Lcx100 imaging system

Manufactured by Olympus

Sourced in Japan

The LCX100 Imaging System is a laboratory equipment designed for high-quality imaging and analysis. It features a compact and integrated design, allowing for efficient and convenient use in various research and clinical settings.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Dab substrate, by ZSGB-BIO (2 mentions) Lucigenin, by Merck Group (1 mentions) Fb12 berthold luminometer, by Berthold Technologies (1 mentions) Image pro plus software, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Ros assay kit, by Beyotime (1 mentions) Total superoxide dismutase assay kit, by Beyotime (1 mentions) Lipid peroxidation mda assay kit, by Merck Group (1 mentions) Alexa 546 labeled anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions) Anti nf κb p65 antibody, by Cell Signaling Technology (1 mentions) Crystal violet, by Santa Cruz Biotechnology (1 mentions) Modified 24 well boyden chambers, by Corning (1 mentions) Dab substrate, by Solarbio (1 mentions) Dcfh da, by Beyotime (1 mentions) Mouse anti cd31, by Abcam (1 mentions) Alexa fluor 488 goat anti rabbit, by Abcam (1 mentions) Maxvision hrp polymer anti mouse rabbit ihc kit, by Maixin Group (1 mentions) Iba 1, by Abcam (1 mentions) Alexa488 conjugated donkey anti goat, by Thermo Fisher Scientific (1 mentions)

17 protocols using lcx100 imaging system

Quantifying Brain Superoxide Levels

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Dihydroethidium (DHE) staining was used for superoxide detection. The whole brains were removed and immediately embedded in Tissue-Tek^® OCT compound, after which they were frozen in an isopentane liquid bath on dry ice for 10 min. The samples were cut into 10-μm-thick sections, placed on glass slides, and incubated in 1 M DHE (Thermo Fisher, dissolved in PBS) for 30 min in a light-protected humidified chamber at room temperature. Slices were imaged using an Olympus LCX100 Imaging System and analyzed with Image J software.
In addition, lucigenin chemiluminescence was applied as an indicator of brain superoxide production. PVN samples were isolated from the brains and homogenized in PBS. Then, specimens were transferred into a polypropylene tube containing PBS-HEPES buffer and lucigenin (Sigma Aldrich, at a final concentration of 0.2 mM). Measurements were performed with an FB12-Berthold luminometer (Berthold Technologies, Bad Wildbad, Germany) at room temperature. The RLU emitted was recorded and integrated over 30-s intervals for 5 min. Activity was normalized to tissue weights.

Wang Y., Hu H., Yin J., Shi Y., Tan J., Zheng L., Wang C., Li X., Xue M., Liu J., Wang Y., Li Y., Li X., Liu F., Liu Q, & Yan S. (2019). TLR4 participates in sympathetic hyperactivity Post-MI in the PVN by regulating NF-κB pathway and ROS production. Redox Biology, 24, 101186.

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Sympathetic Activation and Fos Expression After Myocardial Infarction

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The expression of central sympathetic proteins was detected by immunohistochemistry. Central sympathetic activity was measured by Fos family assays after MI. Fos family proteins were quantified using a mouse monoclonal anti‐c‐Fos (E‐8) antibody, which recognizes c‐Fos, Fos‐B, Fra‐1 and Fra‐2 proteins. Tissue slices were probed with the primary antibody c‐Fos (E‐8) mouse monoclonal antibody (1:50, Santa Cruz, CA, USA) overnight at 4℃. Sections were incubated with DAB substrate (ZSGB‐BIO; Beijing, China) and then counterstained with haematoxylin. Images were obtained using an LCX100 imaging system (Olympus). The number of METTL3 and Fos‐positive cells in the bilateral boundary of the PVN was analysed by Image‐Pro Plus 6.0.

Qi L., Hu H., Wang Y., Hu H., Wang K., Li P., Yin J., Shi Y., Wang Y., Zhao Y., Lyu H., Feng M., Xue M., Li X., Li Y, & Yan S. (2022). New insights into the central sympathetic hyperactivity post‐myocardial infarction: Roles of METTL3‐mediated m6A methylation. Journal of Cellular and Molecular Medicine, 26(4), 1264-1280.

