Fluorescence microscope

Manufactured by Olympus

Sourced in Japan, United States, Germany, China, Italy, United Kingdom, Denmark, Switzerland, France

The Olympus Fluorescence Microscope is an optical microscope that uses fluorescence to visualize and analyze samples. It illuminates the specimen with light of a specific wavelength, causing fluorescent molecules within the sample to emit light at a different wavelength, which is then detected and displayed.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Beyotime (154 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (128 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (116 mentions) In situ cell death detection kit, by Roche (93 mentions) Triton x 100, by Merck Group (78 mentions) Ribo lncrna fish probe mix red, by RiboBio (50 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (47 mentions) Paraformaldehyde (pfa), by Merck Group (45 mentions) Bovine serum albumin (bsa), by Merck Group (42 mentions) Dcfh da, by Beyotime (36 mentions) Image pro plus 6, by Media Cybernetics (34 mentions) Alexa fluor 488, by Thermo Fisher Scientific (32 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (31 mentions) Triton x 100, by Beyotime (31 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (27 mentions) Hoechst 33342, by Thermo Fisher Scientific (26 mentions) Hoechst 33342, by Beyotime (25 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (24 mentions) Ros assay kit, by Beyotime (21 mentions) Hoechst 33342, by Merck Group (21 mentions)

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5 377 protocols using fluorescence microscope

Immunofluorescence Protocol for NF-κB, TGF-β1, and Smad3

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For NF-κB (p65) translocation, cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 10 min and then with 5% BSA in PBS for 30 min. Next, Cells were incubated with anti-NF-κB (p65) antibody (1:200) overnight at 4 °C. Goat Anti-Rabbit IgG H&L (FITC) (1:1000) was used for detection. Immunofluorescence was viewed and captured using Olympus fluorescence microscope.
For TGF-β1 staining, cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 10 min and then with 5% BSA in PBS for 30 min. Next, Cells were incubated with anti-TGF-β1antibody (1:200) overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (TRITC) (1:1000) was used for detection. Immunofluorescence was viewed and captured using Olympus fluorescence microscope.
For Smad3 translocation, cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 10 min and then with 5% BSA in PBS for 30 min. Next, Cells were incubated with anti-Smad3 antibody (1:200) overnight at 4 ℃. Goat Anti-mouse IgG H&L (FITC) (1:1000) was used for detection. Immunofluorescence was viewed and captured using Olympus fluorescence microscope.

Ye S., Huang H., Xiao Y., Han X., Shi F., Luo W., Chen J., Ye Y., Zhao X., Huang W., Wang Y., Lai D., Liang G, & Fu G. (2023). Macrophage Dectin-1 mediates Ang II renal injury through neutrophil migration and TGF-β1 secretion. Cellular and Molecular Life Sciences, 80(7), 184.

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Quantifying Oxidative Stress in Aortic Cells

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Fresh frozen sections of aortic root were incubated with 10 µM DHE (Thermo Fisher Scientific) in a humidified chamber protected from light for 0.5 h at 37 °C, and then photographed under a fluorescence microscope (Olympus Optical). Meanwhile, the adjacent sections of serial sections were stained for HE. hVSMCs were incubated with 10 µM DCFH‐DA (Sigma‐Aldrich) in a humidified chamber protected from light for 0.5 h at 37 °C, and then immediately photographed under a fluorescence microscope (Olympus Optical).

Bi X., Du C., Wang X., Wang X., Han W., Wang Y., Qiao Y., Zhu Y., Ran L., Liu Y., Xiong J., Huang Y., Liu M., Liu C., Zeng C., Wang J., Yang K, & Zhao J. (2021). Mitochondrial Damage‐Induced Innate Immune Activation in Vascular Smooth Muscle Cells Promotes Chronic Kidney Disease‐Associated Plaque Vulnerability. Advanced Science, 8(5), 2002738.

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Quantification of Cellular Markers in RSC96 and L929 Cells

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RSC96 cells were fixed with 4% paraformaldehyde, permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100, blocked with PBS containing 0.1% Tween and 5% goat serum, and then stained with rabbit anti-rat p75 (1:100, Abcam), rabbit anti-rat Ki67 (1:100, Abcam), and rabbit anti-rat c-Jun (1:100, Abcam) primary antibodies. The coverslips were then sequentially labeled with species-specific fluorochrome-conjugated secondary antibodies and DAPI (Sigma-Aldrich, St. Louis, MO, United States). Samples were visualized using a fluorescence microscope (Olympus). The numbers of p75⁺ cells, Ki67⁺ cells, and c-Jun⁺ cells were counted at ×400 original magnification.
L929 cells were fixed with 4% paraformaldehyde, permeabilized with PBS containing 0.1% Triton X-100, blocked with PBS containing 0.1% Tween and 5% goat serum, and then stained with rabbit anti-mouse α-SMA (1:100, Abcam) and rabbit anti-mouse Ki67 (1:100, Abcam) primary antibodies. The coverslips were then sequentially labeled with species-specific fluorochrome-conjugated secondary antibodies and DAPI (Sigma-Aldrich). Samples were visualized using a fluorescence microscope (Olympus). The numbers of α-SMA⁺ and Ki67⁺ cells were counted at ×400 original magnification.

