Uplanfln objective

Manufactured by Olympus

Sourced in Australia

The UPlanFLN objective is a high-quality optical lens designed for use in Olympus microscopes. It is intended to provide clear and detailed imaging for various microscopy applications.

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UPlanFLN (47 citations) UPlanFLN 40×/1.30 oil objective (4 citations) UPlanFLN 60× objective lens (3 citations) UPlanFLN 40× immersion oil objective (2 citations) Olympus UIS2 UPlanFLN 100X/1.30 OilPh3 objective (2 citations)

13 protocols using uplanfln objective

Histological Analysis of Muscle Tissue

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Evans blue dye (EBD) (1%, Sigma-Aldrich) was intraperitoneally injected [29 (link)]. Twenty hours after injection, muscle tissues were harvested. Microscopic images were captured by a DP-70 digital microscope camera attached to BX51 microscope with 4 x and 10 x UPlanFLN objectives (all from Olympus). Photoshop CS2 (Adobe Systems) was used for image processing. Freshly dissected tissues were frozen and sectioned at 8 μm thickness by cryostat (Leica CM1900). Sections were stained with hematoxylin and eosin (HE) for quantification of fibers with centrally located nuclei (CLN) and fiber diameters. Sections were also stained with Alizarin red (Sigma-Aldrich) and Sirius red (Sigma-Aldrich) for the assessment of calcification and fibrosis, respectively. The quantifications were done using the entire section area. Quantitative analysis was performed by the ImageJ software from the NIH.

Shimizu-Motohashi Y., Asakura Y., Motohashi N., Belur N.R., Baumrucker M.G, & Asakura A. (2015). Pregnancy-Induced Amelioration of Muscular Dystrophy Phenotype in mdx Mice via Muscle Membrane Stabilization Effect of Glucocorticoid. PLoS ONE, 10(3), e0120325.

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High-Resolution Imaging of Vaginal Tissue

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Images of immunostaining were acquired with Olympus BX51 upright fluorescence microscope equipped with Olympus UPlanFLN objectives (x40, NA = 0.75; x60, NA = 0.9) and Olympus DP72 CCD (charge-coupled device) color camera using cellSens software (Olympus), or Zeiss AxioImager A1 upright fluorescence microscope equipped with AxioCam MRc camera and Zeiss EC Plan-NEOFLUAR objectives (x20, NA = 0.5; x40, NA = 0.75) using ZEN software or Olympus Fluoview FV10i upright confocal laser scanning microscope fitted with 60x oil-immersion objective (NA = 1.35) (Olympus), using FV10i-SW software. Whole mount imaging of 100um thick slices of X-gal stained vaginae was done on SMZ25 stereoscope equipped with Nikon DS-Fi2 camera and Nikon P2-SHR Plan Apo 2X objective (NA = 0.312). Images consisted of 15-30 stacks with variable z-spacing and were captured and merged using NIS-Elements software. All images were captured at room temperature.

Ali A., Syed S.M., Jamaluddin M.F.B., Colino-Sanguino Y., Gallego-Ortega D, & Tanwar P.S. (2020). Cell Lineage Tracing Identifies Hormone-Regulated and Wnt-Responsive Vaginal Epithelial Stem Cells. Cell reports, 30(5).

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NBT Staining for Superoxide Detection in E. festucae

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Superoxide radicals in E. festucae in axenic culture were stained using NBT (Sigma-Aldrich) as described by Tanaka et al. (2006 (link)). Three replicates for each strain were included in the analysis and the experiment was repeated twice. Strains were grown on PDA for 1 week at 22°C under 8 h light and 16 h dark, and mycelia stained for 5 h at 22°C in continuous light by incubation in 20 μL of 0.05% (w/v) NBT dissolved in 0.05 M sodium phosphate pH 7.5. The reaction was stopped by removing the stain and adding 40 μL of absolute ethanol. The ethanol was removed after 5 min, and mycelia were analyzed at 400X magnification under bright field illumination using an Olympus BX50 compound microscope with a 40X UPLANFLN objective and a 0.75 numerical aperture. Images were taken using an Olympus Colorview III camera with AnalySIS^B image processing software.

Voisey C.R., Christensen M.T., Johnson L.J., Forester N.T., Gagic M., Bryan G.T., Simpson W.R., Fleetwood D.J., Card S.D., Koolaard J.P., Maclean P.H, & Johnson R.D. (2016). cAMP Signaling Regulates Synchronised Growth of Symbiotic Epichloë Fungi with the Host Grass Lolium perenne. Frontiers in Plant Science, 7, 1546.

