Mica supported DOPC monolayers were prepared by aforementioned protocol. In the FRET measurements, the samples contained 0.2 mol% Bodipy-PC and 0.4 mol% Texas Red-DPPE as FRET donor and acceptor, respectively. Then, the samples were dehydrated by the RFS methods or dehydrated at the atmospheric pressure and the room temperature. Then, the intensity of the FRET-donor Idonor was measured at more than thirty different locations (50 × 50 μm2 for each) in the sample with fluorescent microscope BZ-X700 and the average value was shown. The Idonor-value of the wet sample was measured immediately after the preparation of the supported monolayer. The excitation wavelength of 470 nm was applied to Bodipy-PC and the emission were detected at 525 nm using dichroic mirrors OP-87763 (Keyence, Osaka, Japan).
Op 87763
Manufactured by Keyence
Sourced in Japan
The OP-87763 is a high-precision laser displacement sensor. It is designed to measure the displacement of objects with a high degree of accuracy. The sensor utilizes a laser beam to detect changes in the distance to a target object, and it can provide reliable and consistent measurements.
Automatically generated - may contain errors
Lab products found in correlation
Op 87762, by Keyence (7 mentions)
Hoechst 33342, by Thermo Fisher Scientific (6 mentions)
Bz x710, by Keyence (5 mentions)
Bz x viewer software, by Keyence (3 mentions)
Op 87764, by Keyence (2 mentions)
Plan apochromat 40 objective, by Keyence (2 mentions)
Bz x800, by Keyence (2 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (2 mentions)
Bz x analyzer software, by Keyence (2 mentions)
Cls3770, by Keyence (2 mentions)
Op 87765, by Keyence (2 mentions)
Op 87767, by Keyence (2 mentions)
Bz x700, by Keyence (1 mentions)
All in one fluorescence microscope bz x800, by Keyence (1 mentions)
Bz h4xt, by Keyence (1 mentions)
Bz h4xd, by Keyence (1 mentions)
Biodipy 493 503, by Thermo Fisher Scientific (1 mentions)
Inverted fluorescence microscope bzx 710, by Keyence (1 mentions)
Bz analyzer, by Keyence (1 mentions)
Op 87766, by Keyence (1 mentions)
12 protocols using op 87763
1
FRET Analysis of Dehydrated Lipid Monolayers
2
Electroformation of Giant Unilamellar Vesicles
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GUVs were prepared using an electroformation method40 (link). Briefly, appropriate amounts of the lipid solutions were mixed in a glass vial (1 mg/mL), to which small amounts of fluorescent lipid analogs (0.2 mol% of total lipids) were added. The 10-μL aliquot of the lipid solution was spread on the surface of electrodes, platinum wires (100 μm diameter), and dried under vacuum for more than 12 h. Then parallel aligned electrodes were put into Milli-Q water sandwiched between two cover glasses (24 mm × 60 mm, 0.12–0.17 mm thickness) using an open-square-shaped rubber spacer. This chamber was fixed on a temperature-controlled aluminum block kept at 70 °C (Sahara 310, Rocker Scientific Co., Ltd., Taipei, Taiwan). The sample was incubated for 60 min, applying a low frequency alternating current (AC) field (sinusoidal wave function, 10 Vpp, 10 Hz), by a function generator (20 MHz function/arbitrary waveform function generator, Agilent, Santa Clara CA). The GUVs were then cooled to 25 °C. Fluorescent observation was carried out using the BZ-X700 (Keyence, Osaka, Japan) with an air objective lens (Plan Apoλ, 60 ×, N.A. 0.95, Nikon, Tokyo, Japan). The excitation (470 nm) and detection (525 nm) wavelengths were selected by dichroic mirrors, OP-87763 (Keyence, Osaka, Japan).
3
Imaging Fungal Infection Dynamics
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Inoculated coleoptile was placed onto a glass-bottom culture dish (35 mm in diameter). To avoid drying out, 1 mM CaCl2 was supplied under the coleoptile. To maintain high humidity inside, small pieces of moistened Kimwipe paper were placed around the coleoptile and the lid of the dish was sealed with Parafilm (Figure 5 a). Images of infection behavior were taken under a BZ-X800 all-in-one fluorescence microscope (Keyence, Osaka, Japan). Real-time monitoring was automatically performed by capturing fluorescence images of 17 Z-stacks every 30 min for 48 h using the long-term time-lapse modules (BZ-H4XD and BZ-H4XT, Keyence). A time-lapse movie was generated with BZ-X800 Analyzer (BZ-H4A, Keyence). Green fluorescence of F. graminearum expressing GFP was detected using the GFP filter (OP-87763, Keyence), and red fluorescence of FM4-64 staining the plasma membrane of epidermal cells was detected using the TRITC filter (OP-87764, Keyence).
