Leica cm3050

Histological analysis and immunohistochemistry staining were performed as previously described [38 (link)]. OCT-embedded common carotid arteries were cut systematically in serial 5-μm cross sections using a cryotome (Leica CM3050S, Leica Microsystems, Germany). Analysis was carried out in the injured left common carotid artery, whereas the contralateral served as a control. For morphometric analysis, sections were stained with hematoxylin and eosin (HE). For immunofluorescence analysis, sections were stained with a rabbit anti-rat CD31 antibody (1:250, Abcam, USA) and visualized using an Alexa Fluor 647 mouse-anti-rabbit IgG secondary antibody (1:500, eBioscience). For immunohistochemistry analysis, sections were stained with a rabbit anti-rat CD31 antibody (1:150, Abcam, USA) and visualized using a mouse-anti-rabbit secondary antibody (1:500, Sigma). Negative controls using IgG controls matching in species and concentration were run in parallel. Pictures were taken with a microscope (Leica CM3050S, Leica Microsystems, Germany) and a digital camera (DFC 320, Leica Microsystems) at ×100 magnification. Morphometric analysis was performed per sample followed by computer-assisted morphometric analysis (ImageJ, NIH, USA). Subsequent morphometric analyses were performed in a double-blinded manner. Four animals per group and 3 samples per animal were analyzed.

Ungroomed fingermarks were prepared and deposited onto aluminium sheets (which were pretreated to remove the stationary phase) as previously described [15 (link)]. Fresh frozen rat brain tissue sections were cut to 10 μm thickness using a Leica CM3050 cryostat (Leica Microsystems, Milton Keynes, UK) operating at –20°C. Sections were subsequently thaw-mounted onto poly-lysine coated glass slides and stored in an airtight tube at –80°C. All animal tissue studies were carried out in accordance with Animals (Scientific Procedures) Act 1986 and the GSK Policy on the Care, Welfare, and Treatment of Laboratory Animals.

Animals were deeply anaesthetized with isoflurane and transcardially perfused with 4% formaldehyde solution. Brains from these animals were isolated, kept in 30% sucrose solution for 24 hours for cryoprotection, snap frozen in dry ice-cooled isopentane, and stored in -80°C until further processing. Next, coronal slices of the hippocampal-regions were serially sectioned at 18-μm using a microtome at -20°C (Leica CM 3050; Leica Microsystems) and mounted onto silanized glass slides. Glass slides were kept at −80°C until further use.