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Histopathological Assessment of Mouse Tissues

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Mice were euthanized with CO₂. The kidneys, liver, and lungs were removed and fixed for 48–72 h in 10% neutral buffered formalin. One kidney was trimmed longitudinally and the contralateral kidney was trimmed transversally in the mid portion. For the liver, one sample from the left lobe and two samples from the median lobe were trimmed. The lungs were processed whole without trimming. Tissues were processed to paraffin blocks, and 4-μm sections were stained with haemotoxylin and eosin (H&E) or Periodic Acid Schiff (kidney only). Kidney, liver, and lung tissue sections were assessed by a veterinary pathologist (F.M.) blinded to the genotype of the mice in the different cohorts. Ordinal semiquantitative scoring criteria used for the assessment of lesions in the kidneys, liver, and lungs were defined according to guidelines and principles of histopathologic scoring in research (Gibson-Corley et al, 2013 (link)) and are detailed below. The photomicrographs presented in the article were captured using an Olympus BX53 with microscope UPlanFL N objective. The ocular lens was WHN10×-H/22 and the camera used was an Olympus SC100. For preparation of the figures, images were obtained and processed using an Olympus CellSens Standard and resized using Adobe Photoshop.

Nanda S.K., Petrova T., Marchesi F., Gierlinski M., Razsolkov M., Lee K.L., Wright S.W., Rao V.R., Cohen P, & Arthur J.S. (2019). Distinct signals and immune cells drive liver pathology and glomerulonephritis in ABIN1[D485N] mice. Life Science Alliance, 2(6), e201900533.

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Fluorescence Lifetime Imaging of GFP and DNA

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Fluorescence lifetime measurements were made using a Becker & Hickl FLIM system attached to an Olympus IX81 microscope, equipped with a 40x (1.3 N.A.) oil UPlan FLN objective. Fluorescence excitation was produced with a pulsed 488 nm laser, operating at 80 MHz. Analysis was performed using SPCImage software. Samples contained either GFP WT (superfolder GFP 29, 32 ) or 5'--Cy3 ssDNA (12mer), and were prepared as for partitioning experiments, described below.

Nott T.J., Craggs T.D, & Baldwin A.J. (2016). Membraneless organelles can melt nucleic acid duplexes and act as biomolecular filters. Nature chemistry, 8(6).

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Quantitative Imaging of Hepatocyte Proliferation and Apoptosis

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Concerning HE, BrdU and PHH3 labelling, images were taken using a Nikon Statif Eclipse E600 microscope with x10 and x20 magnification, 1.4–0.7 NA PL-APO objectives, a DXM1200 cooled CCD camera (Nikon), and ACT-1 (version 2.63; Universal Imaging). For cleaved-caspase 3 labelling, images were taken using an Olympus BX63F, at 4x magnification Uplan FLN objective, an Olympus DP73 camera and Metamorph software. Necrotic area were quantified by morphometric analysis using an open-source ImageJ software in 5 fields at x10 magnification. For BrdU/PHH3 staining, 4000 hepatocytes (for each liver sample analyzed) were counted; at least 10 areas of 33,500 μm² were analyzed. Cleaved-caspase 3 immunostaining was quantified by color segmentation using an open-source ImageJ software in 5 fields at 4x magnification. Adobe Photoshop CS (Adobe Systems Software) was used for figure construction.

Fortier M., Cadoux M., Boussetta N., Pham S., Donné R., Couty J.P., Desdouets C, & Celton-Morizur S. (2019). Hepatospecific ablation of p38α MAPK governs liver regeneration through modulation of inflammatory response to CCl4-induced acute injury. Scientific Reports, 9, 14614.

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Membrane Imaging and Temperature Control

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Membranes were imaged by epifluorescent microscopy on an Olympus IX71 inverted microscope at 100Â magnification with an Olympus TIRF UIS2 UAPON objective and by confocal microscopy on an Olympus IX81 inverted microscope at 100Â magnification with an Olympus UPlanFl N objective. Temperatures were controlled using a Physitemp TS-4 SPD Controller. When samples were heated, the temperature reading was calibrated to a measured temperature calibration curve which we constructed by inserting a thermocouple into one of our sample chambers and measuring the sample temperature as a function of the controller's temperature reading. Data analysis was done in ImageJ and Matlab.

Shindell O., Mica N., Ritzer M, & Gordon V.D. (2015). Specific adhesion of membranes simultaneously supports dual heterogeneities in lipids and proteins. Physical chemistry chemical physics : PCCP, 17(24).

