All samples for LCM were cut with a microtome to 10 µm slices (Leica, RM2055) and were mounted on glass slides (SuperFrost Ultra Plus, Menzel Gläser, Thermofisher Scientific [33 (link)]) with a drop of DNAse/RNAse-free water. Next, samples were incubated in a fume hood at 56 °C for 1 h to increase adherence to slides. Mounted slices were hematoxylin stained according to the standard protocol in a set of alcohol solutions, xylene, and stain.
Rm 2055
Manufactured by Leica camera
The Leica RM 2055 is a rotary microtome designed for histological and cytological sample preparation. It features a robust, durable construction and precise specimen feed and sectioning mechanisms. The RM 2055 allows for the creation of thin, uniform sections from paraffin-embedded tissue samples.
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Lab products found in correlation
4 protocols using rm 2055
1
Tissue Preparation for Laser Capture Microdissection
2
Aortic Morphology Characterization
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The first aortic ring was fixed in formaldehyde 4% to preserve cellular morphology and avoid tissue degeneration; then, samples were paraffin-embedded and sectioned in 5 μm slices by microtome Leica RM 2055. After the rehydration of the sections, eosin/hematoxylin staining was performed to color nucleus and cytoplasm. Finally, the colored sections were observed using microscope Nikon Ni-e (Nikon Instruments Spa, Calenzano, Italy), scanned by Nano Zoomer Hamamatsu and analyzed by software Aperio Imagescope version 12.3.3 (Leica Biosystem, Buccinasco, Italy).
For each transversal section, 40 measurements of tunica media thickness have been recorded for each aortic ring.
For each transversal section, 40 measurements of tunica media thickness have been recorded for each aortic ring.
3
Immunological Profiles in Severe Shigellosis
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Adult patients clinically suspected of having dysentery within 3–4 d of onset of disease were initially enrolled at the International Center for Diarrheal Diseases Research, Bangladesh (ICDDR,B). The collection of biopsies from patients for studying immunological/immunopathological responses was approved by the Ethical Review Committee of ICDDR,B. Patients were diagnosed for shigellosis after overnight stool culture on McKonkey and Shigella Salmonella (SS) agar plates. Confirmation of diagnosis was achieved by testing for Shigella spp. by agglutination to specific antisera and further biochemical tests. Proctoscopy (long-speculum Anoscope; Welch Allyn series 31610) was performed to obtain rectal biopsy specimens from each patient within 48 h of admission at ∼10–12 cm from the anus into the rectum. Formalin-fixed, paraffin-embedded tissues were sectioned at 3-µm thickness with a microtome (RM 2055; Leica) and stored at room temperature until analysis. Biopsies from 12 patients were analyzed by immunofluorescence. Image representation was focused on patients presenting severe shigellosis (n = 4) in whose biopsies we could identify a lymphoid follicle (n = 2).
4
Scratch Testing of Material Surfaces
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The scratch tests were carried out at VTT Technical Research Centre of Finland Ltd.
To prepare a smooth surface for scratching, a polishing technique was used by cutting a thin slice of the specimen with a microtome (model Leica Jung RM 2055). The CSM A Rockwell C diamond tip with a radius of 200 µm and a steel ball with a radius of 1000 µm were used for scratch testing, and two loading modes, constant and continuously increasing load, were adopted. The constant scratch load of 2 N and increasing load from 0 to 2 N were used for the diamond tip. For the steel ball tip, a constant scratch load of 4 N and increasing load from 0 to 4 N were adopted. Scratching velocity of 10 mm/min and the total scratch length of 10 mm were used with the two tip geometries. Between the scratches, there was a space of 1.5 mm when using the diamond tip and 2 mm when using the steel ball. Pre-scan of the scratch path was done prior to scratching using 0.1 N force to obtain the surface profile as a baseline for penetration depth and residual depth measurements.
During scratching, frictional force was measured and the related friction coefficient was determined as well as the penetration depth of the tip, which describes the combined elastic and plastic deformation. After scratching, another scan was made using 0.1 N normal force to measure the residual depth of the scratch channel.
To prepare a smooth surface for scratching, a polishing technique was used by cutting a thin slice of the specimen with a microtome (model Leica Jung RM 2055). The CSM A Rockwell C diamond tip with a radius of 200 µm and a steel ball with a radius of 1000 µm were used for scratch testing, and two loading modes, constant and continuously increasing load, were adopted. The constant scratch load of 2 N and increasing load from 0 to 2 N were used for the diamond tip. For the steel ball tip, a constant scratch load of 4 N and increasing load from 0 to 4 N were adopted. Scratching velocity of 10 mm/min and the total scratch length of 10 mm were used with the two tip geometries. Between the scratches, there was a space of 1.5 mm when using the diamond tip and 2 mm when using the steel ball. Pre-scan of the scratch path was done prior to scratching using 0.1 N force to obtain the surface profile as a baseline for penetration depth and residual depth measurements.
During scratching, frictional force was measured and the related friction coefficient was determined as well as the penetration depth of the tip, which describes the combined elastic and plastic deformation. After scratching, another scan was made using 0.1 N normal force to measure the residual depth of the scratch channel.
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