Microm hm 525 cryostat

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany

The Microm HM 525 Cryostat is a laboratory equipment designed for the preparation of frozen tissue sections. It provides a controlled low-temperature environment for sectioning samples, enabling the preservation of tissue morphology for further analysis.

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Lab products found in correlation

Lsm 780 confocal microscope, by Zeiss (6 mentions) Axio observer z1, by Zeiss (3 mentions) Cd11b bv421, by BD (2 mentions) Fluoromount g mounting medium, by Thermo Fisher Scientific (2 mentions) Clear oct compound, by Thermo Fisher Scientific (2 mentions) Anti hamster af488 conjugate, by BioLegend (2 mentions) Igm af568, by Thermo Fisher Scientific (2 mentions) Fisherbrand superfrost plus microscope slide, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594 conjugate, by Cell Signaling Technology (2 mentions) Rabbit anti mouse iba 1 primary antibody, by Fujifilm (2 mentions) Vectashield dapi 4 6 diamidino 2 phenylindole 2hcl, by Vector Laboratories (2 mentions) Goat anti rabbit igg h l secondary antibody, by Vector Laboratories (2 mentions) Superfrost plus slides, by Avantor (2 mentions) Superfrost plus, by Thermo Fisher Scientific (2 mentions) Oil red o, by Sangon (2 mentions) Oil red o, by Merck Group (1 mentions) Tissue tek oct compound, by Sakura Finetek (1 mentions) Sprayer, by HTX Technologies (1 mentions) Filter paper, by Merck Group (1 mentions) Rlt buffer, by Qiagen (1 mentions)

Close spelling variants of this product

Microm HM 525 HM 525 Cryostat Microm cryostat Cryostat

25 protocols using microm hm 525 cryostat

Cardiac Lipid and Neutrophil Analysis

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Hearts were harvested, fresh-frozen in OCT Tissue Tec (Sakura® Finetek), and cut into 6 μm sections (Microm HM 525 Cryostat, Thermo Fisher Scientific). Cardiac sections were stained to detect neutral lipid content using Oil Red O (ORO, Sigma-Aldrich). To analyze neutrophils, tissue sections were stained using rat anti-mouse Ly6G (Gr-1) monoclonal antibody (ebioscience), followed by secondary Alexa Fluor 546–labeled anti-rat antibody (Life Technologies), and Hoechst 33342 dye counterstaining (Thermo Fisher Scientific). More details are given in the online-only Data Supplement.

Wallert M., Ziegler M., Wang X., Maluenda A., Xu X., Yap M.L., Witt R., Giles C., Kluge S., Hortmann M., Zhang J., Meikle P., Lorkowski S, & Peter K. (2019). α-Tocopherol preserves cardiac function by reducing oxidative stress and inflammation in ischemia/reperfusion injury. Redox Biology, 26, 101292.

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Immunofluorescent Imaging of Leishmania Infection

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Spleens of naive and infected C57BL/6 mice were collected on days 21 and 28 postinfection and snap-frozen in blocks of clear OCT compound (Fisher Health Care) using liquid nitrogen. Blocks were stored at −80°C until being sectioned into 10-μm slices using a Microm HM525 cryostat (Thermo Fisher) at −0°C. Tissue sections were mounted on glass slides and immediately processed by air drying and rehydrating tissues in PBS for 30 min. Unspecific staining was blocked by treatment of tissues with PBS containing 5% BSA and 1:100 of FcBlock antibody (2.4G2) for 1 h. LV9 amastigotes were stained by incubating tissues with serum from LV9-infected hamsters at a concentration of 1:500 and then labeling bound anti-LV9 using an anti-hamster-AF488 conjugate (BioLegend). Tissue cells were labeled using CD11b-BV421 (BD Biosciences) and IgM-AF568 (Invitrogen) and subsequently fixed using 1% paraformaldehyde (PFA) before mounting coverslips on Fluoromount-G mounting medium (Invitrogen) over the tissue. Images were taken on an LSM780 confocal microscope (Zeiss) using either a 40× or 63× oil immersion objective.

Stögerer T., Silva-Barrios S., Carmona-Pérez L., Swaminathan S., Mai L.T., Leroux L.P., Jaramillo M., Descoteaux A, & Stäger S. (2023). Leishmania donovani Exploits Tunneling Nanotubes for Dissemination and Propagation of B Cell Activation. Microbiology Spectrum, 11(4), e05096-22.

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Mouse Skin Sectioning and Preservation

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Glabrous skin from the hindpaw and hairy skin from the back of the mouse were depilated with Surgi Cream-Extra Gentle for Face (Ardell, Los Angeles, CA, USA) and removed by blunt-dissection. Skin pieces were spread on an index card with the epidermal surface up, fixed in 4% PFA for 20 minutes, washed three times in phosphate-buffered saline (PBS) for 5 minutes/wash, then sunk in 30% sucrose in PBS for 24h. The skin was removed, embedded in Tissue-Tek OCT compound (Sakura Finetek, Torrance, CA, USA), and frozen. Samples were cut in a cross-sectional plane at 25μm thickness using a Microm™ HM 525 Cryostat (Thermo Scientific, Waltham, MA, USA) and were thaw mounted onto Fisherbrand Superfrost Plus microscope slides (Fisher Scientific, Pittsburgh, PA, USA). Slides with sections were kept at −80°C until use.

