Bx ucb microscope

Manufactured by Olympus

Sourced in Japan

The BX-UCB microscope is a high-performance research-grade microscope designed for a variety of laboratory applications. It features a sturdy and ergonomic design, providing a stable platform for accurate and precise observations. The BX-UCB is equipped with advanced optics and illumination systems to deliver clear and detailed images.

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BX microscope (31 citations) BX-UCB (19 citations) BX-UCB fluorescent microscope (2 citations) BX-UCB/BX61 microscope (2 citations) BX-UCB light-field microscope (2 citations)

9 protocols using bx ucb microscope

ER Staining of Blastocysts

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Image analysis for ER staining was achieved by seeding cells on a coverslip-loaded six-well plate at 50 blastocysts/ml. At 16 h after plating, blastocyst were treated with CSNPs. Twelve hours later, the ER-Tracker Blue-White DPX (Molecular Probes, Eugene, OR) probe was added to blastocyst and was incubated for 30 min under the same growth conditions. The loading solution was removed, and blastocysts were then washed with PBS. Microscopic images were collected using an Olympus BX-UCB microscope and processed using DP controller software. The quantitative analysis of fluorescent intensity was performed using MeTaMorph image analysis software (Molecular Devices, California, USA).

Choi Y.J., Gurunathan S., Kim D., Jang H.S., Park W.J., Cho S.G., Park C., Song H., Seo H.G, & Kim J.H. (2016). Rapamycin ameliorates chitosan nanoparticle-induced developmental defects of preimplantation embryos in mice. Oncotarget, 7(46), 74658-74677.

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Immunofluorescence Analysis of Oocyte Cytoskeleton

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Ten oocytes were fixed in 4% paraformaldehyde for 30 min and permeabilised for 30 min with PBS containing 0.1% Triton X-100. Permeabilised oocytes were blocked for 1 h at room temperature in 1% bovine serum albumin (BSA) and 0.1% Triton X-100 in PBS before overnight incubation at 4 °C with the primary antibodies for anti -α-tubulin (Cell Signaling Technology, Beverly, MA, USA), anti-HDAC6 and anti-acetyl-α-tubulin. The oocytes were washed several times in 0.05% Tween 20 in PBS (PBST), transferred to a secondary antibody mixture of Alexa Fluor 568 goat anti-mouse and Alexa Fluor 488 goat anti rabbit (Molecular Probes, USA), and incubated at room temperature for 30 min. Aggregates of ubiquitinated proteins (aggresomes) were detected in oocytes after treatment with Tub A using a ProteoStat Aggresome Detection Kit (Enzo Life Sciences, Inc., USA), and confocal images using the TO-PRO-3 fluorescent dye were acquired using an Olympus FV1000 Confocal microscope (Tokyo, Japan), and were processed using the FV10-ASW 2.0 Viewer software (Olympus, Tokyo, Japan). Fluorescent images were acquired using an Olympus BX-UCB microscope and were processed using a DP controller software (Olympus, Tokyo, Japan). The quantitative analysis of the fluorescent intensity was performed using the MeTaMorph image analysis software (Molecular Devices, California, USA).

Zhou D., Choi Y.J, & Kim J.H. (2017). Histone deacetylase 6 (HDAC6) is an essential factor for oocyte maturation and asymmetric division in mice. Scientific Reports, 7, 8131.

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Collagen Matrix Cell Viability Assay

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Cell viability was quantified and assessed using a live/dead reduced biohazard viability/cytotoxity kit (Molecular Probes, L-3224), following the manufacturer's protocol. Live and dead cells were simultaneously identified with calcein AM (green) and ethidium homodimer (red) stains, respectively. The collagen constructs were unrolled using forceps and surgical blades as described above. The segments were placed into culture wells (12-well) and covered with 0.5 ml of the live/dead solution (20 μl of calcein AM/5 ml PBS added to 17 μl of ethidium homodimer/5 ml PBS, which equates to 2 μM calcein AM and 8 μM ethidium homodimer). The collagen matrices were incubated in live/dead solution at 37°C for 45 min before being mounted onto slides and viewed under an Olympus BX-UCB microscope. The green fluorescent nucleic acid and dead red stain were used to differentiate live and dead cells, respectively. Images were captured by the microscope and live/dead nuclei counted to ascertain percentage viability. Nine randomly selected areas within the outer, middle and core were analysed, resulting in 27 randomly selected areas being captured and counted within each construct. Data presented are live cells as a percentage of total cell number (live + dead).

