Prior to paraffin embedment, the cerebrum was sectioned in five equal pieces cut through the frontal plane at +10, +5, 0, and – 5 mm from Bregma on the anterior posterior axis. The paraffin blocks were cooled to – 10°C to ease sectioning. With the Microm HM340 E microtome, strips of nine 4 μm sections were made after which 400 μm was discarded. Eighteen strips of each brain were made for analysis of which most were close to the prefrontal and sensorimotor cortex injection sites as the parietal cortices did not show traces of an injection spot during necropsy nor with the MRI. The strips were stored at 4°C until they were separately transferred to microscope slides by the use of a water bath at 37°C (Menzel-Gläser polysine slides, Thermo Fisher scientific, Waltham MA). The slides were dried o/n at 40°C after which they were stored at roomtemperature.
Menzel gl ser polysine slides
Manufactured by Thermo Fisher Scientific
Menzel-Gläser polysine slides are glass microscope slides coated with a polysine adhesive. The slides provide a positively charged surface to enhance the adhesion of tissue sections and cell samples during mounting and staining procedures.
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Lab products found in correlation
2 protocols using menzel gl ser polysine slides
1
Sectioning and Preparation of Brain Tissue
2
Immunofluorescence Imaging of HEK293 Cells and PBMCs
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HEK293 adherent cells were seeded onto eight-well chamber slides (Thermo Scientific™ Nunc™ Lab-Tek™ II Chamber Slide™) and allowed to adhere to the slide (16 hours) before they were fixed with 4% paraformaldehyde or cold (−20°C) methanol. The cells after PBS (Biochrom, Berlin, Germany) washing, were mounted, counterstained with DAPI, and visualized under Nikon Ti confocal microscope (Sendai Nikon Corporation, Miyagi, Japan).
Nucleofected PBMCs grown in suspension were collected, fixed for 20 min in PBS containing 4% parafomaldehyde. The cells were then centrifuged, washed in deionized water and resuspended in 200 μl deionized water. The cells were smeared on adhesion slides (Menzel-Gläser Polysine Slides, Thermo Scientific), dried and washed with water for eliminating the crystals. The cells were then once again washed in PBS, mounted, stained with DAPI and examined by confocal microscopy.
Nucleofected PBMCs grown in suspension were collected, fixed for 20 min in PBS containing 4% parafomaldehyde. The cells were then centrifuged, washed in deionized water and resuspended in 200 μl deionized water. The cells were smeared on adhesion slides (Menzel-Gläser Polysine Slides, Thermo Scientific), dried and washed with water for eliminating the crystals. The cells were then once again washed in PBS, mounted, stained with DAPI and examined by confocal microscopy.
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