1.4 na objective

Manufactured by Nikon

The 60× 1.4 NA objective is a high-precision optical lens designed for use in laboratory equipment. It features a magnification of 60× and a numerical aperture (NA) of 1.4, which enables it to gather a large amount of light and provide high-resolution imaging. This objective is suitable for a variety of microscopy applications that require detailed, high-quality observations.

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Lab products found in correlation

A1r laser scanning confocal microscope, by Nikon (2 mentions) 90i upright microscope, by Nikon (2 mentions) Desflurane, by Baxter (1 mentions) Csu w1 sora, by Yokogawa (1 mentions) 2 3 butanedione monoxime, by Merck Group (1 mentions) Specrax led illuminator, by Lumencor (1 mentions) 582 669 di01 and di01 r442 514 561 dichroic mirrors, by IDEX Corporation (1 mentions) Hamamatsu flash 4.0 scmos camera, by Sutter Instruments (1 mentions) Ti e microscope, by Nikon (1 mentions) Orca flash 4 camera, by Hamamatsu Photonics (1 mentions) Ti e inverted microscope, by Nikon (1 mentions) Nis element, by Nikon (1 mentions)

Close spelling variants of this product

Plan Apo 1.4 NA objective 63 × Plan Apochromat 1.4 NA objective Plan Apo 1.4 NA oil objective 60 × 1.4 NA oil immersion objective Apo-Plan 60 × 1.4 NA oil objective VC Plan Apo 60X/1.4 NA oil objective 100x 1.4 NA oil DIC Plan-Apochromat VC objective Nikon plan Apo 100×/NA 1.4 objective Plan Apochromat VC PFS 1.4 NA oil-immersion objective PLAN APO 100X 1.4 NA DIC H Nikon objective

7 protocols using 1.4 na objective

Imaging Network Dynamics with Laser Microscopy

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Networks are imaged using a Nikon A1R laser scanning confocal microscope with a 60× 1.4 NA objective (Nikon). Actin filaments are imaged using a 561 nm laser with 561 nm excitation and 595 nm emission filters. Microtubules are imaged using a 488 nm laser with 488 nm excitation and 525 nm emission filters. Myosin II motor activity is controlled by deactivating blebbistatin with the same 488 nm excitation used to image microtubules (continuous 360 ms pulses of 488 nm light over the duration of each experiment). While blebbistatin deactivation is typically achieved using 405 nm light,^{17 (link)} previous studies have shown successful photoinactivation of blebbistatin using 488 nm light.^{37 (link),55 (link)} 256 × 256 pixel images (212 μm × 212 μm) are taken in the middle of the ~70 μm thick chamber for 45 min at 2.78 fps. Examples of time-series are shown in Movie S1 (ESI†), and representative images are shown in Fig. 1. Experiments are performed on 3–5 different replicates.

Lee G., Leech G., Lwin P., Michel J., Currie C., Rust M.J., Ross J.L., McGorty R.J., Das M, & Robertson-Anderson R.M. (2021). Active cytoskeletal composites display emergent tunable contractility and restructuring. Soft matter, 17(47), 10765-10776.

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Imaging GFP-Labeled Organelles in Larval Axons

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Time-lapse imaging of GFP-loaded organelles in axons was done with living third instar larvae 4–5 d after egg lay. A larva was placed in an imaging chamber modified from Fuger et al. (2007 (link)), anesthetized by injecting 50 ul of chilled Desflurane (Baxter) into the chamber, and sealed. For each animal, segmental nerve 7 or 8, in segments A4–A5, was imaged through the ventral body wall with the spinning disk microscope using a 60× 1.4 Na objective (Nikon). Images of DCVs and mitochondria were collected at 2 frames/s and 1 frame/s, respectively, for at least 500 frames. After imaging, larvae were returned to normal culture medium. Data were analyzed only from larvae that survived the procedure and recovered crawling mobility. Although larvae can survive in the anesthesia chamber for hours, imaging was restricted to the initial 30 min to reduce the potential for transport variation from physiological stress.

Lim A., Rechtsteiner A, & Saxton W.M. (2017). Two kinesins drive anterograde neuropeptide transport. Molecular Biology of the Cell, 28(24), 3542-3553.

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Confocal Imaging of Fluorescent Signals

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The confocal imaging was performed with a A1-R confocal laser scanning microscope (Nikon), equipped with a Nikon 60×, 1.4 NA objective, and with a 488 and 561 nm laser lines to excite FITC (green) and TRITC (red) fluorescence signals. The 3D images were processed by stacking up 20–25 consecutive confocal images with surface shaded reconstruction. No deconvolution was applied to the images.

