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Microsystems microscope

Manufactured by Leica
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The Leica Microsystems microscope is a precision optical instrument designed for detailed observation and analysis of microscopic specimens. It features high-quality optics, advanced imaging capabilities, and customizable configurations to accommodate a wide range of applications.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Image pro plus 6, by Leica (2 mentions) Sm22α, by Abcam (1 mentions) Anti 53bp1, by Cell Signaling Technology (1 mentions) Rabbit polyclonal igg, by Abcam (1 mentions) Hematoxylin, by Merck Group (1 mentions) Mounting media, by Agilent Technologies (1 mentions) Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions) Wheat germ agglutinin (wga), by Thermo Fisher Scientific (1 mentions) Metamorph nx software, by Molecular Devices (1 mentions) Ez chamber slides, by Merck Group (1 mentions) Microplate reader, by Molecular Devices (1 mentions) Sm22α, by Agilent Technologies (1 mentions)

19 protocols using microsystems microscope

1

Atherogenesis Induction by PCSK9 Overexpression

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To induce atherogenesis by hypercholesterolemia, 8-10-week-old male Rac1fl/fl or Rac1fl/fl/LC mice were infected with adenoviral vector overexpressing proprotein convertase subtilisin/kexin type 9 (AdPCSK9) at a dose of 2x1011 viral particles/mouse [19 (link)]. These mice were then fed a high-fat diet for 20 weeks and whole aortas were then fixed, pinned and stained with Sudan IV [20 (link)]. Paraffin-embedded mouse aortic arches were sectioned and stained using immunofluorescence with antibodies against CD68 (Abcam) and SM22α (Abcam) and secondary antibodies rat (Jackson ImmunoResearch), and Alexa 647 anti-goat (Jackson ImmunoResearch) [13 (link)]. Immunofluorescence staining intensity was quantified in the entire wall of aortic arches as reported elsewhere [12 (link)]. Pinned whole aortas and aortic arch histological images were captured with a Leica Microsystems microscope and plaques with lipid content was quantified using Biopix software (GU Ventures) [10 (link)]. In Hematoxylin & Eosin (H&E) stained cross sections of aortic arches, intima/media ratios were quantified as described earlier [12 (link)]. Sirius red stain was performed as described earlier [19 (link)]. The percentage of Sirius red positivity and necrotic core areas within intimal thickening formed in Rac1fl/fl and Rac1fl/fl/LC aortas were quantified using Biopix software (GU Ventures).
Bandaru S., Ala C., Ekstrand M., Akula M.K., Pedrelli M., Liu X., Bergström G., Håversen L., Borén J., Bergo M.O, & Akyürek L.M. (2020). Lack of RAC1 in macrophages protects against atherosclerosis. PLoS ONE, 15(9), e0239284.
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2

Quantifying DNA Damage Foci in Cells

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Cells were plated on coverslips that were placed in 35-mm culture dishes. At specified time points after exposure to 4 Gy radiation, cells were fixed in 4% paraformaldehyde for 10 minutes at ambient temperature, briefly washed in phosphate-buffered saline (Mediatech), and placed in 70% ethanol overnight at 4ºC. Fixed cells were permeabilized with 0.1% Igepal for 20 minutes at ambient temperature, blocked in 2% bovine serum albumin (Sigma) for 1 h, and then incubated in anti-53BP1 (1:200, Cell Signaling) or anti-BRCA1 primary antibody (1:500, Santa Cruz) overnight at 4ºC. Cells were then washed four times with phosphate-buffered saline and then incubated for 1 hour in secondary anti-rabbit antibody conjugated to Cy3 (Jackson ImmunoResearch) for 53BP1 foci or in secondary anti-mouse antibody conjugated to FITC or Cy3 (Jackson ImmunoResearch) to visualize immunoreactivity. DNA was stained with 4′,6-diamidino-2-phenylindole (1:1000, Sigma). Immunoreactions were visualized with a Leica Microsystems microscope (Wetzlar, Germany), and foci were counted manually by using ImageJ software (rsbweb.nih.gov/ij/).
Wang L., Zhang P., Molkentine D.P., Chen C., Molkentine J.M., Piao H., Raju U., Zhang J., Valdecanas D.R., Tailor R.C., Thames H.D., Buchholz T.A., Chen J., Ma L., Mason K.A., Ang K.K., Meyn R.E, & Skinner H.D. (2016). TRIP12 as a mediator of human papillomavirus/p16-related radiation enhancement effects. Oncogene, 36(6), 820-828.
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3

