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Fig. 5 | Biological Procedures Online

Fig. 5

From: High-Resolution Confocal Fluorescence Imaging of Serine Hydrolase Activity in Cryosections – Application to Glioma Brain Unveils Activity Hotspots Originating from Tumor-Associated Neutrophils

Fig. 5

Confocal imaging of SH activity in relation to neutrophil elastase (NE). Sections went through the tissue-ABPP protocol to label SHs (red) and were thereafter immunostained for NE (yellow), followed by DAPI staining to visualize nuclei (blue). Panel a shows overall staining pattern throughout the coronal section plane. A control section undergoing identical staining protocol with no primary antibody was not available for this experiment. Panel b shows staining pattern in glioma region characterized by intense SH activity originating from individual cells (TAMRA-FP hotspots). Panel c shows staining pattern in glioma region characterized by intense SH activity originating from cell clusters (TAMRA-FP hotspot clusters). Panel d shows staining pattern in healthy brain (cortex). Note detectable expression of NE throughout the glioma (a). Note in particular NE-positive cells in the region of TAMRA-FP hotspots and close match of NE-positive cells with the TAMRA-FP signal (b). Note strong NE-immunostaining of the TAMRA-FP hotspot clusters and close match of NE staining within the TAMRA-FP hotspot cluster (c). NE is not visible in control cortical region (d). Primary antibody rabbit anti-NE (Abcam, cat# ab21595), dilution 1:500, secondary antibody Goat anti-rabbit IgG-Alexa Fluor 647 conjugate, dilution 1:100. Sections were from female rat 11. Scale bars: 1 mm in a, 20 μm in b-d. Images were adjusted for brightness and contrast

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