Bx61wi

Manufactured by Olympus

Sourced in Japan, United States

The BX61WI is a motorized upright microscope system designed for a variety of imaging applications. It features a high numerical aperture objective lens, a motorized XY stage, and a sensitive camera interface for capturing high-quality images.

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Lab products found in correlation

Fv1000mpe2, by Olympus (6 mentions) Xlplann25xw, by Spectra-Physics (6 mentions) Maitai hp deepsee, by Spectra-Physics (5 mentions) Multiclamp 700b amplifier, by Molecular Devices (4 mentions) Lsm 880, by Olympus (3 mentions) Zen black edition software, by Zeiss (3 mentions) Fluoview fv1000mpe, by Olympus (3 mentions) Pclamp 10, by Molecular Devices (3 mentions) Fv1000mpe, by Olympus (2 mentions) C9100 13, by Hamamatsu Photonics (2 mentions) Pkh26 linker kits, by Merck Group (2 mentions) Prairie view 5, by Bruker (2 mentions) Ti sapphire pulsed laser system, by Spectra-Physics (2 mentions) Digidata 1550, by Molecular Devices (2 mentions) Digidata 1322a, by Molecular Devices (2 mentions) Fv1000, by Olympus (2 mentions) Axio observer z1 7 wide, by Zeiss (2 mentions) Maitai, by Spectra-Physics (2 mentions) Donkey anti mouse secondary antibody, by Jackson ImmunoResearch (2 mentions) Rabbit anti tph, by Merck Group (2 mentions)

61 protocols using bx61wi

Zinc Uptake in Human Epithelial Cells

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Human epithelial cells Hs27 (ATCC-CRL-1634) were seeded in 6-well plates at a density of 1 × 10⁵ cells per well in 2 mL of Dulbecco's Modified Eagle Media (DMEM, Invitrogen Life Technologies) supplemented with 10% fetal bovine serum (Gibco Life Technologies) 100 IU mL⁻¹ of penicillin, and 100 g mL⁻¹ of streptomycin. Cells were maintained at 37 °C in a controlled humid atmosphere of 5% CO₂ and 95% air. Twenty-four hours later the medium was renewed and cells were loaded with Zn²⁺ at concentrations of 100 μM (37 °C for 60 min). After removal of free Zn²⁺ by washing with media, cells were exposed to the compounds (20 μM) and incubated for 30 minutes, and finally washed with PBS. Untreated cells were used as controls. Fluorescence images were collected using a confocal laser microscopy (Olympus BX61WI).

Berrones-Reyes J.C., Muñoz-Flores B.M., Cantón-Diáz A.M., Treto-Suárez M.A., Páez-Hernández D., Schott E., Zarate X, & Jiménez-Pérez V.M. (2019). Quantum chemical elucidation of the turn-on luminescence mechanism in two new Schiff bases as selective chemosensors of Zn2+: synthesis, theory and bioimaging applications. RSC Advances, 9(53), 30778-30789.

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Visualizing Neuronal Interactions with Multiphoton Microscopy

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Images of cells were visualized with an upright microscope (BX61WI, Olympus) equipped with a water immersion objective lens (XLPlan N 25X/NA 1.05w, Olympus) and the laser scanning microscope system (FV1000-MPE, Olympus), which was coupled with a mode-locked Ti:sapphire laser (MaiTai HP DeepSee, Spectra-physics, Mountain View, CA). The mode-locked lasers were set at the wavelength of 800 and 930 nm for excitation of Alexa 594 which was injected into target neurons and Venus/YFP which were expressed in a given type of interneurons, respectively. Emitted fluorescence was divided into long wavelength (>570 nm) and short wavelength light with a dichroic mirror (570 nm, Olympus), and short-wavelength light was further filtered through a bandpass filter (510–550 nm, Olympus). Both wavelengths of emitted fluorescence were detected simultaneously using two photomultiplier tube detectors (PMTs). The microscope objective was shielded from possible stray light by covering the space over the animal’s head with lightproof cloth and clay.