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DAF-FM Imaging of Nitric Oxide

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As previously described^{29 (link)} DAF-FM imaging was performed according to standard methods using the 488 nm argon laser line of a Fluoview FV1000 laser scanning confocal system and IX-81 microscope (10×, 0.3 NA UPlanFLN objective; Olympus). Cells were loaded with DAF-FM by incubation in DPBS containing 10 μM DAF-FM diacetate in PBS containing 5 μM carboxy PTIO, a cell permeant NO scavenger (on the apical side only). After 30 min, cultures were copiously washed (at least 5 times) with PBS to remove all traces of unloaded DAF-FM. Cultures were then incubated for ~15 min to allow for dye retention before imaging was performed. DAF-FM fluorescence images were acquired at 5 second intervals. Because the magnitudes of DAF-FM fluorescence changes are used to approximate NO production, care was taken to follow the loading protocol strictly to normalize dye loading and microscope/software settings were identical for each experiment.

, & Cohen N.A. (2016). The Genetics of the Bitter Taste Receptor T2R38 in Upper Airway Innate Immunity and Implications for Chronic Rhinosinusitis. The Laryngoscope, 127(1), 44-51.

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Quantifying Mitotic Spindle Dynamics

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FDAPA analysis and images used for quantifying astral length, inner spindle intensity, spindle orientation, and TACC3 and MCAK spindle localization were acquired using a Marianas (Intelligent Imaging Innovations) spinning-disk confocal system based on a microscope (Axio Observer Z1; ZEISS) equipped with a camera (ORCA-Flash 4.0; Hamamatsu Photonics). Images were taken using a 63× 1.4 NA Apochromat objective (ZEISS). Images for TACC spindle localization in U2OS cells were taken using a 100× 1.4 NA Plan-Apochromat objective (ZEISS). The images were z-projected using Slidebook software 5.5 (Intelligent Imaging Innovations). All other images were acquired using a DeltaVision imaging system (GE Healthcare) equipped with an sCMOS camera (PCO Edge 5.5). Images were taken using a 60× 1.42 NA PlanApo-N objective (Olympus) at room temperature. Serial z stacks of 0.2-µm thickness were obtained and deconvolved using SoftWoRx software 6.1.l. For live-cell imaging, media was changed to CO₂-independent media (Gibco) 12 h before imaging. Live-cell image sequences were acquired at 1-min intervals for 12 h in 2-µm serial z sections using a 40× 1.42 NA UPlanFL-N objective (Olympus) at 37°C.

Bendre S., Rondelet A., Hall C., Schmidt N., Lin Y.C., Brouhard G.J, & Bird A.W. (2016). GTSE1 tunes microtubule stability for chromosome alignment and segregation by inhibiting the microtubule depolymerase MCAK. The Journal of Cell Biology, 215(5), 631-647.

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Fluo-4 Imaging and Live/Dead Staining of ALIs

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Fluo-4 loading and imaging were done exactly as previously described (8 (link), 14 (link), 92 (link)). Cells were loaded with 10 μM fluo-4-AM (PBS + 0.1% pluronic F127; apical side only) for ~120 min at room temperature, followed by 3 washes with DPBS and 15–20 min incubation to allow cells to recover. Images were captured at 3 or 5 sec intervals using the 488 nM laser line of an Olympus Fluoview confocal system attached to an Olympus IX81 microscope (10×, 0.3 NA UPlanFLN objective; Olympus). No gain, offset or gamma alterations were used. Normalization of Fluo-4 fluorescence changes was made after subtraction of background, approximated for each experiment by measuring unloaded ALIs. Baseline fluorescence (F_o) was determined by averaging the first 10 frames of each experiment.
Live/Dead staining of ALIs was performed using Syto9/PI mix of a BacLight^® live/dead kit. MRSA-M2 overnight cultures were centrifuged, normalized to OD 0.01, and incubated with human ALI cultures for 6 hours at 37° C. Afterward, ALIs were washed copiously with PBS and stained using the two dyes at concentrations indicated in the manufacturers instructions and imaged immediately.

Lee R.J., Hariri B.M., McMahon D.B., Chen B., Doghramjii L., Adappa N.D., Palmer J.N., Kennedy D.W., Jiang P., Margolskee R.F, & Cohen N.A. (2017). Bacterial D-Amino Acids Suppress Sinonasal Innate Immunity Through Sweet Taste Receptors in Solitary Chemosensory Cells. Science signaling, 10(495), eaam7703.

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