Zhou S., Wan L., Liu X., Hu D., Lu F., Chen X, & Liang F. (2022). Diminished schwann cell repair responses play a role in delayed diabetes-associated wound healing. Frontiers in Physiology, 13, 814754.

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Transgenic Silkworm Screening and Analysis

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The transgenic silkworms were screened using a fluorescence microscope (Olympus, Tokyo, Japan), and the genome was extracted as a template. PCR amplification was performed with BmEcKL1 target site detection primers (Table S1). The PCR fragment was subjected to agarose gel electrophoresis, purified, connected to T vector, transformed in E. coli, and single clones were sequenced by Sanger method using M13F/R primer. WT and transgenic silkworms were reared to the fifth instar for dissection, after which the silk glands were excised, washed with phosphate-buffered saline, and imaged using a fluorescence microscope (Olympus, Tokyo, Japan). The WT and transgenic silkworms were mixed and reared to the fifth instar, and their body weights were counted. The silkworms were reared to the moth stage, and cocoons were cut in advance to identify males and females.

Li S., Lao J., Sun Y., Hua X., Lin P., Wang F., Shen G., Zhao P, & Xia Q. (2024). CRISPR/Cas9-Mediated Editing of BmEcKL1 Gene Sequence Affected Silk Gland Development of Silkworms (Bombyx mori). International Journal of Molecular Sciences, 25(3), 1907.

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Immunofluorescence Analysis of Lung Tissue and Cell Cultures

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The lung tissues were repaired with citric acid antigen and washed with PBS and then incubated with the antibodies of SDC-1, TGFβ1, p-Smad3, collagen I, and α-SMA at 4°C for 15 h. The lung tissues were washed with PBS and then stained with Alexa Fluor goat anti-rabbit and DAPI. Finally, the lung tissues were observed and photographed under a fluorescence microscope (Olympus, Japan).
The Beas-2B or HLF-1 cells were seeded on soul plates in 24-well plates. The cells were fixed with 4% paraformaldehyde after different pretreatments and treatments and incubated with the antibody of SDC-1 overnight at 4°C. The soul plates were washed with PBS and then stained with Alexa Fluor goat anti-rabbit and DAPI. Finally, the cell images were photographed under a fluorescence microscope (Olympus, Japan).

Zhang D., Qiao X.R., Cui W.J., Zhang J.T., Pan Y., Liu X.F, & Dong L. (2021). Syndecan-1 Amplifies Ovalbumin-Induced Airway Remodeling by Strengthening TGFβ1/Smad3 Action. Frontiers in Immunology, 12, 744477.

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Mitochondrial Membrane Potential and Mass Analysis

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Mitochondrial membrane potential in collected EA.hy926 cells was examined by fluorescent dye JC-1. JC-1 is a fluorescent lipophilic carbocyanine dye used to measure mitochondrial membrane potential. When the mitochondrial membrane potential is high, JC-1 aggregates in the matrix to form J-aggregates, which can produce red fluorescence. When the mitochondrial membrane potential is low, JC-1 cannot aggregate in the mitochondrial matrix, the presence of monomers produces green fluorescence. In short, the cells were stained with JC-1 for 30 min at 37 °C, and the green JC monomers (488 nm) and red JC aggregates (570 nm) were visualized by a fluorescence microscope (Olympus, Japan). To assess the mitochondrial mass in ECs, the collected EA.hy926 cells were incubated with NAO (0.1 μM) for 30 min at 37 °C under a dark environment, and the cellular fluorescence was imaged using a fluorescence microscope (Olympus, Japan).

Wang Z.C., Niu K.M., Wu Y.J., Du K.R., Qi L.W., Zhou Y.B, & Sun H.J. (2022). A dual Keap1 and p47phox inhibitor Ginsenoside Rb1 ameliorates high glucose/ox-LDL-induced endothelial cell injury and atherosclerosis. Cell Death & Disease, 13(9), 824.

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Chromatin Condensation Visualization

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To visualize chromatin condensation, we used Hoechst33342 or DAPI to stain DNA in the nuclei. Briefly, PC3 cells cultured on cover glasses were incubated with 5 μg/mL Hoechst33342 or DAPI for 15 minutes. The cells were then washed with PBS and nuclear fluorescence was detected using fluorescence microscope (Olympus). Alternatively, apoptotic cells were identified using an in situ cell death detection TUNEL kit (Roche). The staining was performed according to manufacturer's instruction and observed using fluorescence microscope (Olympus).