Safari M.S., Mirnajafi-Zadeh J., Hioki H, & Tsumoto T. (2017). Parvalbumin-expressing interneurons can act solo while somatostatin-expressing interneurons act in chorus in most cases on cortical pyramidal cells. Scientific Reports, 7, 12764.

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Probing Cell-Particle Interactions

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HUVECs were stained with Fura-2 AM and washed with a HEPES-buffered Tyrode’s solution. Mesostructured and solid Si particles have similar lateral sizes of ~5 μm. Particles sitting on cell bodies were pushed by glass micropipettes controlled by a micromanipulator. After making contact with the particles, pipettes were lowered by another 1 μm and held still for 20 s before retraction. During the whole process, fluorescence images were collected using an upright microscope (BX61WI, Olympus, Japan) equipped with an EM-CCD camera (C9100-13, Hamamatsu Photonics, Japan).

Jiang Y., Carvalho-de-Souza J.L., Wong R.C., Luo Z., Isheim D., Zuo X., Nicholls A.W., Jung I.W., Yue J., Liu D.J., Wang Y., De Andrade V., Xiao X., Navrazhnykh L., Weiss D.E., Wu X., Seidman D.N., Bezanilla F, & Tian B. (2016). Heterogeneous silicon mesostructures for lipid-supported bioelectric interfaces. Nature materials, 15(9), 1023-1030.

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Slice Culture Calcium Imaging

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Slice cultures were transferred to the recording chamber mounted on an upright microscope (Olympus BX61WI) equipped with a FluoView300 confocal laser-scanning system (Olympus, Tokyo, Japan) or on a two-photon microscope (Femto2D, Femtonics, Hungary) using a 40× water immersion objective (N.A. 0.80). Slices were superfused with oxygenated (95% O₂, 5% CO₂) artificial cerebrospinal fluid (ACSF) (5 ml/min, 30 °C), containing (in mM): 129 NaCl; 1.23 NaH₂PO₄; 10 glucose; 1.6 CaCl₂; 3 KCl; 21 NaHCO₃; 1.8 mM MgSO₄ (pH 7.4). In confocal microscopy GCaMP2 fluorescence was excited at 488 nm and emission was detected between 510–530 nm. In two-photon microscopy GCaMP2 was excited at 900 nm with a MaiTai femtosecond laser source (Spectra Physics, Santa Clara, CA, USA) and the emitted fluorescence was monitored at 475–575 nm.

Szabó Z., Héja L., Szalay G., Kékesi O., Füredi A., Szebényi K., Dobolyi Á., Orbán T.I., Kolacsek O., Tompa T., Miskolczy Z., Biczók L., Rózsa B., Sarkadi B, & Kardos J. (2017). Extensive astrocyte synchronization advances neuronal coupling in slow wave activity in vivo. Scientific Reports, 7, 6018.

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Confocal Microscopy Imaging Protocol

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Confocal images were taken on an Olympus BX61WI fixed stage microscope with Olympus Fluoview FV1000 confocal laser scanning. Images were processed in ImageJ.

Whittle J., Antunes L., Harris M., Upshaw Z., Sepich D.S., Johnson A.N., Mokalled M., Solnica‐Krezel L., Dobbs M.B, & Gurnett C.A. (2020). MYH3‐associated distal arthrogryposis zebrafish model is normalized with para‐aminoblebbistatin. EMBO Molecular Medicine, 12(11), e12356.