Cabañero D., Irie T., Celorrio M., Trousdale C., Owens D.M., Virley D., Albrecht P.J., Caterina M.J., Rice F.L, & Morón J.A. (2016). Identification of an epidermal keratinocyte AMPA glutamate receptor involved in dermatopathies associated with sensory abnormalities. Pain Reports, 1(3), e573.

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SR-FTIR Analysis of Rice Leaf Samples

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The rice leaf samples were thinned to 7 μM before being scanned for SR-FTIR microspectroscopy. In all treatments, rice leaf samples were collected 7 days after the Xoo challenge. They were cut into 4 × 8 mM pieces and immediately coated with the Tissue-Trek optimum cutting temperature (OCT) compound (Electron Microscopy Sciences Inc., Hatfield, USA) and frozen. Until cryosectioning, each sample was kept at −80 °C. Through use of a Microm HM 525 Cryostat (Thermo Fisher Scientific), the sample was cut into small pieces with a thickness of 7 μM and the full leaf structure (epidermis and mesophyll) was preserved. The frozen rice leaves were placed on a BaF2 window, which is an infrared transparent material. To avoid the influence of water peaks in the examination, the water moisture in the sample was evaporated using a desiccator. The SR-FTIR spectra were chosen to be in the mid-infrared range [26 (link)].

Siriwong S., Thepbandit W., Hoang N.H., Papathoti N.K., Teeranitayatarn K., Saardngen T., Thumanu K., Bhavaniramya S., Baskaralingam V., Le Thanh T., Phansak P, & Buensanteai N. (2021). Identification of a Chitooligosaccharide Mechanism against Bacterial Leaf Blight on Rice by In Vitro and In Silico Studies. International Journal of Molecular Sciences, 22(15), 7990.

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Cryosectioning and MALDI Analysis of Root Nodules

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Root nodules from control and high salt plants were trimmed from the plants with 2–4 mm of the surrounding root. Nodules were embedded in 100 mg/mL gelatin and frozen on dry ice. Nodules were sectioned at 16 um thickness on a Microm HM 525 cryostat (Thermo Fisher Scientific) at -20°C. Sections were thaw-mounted onto plain glass microscope slides for analysis on the MALDI LTQ Orbitrap XL or indium tin oxide coated glass slides for analysis on the AP-MALDI QE-HF system. A TM Sprayer (HTX Technologies, LLC, Carrboro, NC, United States) was used to apply DHB and CHCA matrix. DHB matrix (40 mg/mL in 50% methanol, 0.1% formic acid) was applied with a 24 pass TM Sprayer method (30 s dry time in between passes, 90° rotation between passes and the spacing offset in between every two passes, 3 mm spacing, 1250 velocity, 80°C temperature, and 0.05 mL/min flow rate). CHCA matrix (10 mg/mL in 70% acetonitrile, 0.1% formic acid) was applied with a 4 pass TM Sprayer method (30 s dry time in between passes, 90° rotation between passes and the spacing offset in between every two passes, 1.5 mm spacing, 1200 velocity, 75°C temperature, and 0.24 mL/min flow rate). Matrix covered samples were stored in a dry box at -20°C until analysis.

Keller C., Maeda J., Jayaraman D., Chakraborty S., Sussman M.R., Harris J.M., Ané J.M, & Li L. (2018). Comparison of Vacuum MALDI and AP-MALDI Platforms for the Mass Spectrometry Imaging of Metabolites Involved in Salt Stress in Medicago truncatula. Frontiers in Plant Science, 9, 1238.

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Preparation of Mouse Retinal Cryosections and Whole-mounts

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Mice (28–60 days old; male) were euthanized using carbon dioxide and pneumothorax. Eyes were removed and placed in cooled, oxygenated dissecting HEPES-buffered solution containing (in mM): 137 NaCl, 2.5 KCl, 2.5 CaCl₂, 1.0 MgCl₂, 10 HEPES, 28 glucose, adjusted to pH 7.4 with NaOH. Using a dissecting stereomicroscope, the cornea and lens were quickly removed to obtain the eye-cup. For some experiments, the retina was marked to identify dorsal and ventral sides. The eyecups were fixed using 4% paraformaldehyde in 0.1 M PB, pH 7.4, for 30 min at room temperature. After fixation, retinas were separated from the sclera, rinsed several times in 0.1 M PB, and cryoprotected in 30% sucrose overnight at 4 °C. The tissue was embedded in Tissue Freezing Medium (EMS) and was sectioned vertically at 14–16 µm with a Microm HM 525 cryostat (Thermo Fisher Scientific, Waltham, MA). Cryosections were collected on histo-bond slides (Fisher Scientific). For whole-mount retinal preparation, the retina was isolated from the eye-cup in oxygenated HEPES-buffered solution, and was flattened on filter paper (EMD Millipore, Billerica, MA). Then, the tissue was fixed using 4% paraformaldehyde for 30 min, washed with 0.1M PB, pH 7.4 and processed for free-floating immunohistochemistry.