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Histological Evaluation of Pulmonary Fibrosis

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At 3, 6, 10, and 20 weeks after radiation, mice selected from the groups were sacrificed and their lung tissues were harvested for histological examination. The lung samples were fixed in 4% paraformaldehyde and embedded in paraffin. The samples were sectioned for staining with hematoxylin and eosin (H&E) or Masson’s trichrome stain to assess the fibrosis score according to standard protocols. The histopathological evaluation of pulmonary fibrosis was scored according to the density of Masson’s staining (Jiancheng, China) using an image J analysis program. Ten microscopic fields were randomly selected for analysis. Assessment was performed in a double-blind fashion. All images were acquired using an Olympus LCX100 Imaging System and analyzed using Imagepro plus software (Olympus Corporation, Tokyo, Japan).

Hu D., Zhang Y., Cao R., Hao Y., Yang X., Tian T, & Zhang J. (2020). The protective effects of granulocyte-macrophage colony-stimulating factor against radiation-induced lung injury. Translational Lung Cancer Research, 9(6), 2440-2459.

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Histological and Immunohistochemical Analysis of Tumor Tissues

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For histology, tumor tissues were fixed in 4% formalin, embedded in paraffin, sliced into 4-µm sections and stained with hematoxylin and eosin (H&E).
For IHC staining, after a set of processes including dewaxing, rehydration, antigen retrieval with citrate buffer pH 6.0, endogenous peroxidase blocking with 3% H₂O₂ for 10 minutes, followed by incubating with a primary anti-C5 antibody (Affinity, Amherst, NH, USA; 1:50) overnight at 4 ℃ and then with a secondary antibody (Zhongshan Goldenbridge Biotechnology Company, Beijing, China; catalog No. SP-9001) for 30 minutes at 37 ℃. Immunodetection of the slides was performed by 3,3'-diaminobenzidine (DAB). All images were captured with an Olympus LCX100 Imaging System (Olympus, Tokyo, Japan). Results were analyzed with ImageJ software (version 1.62; NIH).

Yuan M., Wang C., Wu Y., Qiao L., Deng G., Liang N., Chen F., Liu L., Chen Y., Yang Y., Wang H., Liu T., Yang X., Zhang Y., Lv Y., Suwinski R., Hu P., Zhang Y, & Zhang J. (2023). Targeting complement C5a to improve radiotherapy sensitivity in non-small cell lung cancer. Translational Lung Cancer Research, 12(5), 1093-1107.

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Quantifying Oxidative Stress in BV-2 Cells

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The ROS level of BV-2 cells (1×10⁶/well) was detected by ROS assay kit (Beyotime Biotechnology, Shanghai, China) according to the manufacturer’s instructions. Briefly, BV-2 cells were cultured in DMEM containing 20 μM Aβ and/or 20 μMMeJA for 48 hr. Then, the medium was replaced by serum-free DMEM containing 10 μM DCFH-DA probe and after that, the BV-2 cells were cultured for another 30 min in darkness. The cells were washed with 1×PBS and stained with 1 μg/mL DAPI (Sigma-Aldrich, USA) for 2 min. Then, the cells were photographed by LCX100 Imaging System (Olympus, Japan) with wavelength of 488 nm. Parallel experiments were performed in triplicate for each group.

Li H., Lv L., Wu C., Qi J, & Shi B. (2020). Methyl Jasmonate Protects Microglial Cells Against β-Amyloid-Induced Oxidative Stress and Inflammation via Nrf2-Dependent HO-1 Pathway. Neuropsychiatric Disease and Treatment, 16, 1399-1410.

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Evaluation of Oxidative Stress Markers in Aβ-Treated BV-2 Cells

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According to the information in cell culture and treatment section, four groups of BV-2 cells (control, Aβ, MeJA, and Aβ+MeJA) were setup. The cells were collected and homogenized when the monolayer was formed. Lipid Peroxidation MDA Assay Kit (Sigma-Aldrich, USA) was used to detect MDA according to the manufacturer’s instructions. Briefly, the related reagents were added into the homogenates in order, followed by incubation at 95°C for 40 min. The mixture was cooled on ice and centrifuged at 4000 rpm for 10 min. 200 μL of supernatant was transferred to 96-well plates and the absorbance was detected at 535 nm. Total Superoxide Dismutase Assay Kit (Beyotime Biotechnology, China) was used to determine the SOD activity according to the manufacturer’s instructions. Briefly, test reagents were added to the homogenate and then incubated at 37°C for 20 min. After centrifugation at 4000 rpm for 10 min, the supernatant was transferred into a 96-well plate and the absorbance was detected at 450 nm by LCX100 Imaging System (Olympus, Japan).