Wang Y., Niu H., Hu Z., Zhu M., Wang L., Han L., Qian L., Tian K., Yuan H, & Lou H. (2018). Targeting the lysosome by an aminomethylated Riccardin D triggers DNA damage through cathepsin B‐mediated degradation of BRCA1. Journal of Cellular and Molecular Medicine, 23(3), 1798-1812.

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Cytoskeleton and Proliferation of BMSCs

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The BMSCs were implanted on plates and held in full culture medium for 1, 3, and 7 days. The cytoskeleton morphology was evaluated, where the cells were washed with cold 1X PBS for 2–3 min, fixed in 4% paraformaldehyde (PFA) at room temperature (RT) for 15 min and permeabilized with phosphate buffered saline containing 0.1% Triton X (PBS-Tx-0.1%). The cytoskeleton was stained with tetramethylrhodamine isothiocyanate (TRITC)-phalloidin (Yeasen, China), while the nuclei were stained with 4′,6-Diamidino-2-phenylindol (DAPI, ThermoFisher Scientific, USA). The samples were imaged using a fluorescence microscope (Olympus, Japan). The BMSC proliferation induced by different pore channels was evaluated, where cells were cultured in EdU-containing medium (diluted to 1: 1000, 1 mg L-1; Ribobio, China). The samples were collected, and the plates were stained using a 5-ethynyl-2′-deoxyuridine (EdU) Apollo kit (Ribobio, China) according to the manufacturer's instructions, while the cell nuclei were counterstained with Hoechst 33342. The samples were observed using a fluorescence microscope (Olympus, Japan), where statistical analysis of the images was used to calculate the percentage of EdU positive cells.

Yin S., Zhang W., Tang Y., Yang G., Wu X., Lin S., Liu X., Cao H, & Jiang X. (2020). Preservation of alveolar ridge height through mechanical memory: A novel dental implant design. Bioactive Materials, 6(1), 75-83.

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Subcutaneous Tumor Xenograft Analysis

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The subcutaneous xenografts were surgically excised from BALB/c nude mice, paraformaldehyde fixed and paraffin embedded, and sectioned. After dewaxing, rehydration, and antigen retrieval, the slides were blocked in 5% BSA for 40 min and incubated with primary antibodies at 4 °C overnight: Fibrin(ogen) (Novus, USA), DAB2IP (Proteintech, USA), Nanog (Proteintech, USA). Slides were then incubated with HRP-conjugated or CY3-conjugated anti-rabbit secondary antibody (Servicebio, China) for immunohistochemistry (IHC) or immunofluorescence (IF) assays. DAPI was used to stain the nucleus.
IHC images were acquired under white light of fluorescence microscope (Olympus, Japan) and Image-pro plus 6.0 software was used to quantify the corresponding staining. IF staining was recorded by fluorescence microscope (Olympus, Japan).

Zhang M., Xu C., Wang H.Z., Peng Y.N., Li H.O., Zhou Y.J., Liu S., Wang F., Liu L., Chang Y., Zhao Q, & Liu J. (2019). Soft fibrin matrix downregulates DAB2IP to promote Nanog-dependent growth of colon tumor-repopulating cells. Cell Death & Disease, 10(3), 151.

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Immunofluorescent Retinal Cell Imaging

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Eyeballs were embedded in OCT and sectioned by Leica cryostat. After blocking, sections were incubated with anti-Nox4 primary antibody (Santa-Cruz Biotechnology Inc., Santa Cruz, CA), anti-CD31 (BD Pharmingen, San Diego, CA), or anti-p-ERK (Cell Signaling Technology, Boston, MA) overnight. Signals were detected with Alexa Fluor 488 or Alexa Fluor 594 conjugated secondary antibodies (Invitrogen, Carlsbad, CA). Slides were mounted with VECTASHIELD mounting medium with DAPI (Vector Laboratories, Burlingame, CA) and visualized under fluorescence microscope (Olympus). Sections with omission of primary antibodies were used as negative controls. For retinal flat-mount staining, retinas were dissected out and permeabilized followed by incubation with anti-Nox4 antibody (Santa-Cruz) and anti-CD31 (BD Pharmingen). After intensive rinse, retinas were stained with Alexa Fluor 488 and Alexa Fluor 594 conjugated secondary antibody (Invitrogen). The retinas were flat-mounted and observed under fluorescence microscope (Olympus).

Li J., Wang J.J, & Zhang S.X. (2015). NADPH Oxidase 4-Derived H2O2 Promotes Aberrant Retinal Neovascularization via Activation of VEGF Receptor 2 Pathway in Oxygen-Induced Retinopathy. Journal of Diabetes Research, 2015, 963289.

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