Bx41 fluorescence microscope

Manufactured by Olympus

Sourced in Japan, United States

The BX41 is a fluorescence microscope manufactured by Olympus. It is designed for observing and analyzing samples using fluorescence techniques. The BX41 incorporates optical components and illumination systems to enable fluorescence imaging, but a detailed description of its specific features and capabilities is not provided in order to maintain an unbiased and factual approach.

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Lab products found in correlation

Image pro plus analysis software 6, by Media Cybernetics (5 mentions) Triton x 100, by Merck Group (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Methanol, by Thermo Fisher Scientific (4 mentions) In situ cell death detection kit, by Roche (4 mentions) Tween 20, by Merck Group (3 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (2 mentions) Glial fibrillary acidic protein (gfap), by Santa Cruz Biotechnology (2 mentions) Rabbit anti mouse α sma antibody, by Abcam (2 mentions) Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Goat anti at2r receptor polyclonal antibody, by Santa Cruz Biotechnology (2 mentions) Dp71 digital camera, by Olympus (2 mentions) Dp71 camera, by Olympus (2 mentions) Vectashield, by Vector Laboratories (2 mentions) Cy2 or cy5 labeled secondary antibodies, by Jackson ImmunoResearch (2 mentions) A11001, by Thermo Fisher Scientific (2 mentions) Ab8295, by Abcam (2 mentions) Livedead fixable, by Thermo Fisher Scientific (2 mentions) Td 20 20 luminometer, by Turner Designs (2 mentions) F view 2 ccd camera, by Olympus (2 mentions)

Close spelling variants of this product

Fluorescence microscope (5377 citations) BX41 microscope (1002 citations) BX41 and BX51 fluorescence microscopes (5 citations)

102 protocols using bx41 fluorescence microscope

Immunostaining and Quantitative Analysis

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The analysis was performed as described [31 (link)]. Briefly, coverslips containing 100–200 cells/mm² were fixed with (100%) methanol followed by treatment with cold ethanol and two rinses in phosphate-buffered saline (PBS). Samples were blocked with 3%bovine serum albumin (BSA) in PBS/Tween 20 (PBST) for 30 min and incubated in PBST containing 1%BSA, anti-IL-1Ra, and anti-GFAP antibodies. For dilutions of primary antibodies, please see Table 1. After three washes in PBST (15 min each), slides were further incubated with Cy2 or Cy5 (Jackson ImmunoResearch). The samples were mounted and observed under an Olympus BX-41 fluorescence microscope.
Immunostaining of tissue was performed by fixing the brains in 4%paraformaldehyde followed by 30%sucrose [32 (link)]. Tissue was sealed into OCT and sectioned every 40 microns on a Leica Cryostat CM3050 S and kept in cryoprotectant. Frozen sections were treated with cold ethanol (–20°C), followed by two rinses in PBS, blocking with 2%BSA in PBST, and double-labeling with two primary antibodies (Table 1). After three washes in PBST, sections were further incubated with Cy2 and Cy5 (Jackson ImmunoResearcdh Laboratories). The samples were mounted and observed under an Olympus BX-41 fluorescence microscope. Counting analysis was performed using Olympus Microsuite V software with the help of a touch counting module.

Chakrabarti S., Prorok T., Roy A., Patel D., Dasarathi S, & Pahan K. (2021). Upregulation of IL-1 Receptor Antagonist by Aspirin in Glial Cells via Peroxisome Proliferator-Activated Receptor-Alpha. Journal of Alzheimer's Disease Reports, 5(1), 647-661.

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Carbenoxolone Inhibits Hemichannel-Mediated Dye Uptake

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Hearts were rapidly excised and perfused through the aorta with Krebs–Henseleit buffer. After an equilibration period (10-min), hearts were treated with the vehicle or hemichannel blocker carbenoxolone (10 µmol/L, Sigma-Aldrich Chemical, St Louis, MO, USA) for 10 min. After this period, hearts were perfused with ethidium bromide (BrEt) 15 µmol/L (Sigma-Aldrich Chemical, St Louis, MO, USA) for 15 min, in the same medium (with or without blocker) followed by a 15-min wash period. Then, hearts were embedded in 2-methylbutane and frozen in liquid nitrogen. Cryosections (8 μm) were obtained, washed with PBS and fixed with 4% paraformaldehyde. After fixation, the sections were washed with PBS, mounted using Prolong gold antifade reagent containing 4′,6-diamidino-2-phenylindole (DAPI) (Invitrogen, Thermo Fisher Scientific, Carlsbad, CA, USA), observed with a BX41 Olympus fluorescence microscope and photographed with a ProgRes C5 digital camera (Olympus Corporation, Tokyo, Japan). The intensity of the signal was evaluated using the ImageJ program (NIH public domain software).