Ardakani A.G., Cheema U., Brown R.A, & Shipley R.J. (2014). Quantifying the correlation between spatially defined oxygen gradients and cell fate in an engineered three-dimensional culture model. Journal of the Royal Society Interface, 11(98), 20140501.

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Immunofluorescence Analysis of Serotonin in Colon

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Immunofluorescence labelling was performed on 10 μm transverse sections of distal colon. Tissues were fixed in 4% paraformaldehyde in PBS and then embedded in paraffin. Sections were deparaffinised in Histoclear and rehydrated in graded ethanol solutions. The slides were then incubated for 30 min in 2 N HCl for antigen retrieval. The sections were then incubated with 10% goat serum (S-1000; Vector lab, Peterborough, UK) for 2 hours at room temperature. Rabbit anti-serotonin primary antibody (1:8000; 20080; ImmunoStar, UK) was applied overnight at 4 °C, and then slides were washed three times with PBS (10 min each wash). Sections were incubated for 2 hours with Alexa 488 goat anti-rabbit secondary antibody (1:200, A-11034; Invitrogen, UK). After washing with PBS, sections were mounted with Fluoroshield DAPI (F6057; Sigma, UK). Sections were viewed and images were acquired using an Olympus BX-UCB microscope.

Patel B.A., Fidalgo S., Wang C., Parmar L., Mandona K., Panossian A., Flint M.S., Ranson R.N., Saffrey M.J, & Yeoman M.S. (2017). The TNF-α antagonist etanercept reverses age-related decreases in colonic SERT expression and faecal output in mice. Scientific Reports, 7, 42754.

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Exploring GO-AgNPs Effects on CFFCs

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CFFCs were seeded in a 24-well plate for 24 h and then treated with 0, 4, and 8 μg/mL of GO-AgNPs for 24 h. Cell morphology was observed using an Olympus BX-UCB microscope (Tokyo, Japan).

Yuan Y.G., Cai H.Q., Wang J.L., Mesalam A., Md Talimur Reza A.M., Li L., Chen L, & Qian C. (2021). Graphene Oxide–Silver Nanoparticle Nanocomposites Induce Oxidative Stress and Aberrant Methylation in Caprine Fetal Fibroblast Cells. Cells, 10(3), 682.

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Mitochondrial Calcium Dynamics Visualization

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A Rhod2-AM (Molecular Probes, Eugene, OR) probe was used to determine the mitochondrial Ca²⁺ level [68 (link)]. Rhod2-AM possesses a net positive charge, which facilitates its sequestration into the mitochondria because of its membrane potential-driven uptake. The use of Rhod2-AM enhances the selectivity for mitochondrial loading because this dye exhibits Ca²⁺-dependent fluorescence only after it is oxidized, which occurs preferentially within the mitochondria. Cells were treated with CSNPs and incubated for 12h, and the cells were then harvested, washed, and suspended in PBS containing Rhod2-AM (1μM). For image analysis, the cells were loaded with Rhod2-AM and incubated for 30 min at 37°C. Cells were washed, and the stained cells were mounted onto a microscope slide with mounting medium (Vector Laboratories, USA). For mitochondrial staining, embryos in culture were incubated in 160nM MitoTracker (Molecular Probes, OR, USA) in DMEM before being fixed and immunostained. For green fluorescent protein (GFP) visualization, cells were only fixed and washed. Microscopic images were collected using an Olympus BX-UCB microscope and were processed using DP controller software. Quantitative analysis of fluorescence intensity was performed using MeTaMorph image analysis software (Molecular Devices, California, USA).

