Bx51 widefield microscope

Manufactured by Olympus

The BX51 widefield microscope is a high-quality optical instrument designed for various scientific and research applications. It features a sturdy, ergonomic design and offers a wide range of magnification capabilities to facilitate detailed observations and analyses. The BX51 is equipped with advanced optics and illumination systems to provide clear, high-contrast images. This microscope is suitable for a variety of sample types and can be utilized in diverse laboratory settings.

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Lab products found in correlation

Sybr gold, by Thermo Fisher Scientific (2 mentions) Low melting point agarose, by Merck Group (2 mentions) Tris borate edta buffer, by Merck Group (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Ez c1 freeviewer, by Nikon (1 mentions) Te2000 psf inverted microscope, by Nikon (1 mentions)

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BX51 microscope (3784 citations)

7 protocols using bx51 widefield microscope

Alkaline Comet Assay for DNA Damage

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Cells were trypsinized and frozen in 10% DMSO in fetal bovine serum and stored at −80°C prior to downstream processing. Cells were washed in cold PBS and resuspended in 0.7% Low Melting Point agarose (Sigma, A9414) at 37°C to a concentration of 2 × 10⁵ cells/ml. Cell/agarose mix (70 μl) was placed on slides previously coated in 1% agarose. Slides were incubated in lysis buffer (2.5 M NaCl, 100 nM EDTA, 10 nM Tris, pH 10, 250 nM NaOH, 10% DMSO, 1% Triton X-100) for 1 h at 4°C. Slides were then washed twice in cold PBS.
Samples were subjected to electrophoresis for 30 min at 25 V at 4°C in Tris-borate EDTA buffer (Sigma, T3913). Slides were then washed twice in cold PBS and stained with Sybr Gold (Life Technologies, S11494) in Tris-borate EDTA buffer for 45 min. Slides were washed twice in MilliQ water and allowed to dry. Samples were imaged using an Olympus BX51 widefield microscope with Olympus UPlanFL 20x/0.50 air objective. Comets were scored using Comet assay IV (Perceptive Instruments). For each sample, 100 randomly captured comets (50 cells on each of 2 comet slides) were quantified.

Hewitt G., Carroll B., Sarallah R., Correia-Melo C., Ogrodnik M., Nelson G., Otten E.G., Manni D., Antrobus R., Morgan B.A., von Zglinicki T., Jurk D., Seluanov A., Gorbunova V., Johansen T., Passos J.F, & Korolchuk V.I. (2016). SQSTM1/p62 mediates crosstalk between autophagy and the UPS in DNA repair. Autophagy, 12(10), 1917-1930.

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Visualizing DDR1 Receptor Localization

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Cos-7 cells transiently expressing the ectodomain deletion constructs, DDR1b-ΔECD, DDR1b-ΔECD-TM1, DDR1b-ΔECD-Cys or DDR1b-ΔECD-Cys-TM1, which contain N-terminal Flag tags, were washed with PBS, and incubated with mouse anti-Flag IgG1 for 1 hr on ice to label cell surface DDR1 constructs. This was followed by washing, fixing with 4% paraformaldehyde (Sigma) in PBS for 15 min at room temperature, blocking in 8% BSA in PBS for 1 hr, and incubation with anti-mouse IgG1 Alexa-Fluor-488 secondary Ab. Cos-7 cells transiently expressing DDR1b wildtype were stimulated with 10 µg/ml collagen I or a mixture of 10 µg/ml collagen and 10 µg/ml mouse anti-DDR1 7A9 IgG1 mAb, for 10 min at 37°C. The cells were washed with PBS, and incubated with mAb 7A9 for 1 hr on ice to label cell surface DDR1. This was followed by fixation and incubation with secondary Ab as above. The coverslips were mounted on slides with Prolong Gold Antifade Reagent (Invitrogen). The cells were examined using an Olympus BX-51 widefield microscope.

Juskaite V., Corcoran D.S, & Leitinger B. (2017). Collagen induces activation of DDR1 through lateral dimer association and phosphorylation between dimers. eLife, 6, e25716.

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Comet Assay for Genotoxicity Evaluation

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Cells were trypsinized and frozen in 10% DMSO in fetal bovine serum and stored at −80°C prior to downstream processing. Cells were washed in cold PBS and resuspended in 0.7% Low Melting Point agarose (Sigma, A9414) at 37°C to a concentration of 2 × 10⁵ cells/ml. Cell/agarose mix (70 μl) was placed on slides previously coated in 1% agarose. Slides were incubated in lysis buffer (2.5 M NaCl, 100 nM EDTA, 10 nM Tris, pH 10, 250 nM NaOH, 10% DMSO, 1% Triton X‐100) for 1 h at 4°C. Slides were then washed twice in cold PBS. Samples were subjected to electrophoresis for 40 min at 25 V at 4°C in alkaline buffer (300 mM NaOH, 1 mM EDTA). Slides were then washed twice in cold PBS and stained with Sybr Gold (Life Technologies, S11494) in Tris‐borate EDTA buffer for 40 min. Slides were washed twice in MilliQ water and allowed to dry. Samples were imaged using an Olympus BX51 widefield microscope with Olympus UPlanFL 20×/0.50 air objective. Comets were scored using Comet assay IV (Perceptive Instruments). For each sample, 100 randomly captured comets (50 cells on each of 2 comet slides) were quantified.