Farshi P., Fyk-Kolodziej B., Krolewski D.M., Walker P.D, & Ichinose T. (2015). Dopamine D1 receptor expression is bipolar cell type-specific in the mouse retina. The Journal of comparative neurology, 524(10), 2059-2079.

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Immunohistochemical Analysis of Microglia in Mouse Brain

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Mouse brain was fixed in 4% formaldehyde for 24 h, and then incubated in 30% sucrose until tissues are sink. Fixed brain was flash frozen using pre-cooled isopentane (− 78 °C), sectioned at 30 μm using Microm HM525 Cryostat (Thermo) and picked up on Superfrost Plus slides (VWR, 48311-703). Sections were blocked with 5% normal goat serum and washed in PBS with 1% bovine serum albumin (BSA) and incubated with rabbit- anti-mouse Iba-1 primary antibody (FUJIFILM Wako Pure Chemical Corporation 019-19741, 1:500) or rabbit- anti-mouse Iba-1 primary antibody Alexa Fluor^® 594 conjugate (Cell Signaling, 48934, 1:50) overnight. Sections were washed with phosphate buffer with 1% Tween 20 (PBS-T), and then incubated in goat anti-rabbit IgG (H + L) secondary antibody (Vector laboratory CY-1300, 1:250) at room temperature for 1 h when unconjugated antibody was used. Afterword, sections were washed three times with PBS-T followed by mounting on coverslip using Vectashield DAPI (4′6-diamidino-2-phenylindole 2HCl, Vector Labs, Burlingame, U.S.) mounting media to detect nuclei.

Al Omran A.J., Shao A.S., Watanabe S., Zhang Z., Zhang J., Xue C., Watanabe J., Davies D.L., Shao X.M, & Liang J. (2022). Social isolation induces neuroinflammation and microglia overactivation, while dihydromyricetin prevents and improves them. Journal of Neuroinflammation, 19, 2.

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Immunohistochemical Analysis of Microglial Activation

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Mouse brain was fixed in 4% formaldehyde for 24 hours, and then incubated in 30% sucrose until tissues are sink. Fixed brain was flash frozen using pre-cooled isopentane (−78 °C), sectioned at 30 μm using Microm HM525 Cryostat (Thermo) and picked up on Superfrost Plus slides (VWR, 48311–703). Sections were blocked with 5% normal goat serum and washed in PBS with 1% bovine serum albumin (BSA) and incubated with rabbit- anti-mouse Iba-1 primary antibody (FUJIFILM Wako Pure Chemical Corporation 019–19741, 1:500) or rabbit- anti-mouse Iba-1 primary antibody Alexa Fluor^® 594 conjugate (Cell Signaling, 48934, 1:50) overnight. Sections were washed with phosphate buffer with 1% Tween 20 (PBS-T), and then incubated in goat anti-rabbit IgG (H+L) secondary antibody (Vector laboratory CY-1300, 1:250) at room temperature for 1 hour when unconjugated antibody was used. Afterword, sections were washed three times with PBS-T followed by mounting on coverslip using Vectashield DAPI (4′6-diamidino-2-phenylindole 2HCl, Vector Labs, Burlingame, U.S.) mounting media to detect nuclei.

Al Omran A.J., Shao A.S., Watanabe S., Zhang Z., Zhang J., Xue C., Watanabe J., Davies D.L., Shao X.M, & Liang J. (2021). Social Isolation Induces Neuroinflammation And Microglia Overactivation, While Dihydromyricetin Prevents And Improves Them. Research Square.

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Corneal Histology and Gene Expression

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Out of ten animals in each group, corneas from five animals were used for histology and immunofluorescence staining, and the remaining five animals were used for quantitative real-time polymerase chain reaction (qRT-PCR). Eyes were enucleated using sharp Westcott scissors and 0.12 forceps, embedded in optimal cutting temperature (OCT) compound (Sakura FineTek, Torrance, CA) within a 15 mm × 15 mm × 5 mm mold (Fisher, Pittsburgh, PA) and snap-frozen. The frozen tissue blocks were stored at − 80 °C until sectioning. Tissues were sectioned at 8 μm thickness using a Microm HM525 cryostat (Thermo Fisher Scientific, Waltham, MA) and mounted on microscopic glass slides (Superfrost Plus, Fisher Scientific, Pittsburgh, PA, USA) and stored at − 80 °C until hematoxylin and eosin staining and immunofluorescence staining were performed (Mohan et al., 2008 (link)). For qRT-PCR, corneas were dissected from the enucleated eyeballs, placed in 300 μL of RLT buffer (Qiagen, Valencia, CA), and stored at − 80 °C until RNA extraction.

Balne P.K., Gupta S., Zhang J., Bristow D., Faubion M., Heil S.D., Sinha P.R., Green S.L., Iozzo R.V, & Mohan R.R. (2021). The functional role of decorin in corneal neovascularization in vivo. Experimental eye research, 207, 108610.

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Immunofluorescent Imaging of Leishmania Infection

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