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In Vivo Two-Photon Imaging of Synaptic Markers

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This procedure has been described previously.⁸⁰ (link)^,⁸¹ (link) Briefly, Adeno-associated virus 1 (AAV1)-EGFP harboring the synapsin I promoter (titer: 1 × 10¹⁰ vector genomes/mL; 1 μL) and AAV2-VAMP2-mCherry harboring the CMV promoter were injected into adjacent positions in the frontal cortex (+1.0 mm from bregma (mediolateral 0.5 mm; depth, 1 mm; and +3.0 mm from bregma (mediolateral 0.5 mm; depth, 1 mm), respectively) under anesthesia with 1% isoflurane.
Two-photon imaging was performed using a laser-scanning microscope system FV1000MPE2 (Olympus, Tokyo, Japan) equipped with an upright microscope (BX61WI, Olympus, Japan), a water-immersion objective lens (XLPlanN25xW; numerical aperture, 1.05), and a pulsed laser (MaiTaiHP DeepSee, Spectra Physics, Santa Clara, CA, USA). EGFP and mCherry were excited at 920 nm and scanned at 495–540 nm and 575–630 nm, respectively. High-magnification imaging (101.28 μm × 101.28 μm; 1024 × 1024 pixels; 1 μm Z step) of cortical layer I was performed with a 5 × digital zoom through a thinned-skull window in the frontal cortex. Blinded observers performed image acquisition and analysis. Image processing was performed with Imaris Interactive Microscopy Image Analysis software (Bitplane, Zurich, Switzerland).

Shiwaku H., Katayama S., Kondo K., Nakano Y., Tanaka H., Yoshioka Y., Fujita K., Tamaki H., Takebayashi H., Terasaki O., Nagase Y., Nagase T., Kubota T., Ishikawa K., Okazawa H, & Takahashi H. (2022). Autoantibodies against NCAM1 from patients with schizophrenia cause schizophrenia-related behavior and changes in synapses in mice. Cell Reports Medicine, 3(4), 100597.

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Electrophysiological Analysis of NAc Neurons

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All experiments were performed blind to drug treatment or CSDS behavioral phenotype. Animals were perfused with ice-cold artificial cerebrospinal fluid (ACSF; see Supplementary Methods). Coronal slices (200 μm thick) containing NAc were cut with a microslicer, transferred for 1 hour to a holding chamber containing sucrose-ACSF (where 254 mM sucrose replaced NaCl) at 34°C and subsequently maintained at room temperature (20–22°C) until use. Individual slices were transferred to a recording chamber mounted on an upright microscope (Olympus BX61WI) and continuously perfused (2–3 ml per minute) with oxygenated ACSF also containing 3.5 mM MNI-glutamate (4-methoxy-7-nitroindolinyl-caged-L-glutamate; Tocris) and 10 μM D-serine (Sigma). Recordings were performed at room temperature. Whole-cell voltage-clamp recordings were made from D1-MSNs (EGFP-) and D2-MSNs (EGFP+) in the NAc shell region. Previous studies have established that EGFP⁻ cells in Drd2-EGFP mice reliably represent D1 MSNs in dorsal striatum (25 (link)), although we recognize that a clean separation of these two cell types is more ambiguous in the NAc (26 (link); 27 (link)). Cells were held at −70 mV. Recordings from EGFP⁺ and EGFP⁻ cells were interleaved.

Khibnik L.A., Beaumont M., Doyle M., Heshmati M., Slesinger P.A., Nestler E.J, & Russo S.J. (2015). Stress and Cocaine Trigger Divergent and Cell Type-Specific Regulation of Synaptic Transmission at Single Spines in Nucleus Accumbens. Biological psychiatry, 79(11), 898-905.

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Analyzing Autophagy in Fluorescently Labeled Cells

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Cells were plated onto poly-L-lysine coated round coverslips in a 6-well plate, grown overnight and subjected to experimental conditions as indicated. Stable cell lines expressing recombinant fluorescence proteins were imaged immediately after mounting on glass slides with ProLong™ Gold Antifade Mountant with DAPI (Thermo). For immunofluorescence, cells were fixed with 4% paraformaldehyde, briefly washed with PBS, and subsequently permeabilized with 0.5% Triton X-100 in PBS for 10 min followed by 1 h blocking in 2% BSA at RT. Coverslips were then incubated with anti-LC3A/B antibody (4108 S, CST; 1:250) for overnight at 4 °C, washed three times with PBS, and incubated again with DAPI and Alexafluor™ 594 conjugated anti-rabbit secondary antibody (A11012, Thermo; 1:250) for 1 h at RT. After 3 washes with PBS, coverslips were mounted on glass slides with ProLong™ Gold antifade reagent and examined under LSM510 META (Carl Zeiss) and BX61WI (Olympus) confocal microscopes. LC3-specific puncta were counted manually by ImageJ in minimum 25 cells per experimental group.