Lagnado A., Leslie J., Ruchaud‐Sparagano M., Victorelli S., Hirsova P., Ogrodnik M., Collins A.L., Vizioli M.G., Habiballa L., Saretzki G., Evans S.A., Salmonowicz H., Hruby A., Geh D., Pavelko K.D., Dolan D., Reeves H.L., Grellscheid S., Wilson C.H., Pandanaboyana S., Doolittle M., von Zglinicki T., Oakley F., Gallage S., Wilson C.L., Birch J., Carroll B., Chapman J., Heikenwalder M., Neretti N., Khosla S., Masuda C.A., Tchkonia T., Kirkland J.L., Jurk D., Mann D.A, & Passos J.F. (2021). Neutrophils induce paracrine telomere dysfunction and senescence in ROS‐dependent manner. The EMBO Journal, 40(9), e106048.

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Confocal Microscopy of Spheroids

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Spheroids were prepared for confocal microscopy as described 29 (link). Details of the antibodies used are given in Supporting Information Table S2. Images were collected on a Nikon C1 confocal using a TE2000 PSF inverted microscope, using ×60 /NA 1.40 Plan Apo or ×20/NA 0.50 Plan Fluor objectives and ×3 confocal zoom. Different sample images detecting the same antibodies were acquired under constant acquisition settings. Images were processed using Nikon EZ-C1 FreeViewer v3.3 software. Bright-field images were collected on an Olympus BX51 widefield microscope, using a ×10/NA 0.3 UPlan F1 objective. Images were captured with a CoolSNAP camera system and processed using MetaMorph imaging v5.0 software.

Ball S.G., Worthington J.J., Canfield A.E., Merry C.L, & Kielty C.M. (2014). Mesenchymal Stromal Cells: Inhibiting PDGF Receptors or Depleting Fibronectin Induces Mesodermal Progenitors with Endothelial Potential. Stem Cells (Dayton, Ohio), 32(3), 694-705.

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In Vitro CXCR4-Targeted MB Binding

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For in vitro MB binding studies, MDA-MB-231 shSC or shCXCR4 cells (1x10⁶) were plated in OptiCell™ plates (VWR, Radnor, Pennsylvania, USA) in 10 mL of media. CXCR4 knockdown was induced by treatment with 0.5 μg/mL of DOX for 24h. Cells were incubated with approximately 1x10⁶ MB/mL (10 mL) of either NT-MB or T140-MB for 15 min at 37°C. During this period, the OptiCell™ plates were flipped to maximize contact between the cells and MB; after incubation, plates were flipped back to the original position and allowed to rest for 5 min before evaluation as to ensure floating of unbound MB to the top of the plate. Blocking studies were performed by incubation with 1 mg/mL of unconjugated T140 for 5 min before and during MB incubation. Fluorescence evaluation of MB binding was carried out as described, albeit with the use of Dil-modified MB. GFP-expressing MDA-MB-231-Fluc-GFP were plated in parallel with these experiments and used as control. Images were acquired using a 40x UPlanAPO objective lens on the Olympus BX-51 wide-field microscope referenced previously.

Braga M., Leow C.H., Gil J.H., Teh J.H., Carroll L., Long N.J., Tang M.X, & Aboagye E.O. (2021). Investigating CXCR4 expression of tumor cells and the vascular compartment: A multimodal approach. PLoS ONE, 16(11), e0260186.

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Mitochondrial Redox and pH Dynamics Imaging

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To record mitochondrial redox and pH dynamics in motor axons and NMJs, we used a BX51 wide-field microscope (Olympus) equipped with ×20/0.5 NA and ×100/1.0 NA dipping-cone water-immersion objectives, a cooled charged-coupled device camera and a PolyV polychromator system (Till Photonics, controlled by TillVision software; dichroic filter: D/F 500 DCXR; emission filter: ET 525/36). Images were acquired at rates of 1 Hz with exposure times of 150 (450) ms for 408 nm excitation and 30 (150) ms for 488 nm excitation. Some of the axon movies have been previously manually analyzed and published in the context of single contractions8 (link).

Breckwoldt M.O., Armoundas A.A., Aon M.A., Bendszus M., O’Rourke B., Schwarzländer M., Dick T.P, & Kurz F.T. (2016). Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters. Scientific Reports, 6, 23251.

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Rice Blast Infection and Penetration Assays

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For blast infection assay, conidial suspension (10⁶/mL) with 0.01% gelatine was sprayed on 21‐day‐old rice seedlings (Oryza sativa cultivar CO39) and incubated in a growth chamber (16 h light/d, 22 °C and 90% humidity). Blast disease in infection assays was assessed and recorded by scanning the leaves at 7 dpi. The blast infection assays were repeated at least three times.
For the host penetration and in planta invasive hyphal development assay, healthy rice seedlings (CO39) at the age of 4 weeks were selected for sheath preparation. Conidial suspension (5 × 10⁴/mL) were inoculated onto rice sheath and incubated on sterile wet tissue paper in 90 mm Petri dishes. The Petri dishes with inoculated rice sheaths were transferred into the growth chamber with a photoperiod of 16 h:8 h light:dark cycle at 25 °C. The inoculated sheath was trimmed manually and observed by using an Olympus BX51 wide field microscope or with a laser scanning confocal microscope at selected time points.
To prepare heat‐killed rice sheaths, the fresh rice sheaths were immersed into sterile water at 70 °C for 25 min (Shipman et al., 2017). The heat‐killed rice sheath has the physical structures of cells, while the abilities of host response to fungal infection are lost.

Kou Y., He Y., Qiu J., Shu Y., Yang F., Deng Y, & Naqvi N.I. (2019). Mitochondrial dynamics and mitophagy are necessary for proper invasive growth in rice blast. Molecular Plant Pathology, 20(8), 1147-1162.

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