ThermoFisher Scientific, cat # 27186–1AP) were used for staining. Brain slices were further stained with secondary antibodies conjugated to Alexa Fluor 594, and Alexa Fluor 488 (1:400), and followed with 4′,6-diamidino-2-phenylindole and dihydrochloride (DAPI, 1:1000, Invitrogen, Carlsbad, CA), respectively. Images were analyzed with a Zeiss Axiovert 200 M microscope (Carl Zeiss, Oberko-chen, Germany) and ImageJ software (NIH).
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