Maheshwari M., Yadav N., Hasanain M., Pandey P., Sahai R., Choyal K., Singh A., Nengroo M.A., Saini K.K., Kumar D., Mitra K., Datta D, & Sarkar J. (2022). Inhibition of p21 activates Akt kinase to trigger ROS-induced autophagy and impacts on tumor growth rate. Cell Death & Disease, 13(12), 1045.

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Cryosection Immunostaining for Chromatin Modifications

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The cryosection immunostaining was performed as previously described [54 (link)]. The primary antibodies were anti-Wdr5, anti-H3K4me3, anti-H4K16ac, anti-pH3, anti-Bhmt and anti-β-Catenin. DAPI was used to stain nuclear. Images from different samples in each experiment were taken under the same voltage for respective laser channel by a confocal microscope (Olympus BX61WI). The percentage of pH3 positive cells in each sample was calculated as the number of pH3 positive cells divided by total cell number in different organs from continuous cryosections.
The information of all antibodies and the numbers of different cells from each sample were provided in the Supplementary Tables S2, 3 and 7, respectively.
Hematoxylin-Eosin (H&E) staining was performed as previously described [62 (link)] . Images were captured under an Olympus BX53 microscope with a camera from Qimaging MicroPublisher 5.0 RTV.

Zhang Z., Yang C., Wang Z., Guo L., Xu Y., Gao C., Sun Y., Zhang Z., Peng J., Hu M., Jan Lo L., Ma Z, & Chen J. (2023). Wdr5-mediated H3K4me3 coordinately regulates cell differentiation, proliferation termination, and survival in digestive organogenesis. Cell Death Discovery, 9, 227.

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In Vivo Nanoparticle and EV Tracking

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Mice (BALB/c-nu or C57BL/6, 6–8 weeks old, male) were anesthetized by the intraperitoneal injection of 3% chloral hydrate and immobilized in the custom-made stereotactic apparatus under the objective. Saline, nanoparticles (NPs) and EVs were mixed with PKH26 linker kits (Sigma-Aldrich) in a ratio of 1:1, and the mixture was immediately injected intravenously into the four different groups: PBS, NPs (Turbo for BALB/c-nu mice and Liposome for C57BL/6 mice), exosomes and Exo-Ts; (n=3 per group). The upright laser scanning microscope (BX61WI, Olympus) attached to a Ti: sapphire pulsed laser system (80 MHz repetition rate, <100 fs pulse width, Spectra Physics) and software (Prairie view 5.4, Bruker) was used to track and measure the distribution of saline, NPs and EVs within the tumour area at different times after injection: 1h, 4h, 8h, and 24h. 20x water immersion (NA, 1.00; WD, 2 mm, Olympus), and 40x water-immersion objectives (NA 0.80, WD; 3.3 mm, Olympus) were chosen for fluorescence imaging in vivo., 890-nm irradiation wavelength was used to excite U87-Luc (or Gl261-Luc) and PKH26 red fluorescence, and emission light was differentiated and collected with 525/50 and 595/500 filters, respectively. The average laser power for imaging was less than 50 mW.

Yang Z., Shi J., Xie J., Wang Y., Sun J., Liu T., Zhao Y., Zhao X., Wang X., Ma Y., Malkoc V., Chiang C., Deng W., Chen Y., Fu Y., Kwak K.J., Fan Y., Kang C., Yin C., Rhee J., Bertani P., Otero J., Lu W., Yun K., Lee A.S., Jiang W., Teng L., Kim B.Y, & Lee L.J. (2019). Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation. Nature biomedical engineering, 4(1), 69-83.

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