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Investigating NF-κB Activation in HRGECs

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HRGECs were allowed to grow to confluence on fibronectin-coated glass chamber slides and exposed to Cd for 24 h. The monolayers of HRGECs were then washed with PBS containing 100 mM glycine, fixed with 4% paraformaldehyde for 5 min, and washed three times with PBS for 10 min. Immunoreactivity was examined by staining with a rabbit polyclonal anti NF-κB p65 antibody (1:400; Cell Signaling Technology) overnight at 4°C and incubation with an Alexa 546-labeled anti-rabbit secondary antibody (1:200; Molecular Probes, Eugene, OR, USA) for 2 h. Nuclear staining was achieved using DAPI (1:1,000; Molecular Probes) and images of the the samples were captured using an Olympus LCX100 Imaging system (Olympus Corp., Tokyo, Japan) with an excitation wavelength of 546 nm.

Zhang H., Li L., Wang Y., Dong F., Chen X., Liu F., Xu D., Yi F., Kapron C.M, & Liu J. (2016). NF-κB signaling maintains the survival of cadmium-exposed human renal glomerular endothelial cells. International Journal of Molecular Medicine, 38(2), 417-422.

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Cell Migration Assay with VEGF Chemoattractant

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Cell migration assays were performed using modified 24-well Boyden chambers (Costar, Acton, MA, USA), containing a polycarbonate membrane with 8.0-μm pores. HUVECs were starved in basic EGM-2 (serum/growth factor free) overnight at 37°C, prior to being harvested with trypsin and resuspended in basic EGM-2. The single cell suspensions with 20 μM DHA or 20 μM SB203850 were seeded at 1×10⁵ cells/well in the upper chamber, while 0.5 ml EGM-2 with 20 ng/ml VEGF was added to the bottom chamber as chemoattractants. After 24 h incubation, the migrated cells on the bottom surface were stained with 0.1% crystal violet (Santa Cruz Biotechnology, Inc.) and counted under an Olympus LCX100 Imaging system (Olympus Corporation, Tokyo, Japan).

GUO L., DONG F., HOU Y., CAI W., ZHOU X., HUANG A.L., YANG M., ALLEN T.D, & LIU J. (2014). Dihydroartemisinin inhibits vascular endothelial growth factor-induced endothelial cell migration by a p38 mitogen-activated protein kinase-independent pathway. Experimental and Therapeutic Medicine, 8(6), 1707-1712.

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Sympathetic Nerve Activity Post-MI in PVN

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For immunohistochemical studies, brain tissue was cut into 4-μm-thick sections to measure sympathetic nerve activity via Fos family detection post-MI in the PVN. Fos family was measured using a rabbit polyclonal antibody c-fos K-25, which recognizes Fos, Fos-B, Fra-1, and Fra-2 proteins. The sections were incubated with the primary antibody: anti-c-fos K-25 rabbit polyclonal antibody (1:1000, Santa Cruz Biotechnology, San Francisco, CA, USA) overnight at 4 °C, and then HRP conjugated secondary antibodies: goat anti-rabbit IgG for 1 h at room temperature. The slices were incubated with DAB substrate (Solarbio Science & Technology Co., Ltd. Beijing, China) and then counterstained with hematoxylin. The digitized pictures were obtained using an Olympus LCX100 Imaging System. The number of Fos-positive cells was measured.

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Measuring Oxidative Stress in H9c2 Cells

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Following treatment with LPS, the culture medium was removed, and H9c2 cells were subsequently incubated with 1 ml of serum-free DMEM supplemented with 1 µl of fluorescent dichloro-dihydro-fluorescein diacetate (DCFH-DA; Beyotime Institute of Biotechnology) at 37˚C for 30 min in the dark. Cells were washed twice with PBS and fluorescence was observed under an Olympus LCX100 imaging system (Olympus Corporation). The average fluorescence intensity was measured using ImageJ v1.8.0.112 software (National Institutes of Health).

Feng D., Guo L., Liu J., Song Y., Ma X., Hu H., Liu J, & Hao E. (2021). DDX3X deficiency alleviates LPS-induced H9c2 cardiomyocytes pyroptosis by suppressing activation of NLRP3 inflammasome. Experimental and Therapeutic Medicine, 22(6), 1389.

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