4
Visualizing Osmotic Stress Response
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Transgenic seedlings (4-day old) grown on an MS agar plate were transferred to a plate supplemented with 600 mM sorbitol for 2 d (osmotic-shock stress). Nuclei in the seedlings were stained in PBS containing 0.5 μg/ml Hoechst 33342 for 20–30 min before fluorescence microscopy. Images were obtained with an all-in-one fluorescence microscope (BZ-X800, Keyence, Osaka, Japan) equipped with an optical sectioning module and a Plan Apochromat 40× objective (NA0.95 and BZ-PA40, respectively, both from Keyence, Tokyo). Green fluorescence was detected with a GFP filter (ex 470/40 nm, em 525/50 nm, dichroic 495 nm; OP-87763, Keyence). Blue fluorescence (Hoechst 33342) was detected with a DAPI filter (ex 360/40 nm, em 460/50 nm, dichroic 400 nm; OP-87762, Keyence).
5
Adipogenesis Quantification via Fluorescence
6
Quantitative Adipogenesis Imaging and Analysis
7
Imaging of Fluorescent Zebrafish Embryos
8
FISH analysis of KDM5D in SCCs
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FISH analysis of the TMAs was performed using a two‐color KDM5D/X chromosome (CenX) probe mix, which was provided by the Molecular Cytogenesis Core at Memorial Sloan Kettering Cancer Center (New York, USA). From the 194 male patients with SCCs, TMAs of 173 tumors were available for FISH analysis. The probe mix consisted of a bacterial artificial chromosome clone containing full‐length KDM5D (Yq11) (RP11‐188C1 and RP11‐204P21; labeled with red dUTP) and a centromeric repeat plasmid specific to CenX (pSV2x5; labeled with green dUTP). Probe labeling, tissue processing, hybridization, post‐hybridization washing, and fluorescence detection were performed according to standard laboratory procedures [15 (link)]. Images were obtained with an all‐in‐one fluorescence microscope (BZ‐X800; KEYENCE, Osaka, Japan) equipped with a Plan Apochromat ×40 objective (NA0.95, BZ PA40; KEYENCE). Red fluorescence was detected using a TexasRed filter (ex: 560/40 nm, em: 630/75 nm, dichroic: 585 nm, OP‐87765; KEYENCE) and green fluorescence was detected using a GFP filter (ex: 470/40 nm, em: 525/50 nm, dichroic: 495 nm, OP‐87763; KEYENCE). A minimum of 50–300 nuclei per case were evaluated. Copy number loss of KDM5D was defined as cases in which more than 90% of tumor cells showed absence of a red (KDM5D) signal, as previously described [15 (link)].
9
Quantitative Adipogenesis Imaging and Analysis
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Adipogenesis was assessed by staining with Bodipy 493/503 (Invitrogen catalog no. D3922) for lipid accumulation and Hoechst 33342 (Thermo Fisher catalog no. 62249) for nuclei32 . Briefly, cells were differentiated in 384 well tissue culture plates (Sigma-Aldrich catalog no. CLS3770), fixed with 4% paraformaldehyde, stained, and imaged on a Keyence inverted microscope (BZX-710) using Keyence BZ-X Viewer Software (1.3.1.1) with the following filters: DAPI (ex, 360/40 nm; em, 460/50 nm; Keyence, OP-87762) and GFP (ex, 470/40 nm; em, 525/50 nm; Keyence, OP-87763) filters. Images were acquired at 20x in a 7×7 tiled grid and stitched to capture the entirety of each well. Tiling and stitching were performed with Keyence BZ-X Analyzer software (1.3.0.3). Image quantification was performed automatically in ImageJ (version 1.52E) using a macro which: 1) Split images into component channels, 2a) for the nuclei channel applied a 3-Sigma Gaussian blur, performed thresholding to identify signal above background, performed watershed to segmentation, and counted the number of nuclei, 2b) for the lipid channel applied a 2-Sigma Gaussian blurm performed thresholding to identify signal above background, and counted the area (#of pixels) with signal above threshold. The amount of adipogenesis was calculated as Lipid Area/#nuclei.
10
Fluorescence Microscopy for Subcellular Analysis
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The cells were visualized using a fluorescence microscope (BZ-X710; Keyence Co., Osaka, Japan). To visualize the VP, a filter cube (OP-87767; Keyence Co., Osaka, Japan) was used with relevant excitation (405BP20) and fluorescence (RPE630LP) filters. To visualize the MitoBright Green and singlet oxygen, the BZ-X filters GFP (OP-87763; Keyence Co., Osaka, Japan) were used. To visualize the Hoechst staining, the BZ-X filters DAPI (OP-87762; Keyence Co., Osaka, Japan) were used. The software BZ-analyzer (Ver.1.3.1.1., Keyence Co., Osaka, Japan) was used to merge, reduce noise, and enhance signal intensity.
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