Cm1520

Manufactured by Leica Biosystems

Sourced in Germany

The CM1520 is a cryostat designed for sectioning frozen tissue specimens. It features a temperature range of -10°C to -35°C and an integrated refrigeration system. The instrument is capable of producing sections ranging from 1 to 300 microns in thickness.

Automatically generated - may contain errors

Lab products found in correlation

Normal saline, by Merck Group (2 mentions) Eclipse c2 plus, by Nikon (2 mentions) Paraformaldehyde (pfa), by Biosesang (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Normal goat serum (ngs), by Cell Signaling Technology (1 mentions) Image pro plus program, by Media Cybernetics (1 mentions) Model cx31rtsf, by Olympus (1 mentions) Tp1020, by Leica Biosystems (1 mentions) Eg1150h, by Leica Biosystems (1 mentions) Safranin o fast green staining, by Merck Group (1 mentions) Alexa fluor 488 labeled goat anti mouse secondary antibody, by Abcam (1 mentions) Hematoxylin and eosin, by Merck Group (1 mentions) Fitc conjugated goat anti rabbit igg antibody, by Proteintech (1 mentions) Eclipse ti, by Nikon (1 mentions)

8 protocols using cm1520

Muscle Fiber Typing by Immunofluorescence

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Transverse sections were cut into 10 μm thickness using a cryostat
(CM1520, Leica Biosystems, Wetzlar, Germany) at −20°C. The
sections were pre-incubated in 10% normal goat serum (Cell Signaling Technology,
Danvers, MA, USA) for blocking. Primary antibodies (DSHB, IA, USA) were used for
detecting one or more MHC isoforms (BA-F8, slow/I; SC-71, 2a and 2x; BF-35, all
isoforms except for 2x; 6H1, 2x; BF-F3, 2b). For multicolor immunofluorescence,
Alexa Fluor 350, 488, 594, and 647 (Thermo Fisher Scientific, Waltham, MA, USA)
were applied to each section for 1 h at room temperature. Primary and secondary
antibodies were applied serially or in a cocktail to the sections, respectively.
The dilution ratio and configuration of antibodies are presented in Table 1. All the sections were rinsed in
PBS for 5 min with triplication after incubation. The sections were visualized
with confocal scanning laser microscope (TCS SP8 STED, Leica Biosystems,
Wetzlar, Germany). Cross-sectional area (μm²), relative number
composition (%), and relative area composition (%) of each muscle fiber type
were analyzed from approximately 800 fibers per section using Image Pro Plus
program (Media Cybernetics, Rockville, MD, USA).

Song S., Ahn C.H, & Kim G.D. (2020). Muscle Fiber Typing in Bovine and Porcine Skeletal Muscles Using Immunofluorescence with Monoclonal Antibodies Specific to Myosin Heavy Chain Isoforms. Food Science of Animal Resources, 40(1), 132-144.

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Histopathological Analysis of Paw Tissue

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Paw tissue samples were removed at the 40th day post-infection, fixed in 4% formaldehyde and processed for paraffin embedding. Tissue longitudinal sections (5 μm) were prepared in cryostat (CM1520, Leica Biosystem, Richmond, IL, USA) and slides stained with hematoxylin and eosin (H&E). The analysis of the slides (four slides per mice/four animal per group) was performed using light microscopy (Olympus Life Science, model CX31RTSF, Tokyo, Japan) with magnification of × 40 on the panels j, k, and m (scale bars 20 μm) and × 100 on the panel l (scale bar 10 μm) and presented in Fig. 1. Histopathological score of epidermis and dermis thickening was also performed for the experimental groups with magnification of × 200 and presented in panel n of Fig. 1.

Borghi S.M., Fattori V., Pinho-Ribeiro F.A., Domiciano T.P., Miranda-Sapla M.M., Zaninelli T.H., Casagrande R., Pinge-Filho P., Pavanelli W.R., Alves-Filho J.C., Cunha F.Q., Cunha T.M, & Verri WA J.r. (2019). Contribution of spinal cord glial cells to L. amazonensis experimental infection-induced pain in BALB/c mice. Journal of Neuroinflammation, 16, 113.