Vielma A.Z., Boric M.P, & Gonzalez D.R. (2020). Apocynin Treatment Prevents Cardiac Connexin 43 Hemichannels Hyperactivity by Reducing Nitroso-Redox Stress in Mdx Mice. International Journal of Molecular Sciences, 21(15), 5415.

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Immunocytochemical Profiling of Glial Cells

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Immunocytochemistry was performed as described earlier (20 (link)). Briefly, 8-well chamber slides containing BV-2 microglial cells, mouse primary microglia or astrocytes cultured to 70–80% confluence were fixed with chilled methanol (Fisher Scientific, Waltham, MA) overnight, followed by two brief rinses with filtered PBS. Samples were blocked with 2% BSA (Fisher Scientific) in PBS containing Tween 20 (Sigma) and Triton X-100 (Sigma) for 30 min and incubated at room temperature under shaking conditions for 2 hr in PBS containing the following anti-mouse primary antibodies: SOCS3 (1:200; Santa Cruz Biotech, Santa Cruz, CA), phospho-CREB (1:600; Cell Signaling, Beverly, MA), total CREB (1:600; Cell Signaling), GFAP, (1:100; Santa Cruz), and IBA1 (1:500; Cedarlane Laboratories, Burlington, NC). After four 15 min washes in filtered PBS, slides were incubated with Cy2 and Cy5-labeled secondary antibodies (all 1:200; Jackson ImmunoResearch, West Grove, PA) for 1 hr under shaking conditions. Following four 15 minute washes with filtered PBS, cells were incubated for 4–5 min with 4′,6-diamidino-2-phenylindole (DAPI, 1:10,000; Sigma). The samples were run in an ethanol and Xylene (Fisher) gradient, mounted and visualized under BX-41 Olympus fluorescence microscope (21 (link)–23 (link)).

Chakrabarti S., Jana M., Roy A, & Pahan K. (2018). Upregulation of suppressor of cytokine signaling 3 in microglia by cinnamic acid. Current Alzheimer research, 15(10), 894-904.

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Quantitative Analysis of Cellular Apoptosis

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Cell apoptosis was analyzed using Hoechst 33258 staining. SNU-1 cells were seeded into 12-well plates and treated with different concentrations (0, 5, 10 and 20 µg/ml) of Nar for 24 h. The adherent cells were washed twice with phosphate-buffered saline. The cells were then stained with Hoechst 33258 for 5 min at room temperature and washed twice. The blue-stained nuclei was observed under the BX41 fluorescence microscope (magnification, ×100; Olympus Corporation). Images were captured and used to quantitatively analyze the apoptosis of cells using Image-Pro Plus analysis software 6.0 (Media Cybernetics, Inc.).

Xu C., Huang X., Huang Y., Liu X., Wu M., Wang J, & Duan X. (2021). Naringin induces apoptosis of gastric carcinoma cells via blocking the PI3K/AKT pathway and activating pro-death autophagy. Molecular Medicine Reports, 24(5), 772.

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SNU-1 Cell Apoptosis Analysis

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Cell apoptosis was analyzed using TUNEL staining. SNU-1 cells were plated onto 12-well plates for 24 h and treated with different concentrations (0, 5, 10 and 20 µg/ml) of Nar for 24 h. Then, apoptosis was evaluated using the TUNEL apoptosis assay kit and observed under the BX41 fluorescence microscope (magnification, ×100; Olympus Corporation). Images were captured and used to quantitatively analyze the apoptosis of cells using Image-Pro Plus analysis software 6.0 (Media Cybernetics, Inc.).

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Cell Labeling for Bone Consolidation

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To verify that the transplanted cells remained on the defect site and contributed to bone consolidation, the cells were labeled with Qdots^®, 655 nm emission, 450-615 nm excitation (Qtracker^® Cell Labeling Kit, GIBCO Life Technologies, São Paulo, SP, Brazil). The labeling is provided by the presence of highly fluorescent nanocrystals in the cells’ cytoplasm. To confirm the label, 10 μL of the cells subjected to labeling were placed under a histological slide and coverslip and examined with a BX41 fluorescence microscope (Olympus, Shinjuku, Tokyo, Japan).