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Quantifying Neural Activity in Raphe and Amygdala

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DAB- and immunofluorescently-labeled tissue was examined using an Olympus BX-UCB microscope (http://www.olympusamerica.com/) equipped with a motorized stage (96S100-LE; Ludl Electronic Products, http://www.ludl.com/), fluorophore-specific fluorescent filter sets (excitation and emission spectra: Cy3/TRITC – 531/40), and a cooled mono CCD camera (Orca R2; Hamamtsu, http://hamamatsucameras.com/). Tissue sections through the dorsal raphe and amygdala were examined at 240 µm intervals. In the dorsal raphe, sections were divided into multiple subregions: caudal dorsomedial dorsal raphe (cDRD), ventrolateral wings of the dorsal raphe (DRVL), dorsomedial dorsal raphe (DRD), ventral dorsal raphe (DRV), rostral dorsomedial dorsal raphe (rDRD), and rostral ventral dorsal raphe (rDRV) [15 (link)]. The number of cells dual-labeled with c-Fos and Tph2 were counted in each subregion through the extent of the dorsal raphe (Fig. 2A). In the amygdala, the following subnuclei were examined: lateral amygdala (LA), basolateral amygdala (BLA), central amygdala (CeA), medial amygdala (MeA), and basomedial amygdala (BMA). All c-Fos labeled cells were counted throughout the rostrocaudal extent of each nucleus in 240 µm increments (Fig. 3A). Figures were prepared using Adobe Photoshop CS5 (http://www.adobe.com/); brightness and contrast were optimized for presentation purposes.

Cohen J.L., Ata A.E., Jackson N.L., Rahn E.J., Ramaker R.C., Cooper S., Kerman I.A, & Clinton S.M. (2016). Differential stress induced c-Fos expression and identification of region-specific miRNA-mRNA networks in the dorsal raphe and amygdala of high-responder/low-responder rats. Behavioural brain research, 319, 110-123.

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Apoptosis Detection in Ovary Tissues

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Apoptosis was assessed using the In Situ Cell Death Detection Kit (Roche, West Sussex, UK) and 4',6-diamidino-2-phenylindole (DAPI) dual staining (Vector Laboratories, CA, USA). Briefly, the cells and ovary tissues in each group were prepared, washed twice with D-PBS and then incubated with TUNEL reaction mixture for 60 min at 37°C. The cells undergoing apoptosis were detected using an Olympus BX-UCB microscope and were processed using DP controller software. At least 20 blastocysts were analyzed for each determination.

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Circadian Regulation of PER2 in Mouse Brain

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A separate group of 5-month-old SAMP8 and SAMR1 male mice were sacrificed every 4 hours at ZT 3, 7, 11, 15, 19, and 23. Brains were harvested, fixed in 4% paraformaldehyde at 4° C overnight and cryoprotected in 20% sucrose. Slices (40 μm thick) were sectioned on a cryostat. Free-floating slices were blocked in 5% bovine serum albumin, 0.3% Triton-X in phosphate buffered saline at room temperature for 1-h and then incubated with rabbit anti-PERIOD2 (PER2; 1:2,000; EDM Millipore) overnight at 4° C. Slices were incubated in goat anti-rabbit Alexafluor 488 (1:500; Invitrogen) secondary for 2-h at room temperature. Slices were mounted onto slides with VECTASHIELD HardSet mounting media with DAPI (Vector Laboratories). Slides were imaged on an Olympus BX-UCB microscope at consistent exposure time, gain, and contrast settings. Fluorescence intensity (arbitrary units) was measured using Image J software (NIH) as previously described (Paul et al., 2017 (link)) by drawing a standard region of interest for the whole SCN. Background intensity was measured from the anterior hypothalamus outside the SCN and subtracted from all values. Representative hippocampal images were provided to visualize the location of the PER2 signal in the hippocampus but were not quantified.

Davis J.A., Paul J.R., Mokashi M.V., Yates S.A., Mount D.J., Munir H.A., Goode L.K., Young M.E., Allison D.B, & Gamble K.L. (2022). Circadian disruption of hippocampus in an early senescence male mouse model. Pharmacology, biochemistry, and behavior, 217, 173388.

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