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Comprehensive Tissue Collection Protocol

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At the end-point of each experiment animals, 200 μl of retro-orbital blood was collected from anesthetized animals and afterwards euthanasia was performed by cervical dislocation. For tissue array, necropsy of animals was performed according to the methods described by the Jackson laboratory. The order of extraction was the following according to the order of tissue autolysis: liver, pancreas, kidney, brain and muscle. Samples extracted were collected in Optimal Temperature Compound (O.C.T) and flash frozen fresh until further use. Before staining, O.C.T. included samples were cut using cryostat (Leica Biosystems CM1520). Alternatively, biological tissues were fixed in 10% neutral buffered formalin (NBF) (20:1 v/v). The 10% NBF was replaced by fresh fixative to eliminate blood and feces from the fixative. After 24h of fixation, tissues were placed in 50% alcohol and processed and embedded in paraffin with an automated tissue processor (Leica Biosystems TP1020, Barcelona, Spain) and an embedding center (Leica Biosystems EG1150H, Barcelona, Spain). Tissues were oriented properly previous to the final embedding in paraffin.

Gutiérrez-de-Juan V., López de Davalillo S., Fernández-Ramos D., Barbier-Torres L., Zubiete-Franco I., Fernández-Tussy P., Simon J., Lopitz-Otsoa F., de las Heras J., Iruzubieta P., Arias-Loste M.T., Villa E., Crespo J., Andrade R., Lucena M.I., Varela-Rey M., Lu S.C., Mato J.M., Delgado T.C, & Martínez-Chantar M.L. (2017). A morphological method for ammonia detection in liver. PLoS ONE, 12(3), e0173914.

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Golgi Staining and Dendritic Spine Analysis

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Brains were put into Golgi immersion solution (Solution A containing potassium dichromate and mercuric chloride: Solution B containing potassium chromate =1:1) at room temperature in the dark for 14 days, with the immersion solution changed each day. After two weeks, the brains were put into Solution C for 5 days, with the solution refreshed each day. Brains were placed on sample platforms and quickly frozen on dry ice for 10 min. Samples were cut into 100-μm-thick sections with a frozen section microtome (CM1520; Leica Biosystems), which were suitable for pathological and biological examination Sections were mounted on gelatinated slides and dried in the dark. Sections were washed with distilled water, stained with dye for 10 min, and washed again. The stained sections were dehydrated in 50%, 75% and 95% ethanol for 4 min and then dehydrated in anhydrous ethanol 4 times. Sections were washed with xylene 3 times and sealed with resin
[19] (link). Dendritic spines were observed with a Digital Tissue Slice Scanning System (Aperio, San Diego, USA). Dendritic spines were manually marked and total numbers of dendritic spines were calculated using Image J software (NIH, Bethesda, USA)
[19] (link).

Cai H., Qiao J., Chen S., Yang J., Hölscher C., Wang Z., Qi J, & Wu M. (2022). MCU knockdown in hippocampal neurons improves memory performance of an Alzheimer’s disease mouse model : MCU knockdown improves memory performance. Acta Biochimica et Biophysica Sinica, 54(10), 1528-1539.

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Histological Analysis of Intervertebral Disc Degeneration

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The rats were perfused transcardially with 0.9% normal saline (Sigma-Aldrich, St. Louis, MO, USA) and 4% paraformaldehyde (Biosesang, Seong-nam, Korea) for histological staining and immunohistochemistry. The L4–5 and L5–6 discs were extracted, then post-fixed in 4% paraformaldehyde overnight at 4 °C. Tissues were immersed in 30% sucrose for 3 days and sectioned in 20-µm thicknesses using cryo-microtome (CM1520, Leica Biosystems, Nussloch, Germany). Safranin O/fast green staining (Sigma-Aldrich, St. Louis, MO, USA) was performed according to the manufacturer’s instructions on the L4–5 and L5–6 discs to confirm the degree of damage to the disc at 4 weeks. Stained sections were imaged using an inverted microscope (Eclipse C2 Plus, Nikon). The damage grade in the intervertebral discs was assessed using a histological grading scale based on four degenerative change categories (to assess the anulus fibrosus, the border between the anulus fibrosus and nucleus pulposus, the cellularity of the nucleus pulposus, and the matrix of the nucleus pulposus), with scores ranging from a normal disc with 4 points (1 point in each category) to a severely degenerated disc with 12 points (3 points in each category) [16 (link)].