Visualizing Mitochondrial Dynamics in C6 Cells

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C6 cells were grown on coverslips in 24-well plates. After treatment with XN for 48 h, cells were incubated with 150 nM Mito-Tracker green and 10 μM Hoechst 33342 for 30 min at 37 °C. The fluorescence signal was visualized by a Leica Microsystems microscope.
Hou S., Song Y., Sun D., Zhu S, & Wang Z. (2021). Xanthohumol-Induced Rat Glioma C6 Cells Death by Triggering Mitochondrial Stress. International Journal of Molecular Sciences, 22(9), 4506.
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4

Tube Formation Assay with HUVEC

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MDA-MB-231 cells were treated with 5 µm CCX771 and/or 100 ng/ml CXCL12, and the media were collected 24 h following treatment. The wells of 96-well plates were pre-coated with Matrigel for 1 h at 37°C. A total of 2×104 HUVECs were seeded onto wells with the aforementioned media for 24 h. Images were captured with a Leica Microsystems microscope (10X objective; Leica Microsystems, Inc., Buffalo Grove, IL, USA). Only perfectly continuous tubes between two branching points were considered as a tube formation. At least 4 fields were tested per well and experiments were repeated three times.
Qian T., Liu Y., Dong Y., Zhang L., Dong Y., Sun Y, & Sun D. (2017). CXCR7 regulates breast tumor metastasis and angiogenesis in vivo and in vitro. Molecular Medicine Reports, 17(3), 3633-3639.
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5

Histomorphometric Analysis of Bone Tissue

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Paraffin-embedded femurs were sectioned (5 μm), deparaffinized, and then stained with hematoxylin and eosin (H&E) for morphological analysis. For immunohistochemistry staining, deparaffinized and dehydrated tissue sections were re-hydrated before being subjected to antigen retrieval, and then blocked with diluted normal serum for at least 1–1.5 h at 25 °C to eliminate nonspecific binding. The slides were incubated with primary antibodies against osteocalcin (OCN) (1:300; Abcam) overnight at 4 °C. After careful washing, the sections were incubated with horseradish peroxidase conjugates to detect positive signals, followed by counterstaining with hematoxylin (Sigma-Aldrich). Slides incubated with polyclonal rabbit IgG (Abcam) served as negative controls. Pictures were captured and monitored using a Leica Microsystems microscope (Leica Microsystems Ltd.), while the ImageJ software (National Institutes of Health) was used to analyze the number or area of adipocytes and osteoblasts.
Chen G., Zhuo Y., Tao B., Liu Q., Shang W., Li Y., Wang Y., Li Y., Zhang L., Fang Y., Zhang X., Fang Z, & Yu Y. (2020). Moderate SMFs attenuate bone loss in mice by promoting directional osteogenic differentiation of BMSCs. Stem Cell Research & Therapy, 11, 487.
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6

Immunofluorescence Assay for DNA Damage Response

Cited 1 time
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Immunofluorescence was measured as described previously (11 (link)). In brief, cells were cultivated on cover slips placed in 35-mm cell culture dishes. At specified time points after exposure to radiation (2 Gy), cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, briefly washed in PBS (Media-tech), and placed in 70% ethanol overnight at 4°C. Then, fixed cells were permeabilized with 0.1% IGEPAL (octylphenoxypolyethoxyethanol) for 20 minutes at room temperature, blocked in 2% BSA (Sigma) for 60 minutes, and then incubated in anti-γH2AX (1:400) or anti-BRCA1 primary antibody (1:400) overnight at 4°C. To assess H2Aub level, cells were permeabilized with 0.4% Triton X-100 for 5 minutes at room temperature, blocked in 5% normal goat serum for 60 minutes, and then incubated in anti-H2Aub (1:1,000) overnight at 4°C. Cells were then washed four times with PBS and incubated for 1 hour in secondary anti-rabbit antibody conjugated with Cy3 (Jackson ImmunoResearch) to visualize γH2AX foci and H2Aub or in secondary anti-mouse antibody conjugated to Cy3 to visualize BRCA1. DNA was stained with DAPI (4′,6-diamidino-2-phenylindole; Sigma) followed by mounting on labeled slides with mounting media (DAKO). Immunoreaction results were visualized with a Leica Microsystems microscope, and foci were counted with ImageJ software (https://imagej.nih.gov/ij/).
Liu X., Kumar M., Yang L., Molkentine D.P., Valdecanas D., Yu S., Meyn R.E., Heymach J.V, & Skinner H.D. (2017). BAP1 Is a Novel Target in HPV-Negative Head and Neck Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(3), 600-607.
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7