SOARES I.M., FERNANDES G.V., CAVALCANTE L.C., LEITE Y.K., BEZERRA D.D., de CARVALHO M.A, & CARVALHO C.M. (2019). The influence of Aloe vera with mesenchymal stem cells from dental pulp on bone regeneration: characterization and treatment of non-critical defects of the tibia in rats. Journal of Applied Oral Science, 27, e20180103.

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Detecting Extracellular Matrix Components

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Monolayers were grown on glass coverslips in 24-well polystyrene plates with 1 ml of H-broth for six days at 23 °C. They were washed twice with PBS and fixed with 3.8% formaldehyde in PBS for 30 min and again washed twice with PBS. The material was stained with 1 μl of 4′,6-diamidino-2-phenylindole (DAPI, 100 μg/ml in water) and 8 μl of Texas Red-conjugated concanavalin A (1 mg/ml) in 200 μl of PBS. While DAPI stains nucleic acids, concanavalin A (Con A) binds to glucosyl and mannosyl residues of the polysaccharides in the ECM. Stained films were imaged using an Olympus BX41 Fluorescence Microscope in the College of Arts and Sciences Imaging Center at Texas Tech University.

Silva S., Matz L., Elmassry M.M, & San Francisco M.J. (2019). Characteristics of monolayer formation in vitro by the chytrid Batrachochytrium dendrobatidis. Biofilm, 1, 100009.

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Proteinase K Digestion for α-Synuclein Immunostaining

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Proteinase K (PK) digestion of nigral sections was carried out during immunofluorescence staining of α-syn in dopaminergic neurons. The sections were kept in solution containing 10 μg/mL of PK made in PBS for 10 min at 37°C under constant rotation. Following the PK digestion, co-staining of α-syn and TH was performed using the protocol described above. Images were captured under Olympus BX41 fluorescence microscope and MFI was analyzed using ImageJ.

Dutta D., Paidi R.K., Raha S., Roy A., Chandra S, & Pahan K. (2022). Treadmill exercise reduces α-synuclein spreading via PPARα. Cell reports, 40(2), 111058.

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Fluorescence Microscopy for Amyloid Detection

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Ultraviolet evaluation was performed on all H&E and CR stained slides, and tissues were assessed for the presence or absence of autofluorescence. Ultraviolet evaluation has been shown to be a simple method to enhance CR evaluation of amyloid deposits (Sen and Basdemir 2003 (link)). The evaluation was performed utilizing an Olympus BX41 fluorescence microscope. Two filters were used including the “green” HG.TR (Texas Red) filter set (Chroma #41004) which has an absorption maximum of 560 nm (green) and an emission maximum of 645 nm (red), and the “blue” U-MNIB filter set (Olympus FITC (fluorescein isothiocyanate)) which has an absorption maximum of 470–490 nm (blue) and an emission maximum of 515 nm (green).

Hoane J.S., Johnson C.L., Morrison J.P, & Elmore S.A. (2016). Comparison of Renal Amyloid and Hyaline Glomerulopathy in B6C3F1 Mice: An NTP Retrospective Study. Toxicologic pathology, 44(5), 687-704.

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Immunofluorescent Staining of DJ-1 and TH

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It was performed as described earlier [50 (link),53 (link)]. Briefly, cover slips containing 100–200 cells/mm² were fixed with 4% paraformaldehyde followed by treatment with cold ethanol and two rinses in phosphate-buffered saline (PBS). Samples were then blocked with 3% bovine serum albumin (BSA) in PBS-Tween-20 (PBST) for 30 min followed by incubation in PBST containing 1% BSA and mouse anti-DJ-1 (1:200) or rabbit anti-TH (1:500). After three washes in PBST (15 min each), slides were further incubated with Cy2 (Jackson ImmunoResearch Laboratories, Inc.). For negative controls, a set of culture slides was incubated under similar conditions without the primary antibodies. The samples were mounted and observed under an Olympus BX41 fluorescence microscope.

Jana M., Dasarathy S., Ghosh S, & Pahan K. (2023). Upregulation of DJ-1 in Dopaminergic Neurons by a Physically-Modified Saline: Implications for Parkinson’s Disease. International Journal of Molecular Sciences, 24(5), 4652.

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