Hong J.Y., Kim H., Jeon W.J., Lee J., Yeo C., Lee Y.J, & Ha I.H. (2022). Epigenetic Changes within the Annulus Fibrosus by DNA Methylation in Rat Intervertebral Disc Degeneration Model. Cells, 11(22), 3547.

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Spinal Cord Macrophage Immunostaining

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Transcardiac perfusion with 0.9% normal saline (Sigma-Aldrich, St. Louis, MO, USA) and 4% paraformaldehyde (Biosesang, Seongnam, Republic of Korea) was performed 3 weeks after catheter implantation in lumbar spinal stenosis (LSS) rats. The 2 cm spinal cord sample from the T10 level (start of catheter insertion) was post-fixed overnight in 4% paraformaldehyde at 4 °C and cryoprotected in 30% sucrose in PBS for 3 days. Tissues were sectioned in the sagittal plane into 16 µm sections using a cryo-microtome (CM1520, Leica Biosystems, Nussloch, Germany). The sectioned tissues were incubated with rabbit anti-monocytes/macrophages (1:500; Abcam, Cambridge, UK) at 4 °C overnight. After washing thrice with PBS, Alexa Fluor 488-labeled goat anti-mouse secondary antibody (Abcam) was added at 1:200 in 2% normal goat serum for 2 h. The stained tissue was imaged using a confocal microscope (Eclipse C2 Plus, Nikon, Tokyo, Japan).

Hong J.Y., Kim H., Lee J., Jeon W.J., Yeo C., Kim H., Lee Y.J, & Ha I.H. (2023). Epidural Injection Method for Long-Term Pain Management in Rats with Spinal Stenosis. Biomedicines, 11(5), 1390.

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Histological Analysis of OVA-Coated ZMNs in Mouse Skin

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The excised
BALB/c mouse abdominal
skin was mounted on a polystyrene block, and OVA-coated ZMNs were
inserted in the skin for 30 min. Then, the skin sample was embedded
in an optimum cutting temperature (OCT) compound at −80 °C.
The skin sample was sectioned at a thickness of 6 μm using a
cryostat (CM1520, Leica Biosystems, Germany). Later, the skin sections
were dried overnight and stained with hematoxylin and eosin (Sigma-Aldrich).
Microscopic images were acquired using an optical microscope.

Bhatnagar S., Chawla S.R., Kulkarni O.P, & Venuganti V.V. (2017). Zein Microneedles for Transcutaneous Vaccine Delivery: Fabrication, Characterization, and in Vivo Evaluation Using Ovalbumin as the Model Antigen. ACS Omega, 2(4), 1321-1332.

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Immunofluorescence Analysis of CCR1 Expression

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The sections were cut with 4 μm thickness from frozen endobronchial biopsies using a freezing microtome (CM1520; Leica Biosystems, Shanghai, China) and kept at room temperature for 30 min. The sections were then washed with PBS for 5 min three times, incubated for 5-10 min in 3% H₂O₂ to eliminate endogenous peroxidase activity, followed by washing with PBS for 5 min twice, and incubated for 1 h with a blocking solution (10% goat serum). Next, the sections were incubated for 30 min with rabbit polyclonal anti-CCR1 antibody, then incubated with FITC-conjugated goat anti-rabbit IgG antibody (1:500; Proteintech, Rosemont, IL, USA) for 30 min at 37°C. Following nuclear staining with DAPI (1:1000; Thermo Fisher Scientific) the sections were observed and analyzed using a fluorescence microscope (Nikon Eclipse TI; Nikon, Tokyo, Japan).

Zhao K., Dong R., Yu Y., Tu C., Li Y., Cui Y., Bao L, & Ling C. (2020). Cigarette smoke-induced lung inflammation in COPD mediated via CCR1/JAK/STAT /NF-κB pathway. Aging (Albany NY), 12(10), 9125-9138.

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