Dm500 fluorescence microscope

Manufactured by Leica

Sourced in Germany

The Leica DM500 is a fluorescence microscope designed for routine imaging and analysis. It features a LED illumination system and a range of optical components to enable fluorescence observation and imaging.

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Lab products found in correlation

Gemini em microplate reader, by Molecular Devices (1 mentions) 96 well plate, by Thermo Fisher Scientific (1 mentions) Dmem medium, by Merck Group (1 mentions) Ab150157, by Abcam (1 mentions) Ab150080, by Abcam (1 mentions) Nf κb p65 antibody, by Cell Signaling Technology (1 mentions) Ab104139, by Abcam (1 mentions) Ab6640, by Abcam (1 mentions) Ab150077, by Abcam (1 mentions) Oil red o, by Beyotime (1 mentions) Oil red o, by Leica (1 mentions)

Close spelling variants of this product

Fluorescence microscope (2465 citations) DM500 (342 citations) DM500 microscope (102 citations) DM500 B Fluorescence microscope (8 citations) DM500 B fluorescence confocal microscope (2 citations)

6 protocols using dm500 fluorescence microscope

Microscopic Examination of Cells

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Phase-contrast and fluorescence microscopy examinations were conducted with a LEICA DM500 fluorescence microscope. Cell culture medium was vacuumed from the wells in the 12-well plate and replaced with a 3.7% formalin solution. After one minute, the formalin solution was removed and replaced with a 100% ice-cold methanol solution. After one minute, the methanol solution was removed from each well and replaced with Phosphate-buffered (PBS) solution. After 15 minutes, the PBS solution was removed from each well and replaced with DAPI nuclear DNA staining solution.
The cells in each well then underwent microscopic observation. Phase-contrast microscopy was applied to examine cell morphology. Fluorescence microscopy was used to examine nuclear morphology (DAPI dye). In the case of dye-doped silica NSs, because they naturally fluoresce, a fluorescence microscope was used to determine if the Nanoparticles (NPs) were incorporated into the cells.

Larner S.F., Wang J., Goodman J., O’Donoghue Altman M.B., Xin M, & Wang K.K. (2017). In Vitro Neurotoxicity Resulting from Exposure of Cultured Neural Cells to Several Types of Nanoparticles. Journal of Cell Death, 10, 1179670717694523.

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Measuring NADPH Oxidase-Derived Superoxide

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16HBE cells were treated with 500 μM of PQ for 0 h, 12 h, 24 h, and 48 h; L-012 dye was used to detect extracellular NADPH oxidase-derived superoxide. In brief, 16HBE cells were diluted into approximately 4–6 × 10⁴ cells/well into 96-well plates (Thermo, USA) in phenol-free DMEM medium (Sigma, USA) with L-012 at the concentration of 500 μM according to our preliminary experiments (data not shown) for 10 min and luminescence was detected by a Gemini EM microplate reader (Molecular Devices, USA) at the excitation wavelength of 488 nm and emission wavelength of 525 nm respectively. Cellular ROS levels were next measured by dihydroethidium (DHE) staining. Cells were washed with PBS twice and diluted; 10 μM of DHE (Invitrogen, USA) was selected according to our preliminary experiments (data not shown) to incubate with the cells for 30 min at 37 °C without light exposure. After incubation, cells were washed with PBS and DM500 fluorescence microscope (Leica, Germany) was employed to observe ROS productions. The fluorescence intensity was quantified and calculated by ImageJ software.

Yao J., Zhang J., Tai W., Deng S., Li T., Wu W., Pu L., Fan D., Lei W., Zhang T, & Dong Z. (2019). High-Dose Paraquat Induces Human Bronchial 16HBE Cell Death and Aggravates Acute Lung Intoxication in Mice by Regulating Keap1/p65/Nrf2 Signal Pathway. Inflammation, 42(2), 471-484.

