Bx61 upright wide field microscope

Manufactured by Olympus

The BX61 is an upright wide field microscope manufactured by Olympus. It is designed for a variety of microscopy applications. The microscope features a sturdy construction and a wide field of view to enable efficient observation and documentation of samples.

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Volocity software, by PerkinElmer (1 mentions)

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BX61 microscope (746 citations) Upright microscope (110 citations) BX61 upright microscope (36 citations) BX61 wide-field microscope (4 citations)

4 protocols using bx61 upright wide field microscope

Oil Red O Staining of Armadillo Liver

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Oil red O (ORO) staining was performed on frozen 10 μm Armadillo liver sections, according to published methods.⁸⁶ (link) As a positive control, sections of steatotic human liver were used. Slides were left to equilibrate for 10 min at room temperature, covered with filtered ORO working solution for 5 min, before washing for 30 min with running water. Slides were mounted with Fluorsave and glass coverslips and sealed with nail polish prior to imaging on an Olympus BX61 upright widefield microscope with a 10x objective lens.

Hess S., Kendall T.J., Pena M., Yamane K., Soong D., Adams L., Truman R, & Rambukkana A. (2022). In vivo partial reprogramming by bacteria promotes adult liver organ growth without fibrosis and tumorigenesis. Cell Reports Medicine, 3(11), 100820.

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Quantifying DNA Damage Responses in pMMECs

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pMMECs were infected twice with either Cre-sgControl, Cre-sgMre11, GFP, or NeuT virus. Upon confirmation of at least 80% viral efficiency, cells were seeded onto 3D-matrix coated coverslips, were treated with EdU for 10 min, and subsequently fixed by cold Methanol:Acetone (1:1) incubation at −20Cfor 10 mins. Cells then underwent EdU detection using the EdU detection kit (Baseclick) in accordance with kit instructions. Cells were then blocked in PBS + 5% FBS for 1 hour, followed by incubation in the appropriate primary antibody for 1 hour (p-γH2AX; 53bp1; p-RPA2; or S9.6), secondary antibody for 30 min, then DAPI for 1 min. Coverslips were then mounted onto slides with Prolong Gold mounting medium, cured for 2 hours and stored at 4C in the dark until imaging. Coverslips were examined on an Olympus BX61 upright wide field microscope. Resulting foci were analyzed using Fiji software (Schindelin etal., 2012 (link)).

Fagan-Solis K.D., Simpson D.A., Kumar R.J., Martelotto L.G., Mose L.E., Rashid N.U., Ho A.Y., Powell S.N., Wen Y.H., Parker J.S., Reis-Filho J.S., Petrini J.H, & Gupta G.P. (2020). A P53-Independent DNA Damage Response Suppresses Oncogenic Proliferation and Genome Instability. Cell reports, 30(5), 1385-1399.e7.

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Comet Assay for DNA Damage

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pMMECs were infected twice with either Cre-sgControl, Cre-sgMre11, GFP, or NeuT virus and confirmation of at least 80% viral efficiency was determined by Flow Cytometry (Attune NxT) for GFP expression. The presence of SSBs and DSBs were analyzed via Alkaline (SSBs/DSBs) and Neutral (DSBs) comet assay using the Trevigen comet assay kit according to manufacturer’s protocol. Comet images were captured by fluorescence microscopy using Olympus BX61 upright wide field microscope. The tail DNA percent was quantified using the ImageJ software with OpenComet plug-in (Gyori et al., 2014 (link)).

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Fluorescence Microscopy for γH2AX Quantification

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Fluorescence microscopy was performed using an Olympus BX61 upright wide field microscope with Hg lamp. Images were captured using a Hamamatsu ORCA-R2 (C10600–10B) camera and Improvision’s Volocity Software. Separate grayscale images were recorded for Hoescht (Nuclei, blue) and AlexaFluor 594 (γH2AX, red) - excitation(ex) and emission(em) filters were as follows: 1) DAPI (blue) ex: 377/50 em: 447/60 and 2)TxRed (red) ex: 562/40; em: 624/40. Images were acquired with 20X/0.50 UPlanFLN and 60X/1.42 Oil PLanApo N objective lenses. Imaging parameters were set to avoid saturation of pixels and exposure was optimized to prevent photobleaching. All images in a single biological replicate, containing essential controls, were collected with the same acquisition settings. Six randomized images were taken per each technical replicate. In total, we collected images from 3–4 biological replicates with 1–2 technical replicates per sample group (n = 3–4). %γH2AX⁺ cells were quantified based on equation (2).

% γ H 2 A X^{+} cells = [\frac{number of cells with > 3 γ H 2 A X foci}{number of nuclei}] \times 100

Sadecki P.W., Balboa S.J., Lopez L.R., Kedziora K.M., Arthur J.C, & Hicks L.M. (2021). Evolution of polymyxin resistance regulates colibactin production in Escherichia coli. ACS chemical biology, 16(7), 1243-1254.

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