Cardiac Morphology and Fibrosis Analysis

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Rat hearts were harvested at indicated time points. LV mass-to-body weight ratio (LVm/BW) was then calculated. Rat hearts and small intestine were fixed in 4% paraformaldehyde, embedded in paraffin, cut into 6 μm sections and stained with hematoxylin and eosin (H&E) to analyze the organizational morphology. Heart samples were stained with either Masson's Trichome or Picrosirius Red to analyze the myocardial fibrosis. To measure myocyte cross-sectional area, cardiac sections were stained with wheat germ agglutinin (WGA, Thermo Fisher Scientific Inc., USA) solution for 10 min, and washed with PBS three times. After that, sections were imaged using a Leica Microsystems microscope and quantified using Image Pro Plus 6.0.
Li Z., Wu Z., Yan J., Liu H., Liu Q., Deng Y., Ou C, & Chen M. (2019). Gut microbe-derived metabolite trimethylamine N-oxide induces cardiac hypertrophy and fibrosis. Laboratory investigation; a journal of technical methods and pathology, 99(3).
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8

Wound Healing Assay for Cell Migration

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Wound-heal assay was performed to monitor the migratory phenotype of PC cells post exposure to drug(s) [46 (link)]. In brief, cells were cultured in 6-well plates (1 × 106 cells per well) until a confluent monolayer was formed. A 200 µl pipette tip was used to scratch the monolayer. The wells were then washed with PBS and images (10× magnification) were captured of the wound at 0 time point with a Leica Microsystems microscope (Buffalo Grove, IL, USA). Images of the wound were then captured at 72 h post exposure to the drug(s). The cell migration (wound closure) was measured by calculating the distance between 4–5 random points within the wound edges.
Khurana N., Chandra P.K., Kim H., Abdel-Mageed A.B., Mondal D, & Sikka S.C. (2020). Bardoxolone-Methyl (CDDO-Me) Suppresses Androgen Receptor and Its Splice-Variant AR-V7 and Enhances Efficacy of Enzalutamide in Prostate Cancer Cells. Antioxidants, 9(1), 68.
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9

Wound Healing Assay for Cancer Cell Migration

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Wound-healing assays were carried out to measure the effect of drugs on the migratory phenotype of PCa cells, as previously described (49 (link)). Briefly, cells were seeded in 6-well plates (1×106 cells per well) and grown until they formed a confluent monolayer. The monolayers were scratched using a 200 µl pipette tip, wells were washed with PBS and images of the wound (0-time point) were captured using a Leica Microsystems microscope (Buffalo Grove, IL, USA). Growth media (CS-FBS) was returned to each culture and treatments were initiated. Change in wound width was captured after 48 h and cell migration (wound closure) was calculated by measuring the distance between 4–5 random points within the wound edges.
Khurana N., Kim H., Chandra P.K., Talwar S., Sharma P., Abdel-Mageed A.B., Sikka S.C, & Mondal D. (2017). Multimodal actions of the phytochemical sulforaphane suppress both AR and AR-V7 in 22Rv1 cells: Advocating a potent pharmaceutical combination against castration-resistant prostate cancer. Oncology Reports, 38(5), 2774-2786.
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10

Quantifying Ileal Histomorphology in Rats

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The ileum from each rat, located 1 cm from the cecum, was carefully harvested and fixed with 4% paraformaldehyde for 24 hours. These tissue samples were then embedded in paraffin and processed to generate 5 µm-thick sections. These sections were subsequently dewaxed and stained with hematoxylin and eosin using standard protocols to analyze the organizational morphology. After staining, sections were inspected under a Leica Microsystems microscope, and measurements were quantified using Image Pro Plus 6.0 software. Crypt depth was measured from the bottom of the crypt to the crypt-villus junction, and villus length was gauged from the crypt-villus junction to the tip of the villus.
Hao Q.Y., Yan J., Wei J.T., Zeng Y.H., Feng L.Y., Que D.D., Li S.C., Guo J.B., Fan Y., Ding Y.F., Zhang X.L., Yang P.Z., Gao J.W, & Li Z.H. (2024). Prevotella copri promotes vascular calcification via lipopolysaccharide through activation of NF-κB signaling pathway. Gut Microbes, 16(1), 2351532.
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