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Intracellular R6G Accumulation Assay

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R6G intracellular accumulation was assessed as previously mentioned with some modifications [34 (link)]. Briefly, the R6G-loaded cells were treated with FA (1000 μg/mL) followed by glucose addition (10 mM final concentration) and incubation for 20 min as previously mentioned. The cells were then collected by centrifugation at 5000 × g for 5 min at 4 °C before being rinsed twice with cold PBS. The resulting pellet was examined using a Leica DM500 fluorescence microscope, Leica, Germany. The Image J software was used to measure the fluorescence intensity.

Gomaa S.E., Abbas H.A., Mohamed F.A., Ali M.A., Ibrahim T.M., Abdel Halim A.S., Alghamdi M.A., Mansour B., Chaudhary A.A., Elkelish A., Boufahja F., Hegazy W.A, & Yehia F.A. (2024). The anti-staphylococcal fusidic acid as an efflux pump inhibitor combined with fluconazole against vaginal candidiasis in mouse model. BMC Microbiology, 24, 54.

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Immunohistochemical Analysis of ETV6 and F4/80 in Aorta

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Aortas were fixed in 4% formaldehyde for 24 h, dehydrated overnight, and embedded in paraffin. Paraffin cross-sections (4-μm thick) were prepared. After incubation with 5% normal goat serum for 1 h at room temperature, the sections were incubated with rabbit anti-ETV6 antibody (1:200, PA5-109,697, Thermo Fisher Scientific) and rat anti-F4/80 (10 μg/ml, ab6640, Abcam) overnight at 4^oC. After three washes with PBS, the samples were incubated with 5 μg/ml Alexa Fluor® 488-conjugated goat anti-rat IgG H&L and Alexa Fluor® 594-conjugated goat anti-rabbit IgG H&L (1:1000, ab150157, ab150080, Abcam) for an hour at room temperature. For Immunofluorescence cytochemistry, macrophages were fixed in acetone for 10 min and incubation with 5% normal goat serum for 1 h at room temperature. Macrophages were then incubated with NF-κB p65 antibody (1:500, #8242, Cell Signaling Technology) overnight at 4^oC. After three washes with PBS, cells were incubated with Alexa Fluor® 488-conjugated goat anti-rabbit IgG H&L (1:1000, ab150077, Abcam) for an hour at room temperature. Cells were washed and incubated in the mounting medium with DAPI (ab104139, Abcam). The samples were observed and photographed on a Leica DM500 fluorescence microscope (Leica).

Xiong X., Yan Z., Jiang W, & Jiang X. (2022). ETS variant transcription factor 6 enhances oxidized low-density lipoprotein-induced inflammatory response in atherosclerotic macrophages via activating NF-κB signaling. International Journal of Immunopathology and Pharmacology, 36, 20587384221076472.

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Histological Analysis of Mouse Liver

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Dissected liver samples from each mouse were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned at 5 μm, and stained with hematoxylin and eosin (H&E) after deparaffinization. H&E-stained sections were imaged under Leica DM500 fluorescence microscope (Leica, Buffalo Grove, IL) at 20X magnification.

Le H.H., Lee M.T., Besler K.R, & Johnson E.L. (2022). Host hepatic metabolism is modulated by gut microbiota–derived sphingolipids. Cell host & microbe, 30(6), 798-808.e7.

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Quantifying Aortic Atherosclerosis in Mice

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The mice were euthanized by CO₂ inhalation. The aortas were taken, fixed in 4%
formaldehyde, and then immersed in 5% oil red O (Beyotime Biotech) for half an hour. After
three washes in deionized water, atherosclerotic lesions, that is, oil red O-positive
areas along the aorta, were recorded on a Leica DM500 fluorescence microscope (Leica).

Lin G., Zhang L., Yan Z., Jiang W., Wu B., Li D, & Xiong X. (2022). Identification of heterogeneous subsets of aortic interleukin-17A-expressing CD4+ T cells in atherosclerotic mice. International Journal of Immunopathology and Pharmacology, 36, 03946320221117933.

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