Sabatelli P., Castagnaro S., Tagliavini F., Chrisam M., Sardone F., Demay L., Richard P., Santi S., Maraldi N.M., Merlini L., Sandri M, & Bonaldo P. (2014). Aggresome–Autophagy Involvement in a Sarcopenic Patient with Rigid Spine Syndrome and a p.C150R Mutation in FHL1 Gene. Frontiers in Aging Neuroscience, 6, 215.

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Visualizing GABA Receptor Dynamics

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Super-resolution images were captured with a Nikon 60 × 1.4 NA Objective on a Nikon spinning-disk confocal system (Yokogawa CSU-W1 SoRa) based on SoRa mode. Live young adult animals were anesthetized with 30 μg/μl 2,3-Butanedione monoxime (Sigma) and the regions of dorsal cords were excited by a 561 nm laser (50% power, 400 ms exposure time). The maximum intensity of dorsal cord projections of Z-series stacks was obtained by ImageJ. The number of animals with diffusing or punctate GABA_ARs was assessed and analyzed by Chi-square test.

Hao Y., Liu H., Zeng X.T., Wang Y., Zeng W.X., Qian K.Y., Li L., Chi M.X., Gao S., Hu Z, & Tong X.J. (2023). UNC-43/CaMKII-triggered anterograde signals recruit GABAARs to mediate inhibitory synaptic transmission and plasticity at C. elegans NMJs. Nature Communications, 14, 1436.

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Visualizing antibiotic-induced changes in E. coli

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Following the protocol of Nonejuie et al. (2013) (link), overnight E. coli lptD4213 cultures were diluted 1:100 in LB and grown at 30°C for 90 minutes on a roller drum. Each culture was then treated with 5X the MIC and incubated at 30°C with slight agitation. Following antibiotic treatment, cells were stained with 0.5 μM SYTOX Green, 1 μg/mL FM4–64, and 2 μg/mL DAPI for incubated for 10 minutes. Each stained culture was then centrifuged at 3220 rcf. for 40 s and resuspended in 1/10 volume of LB. Cells spotted onto a 1.2% agarose pad in 20% LB medium for imaging. Images were collected on a Nikon 90i upright microscope equipped with a 100X 1.4 N.A. objective (Nikon Instruments Inc., Melville, NY) and a RoleraXR (Photometrics, Tucson, AZ) camera. Microscope control and image acquisition were performed in NIS Elements.

Martin JK I.I., Sheehan J.P., Bratton B.P., Moore G.M., Mateus A., Li S.H., Kim H., Rabinowitz J.D., Typas A., Savitski M.M., Wilson M.Z, & Gitai Z. (2020). A Dual-Mechanism Antibiotic Kills Gram-Negative Bacteria and Avoids Drug Resistance. Cell, 181(7), 1518-1532.e14.

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Immunofluorescence and Live-Cell Imaging Protocol

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Cells for immunofluorescence were grown on glass coverslips and fixed in 3.7% formaldehyde in 200 mM Hepes pH 7.2. Staining for immunofluorescence and digital photography were done as described before (Karanasios et al., 2013 (link)). Live-cell imaging was performed as previously described (Karanasios et al., 2013 (link)) using a Nikon Ti-E-based system. Cells plated onto 60 plastic dishes were transferred onto 22-mm-diameter glass coverslips (BDH) and secured in an imaging chamber with 2 ml of cell medium or starvation medium added as indicated. The assembled imaging chamber was secured onto the microscope stage, and cells were maintained at 37°C using an OKO Labs full enclosure incubation system. The Nikon Ti-E-based system comprised a Nikon Ti-E microscope, 100x 1.4 N.A. objective (Nikon), SpecraX LED illuminator (Lumencor, Beaverton, OR), 410/504/582/669-Di01 and Di01-R442/514/561 dichroic mirrors (Semrock), Hamamatsu Flash 4.0 sCMOS camera, emission filter wheel (Sutter Instruments) and was controlled using Nikon Elements software. Compounds (fluorescent analogue, amino acids, and drugs) were added during imaging by flushing the solution in the imaging chamber with 5 ml of fresh solution containing the indicated additions.

Manifava M., Smith M., Rotondo S., Walker S., Niewczas I., Zoncu R., Clark J, & Ktistakis N.T. (2016). Dynamics of mTORC1 activation in response to amino acids. eLife, 5, e19960.

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Microscopic Visualization of Antibiotic-Treated E. coli

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In dual-treatment experiments (Figures 6A and S6), overnight E. coli lptD4213 cultures were diluted 1:100 and grown to early-mid exponential phase (OD₆₀₀ = 0.4–0.6). Each culture was then diluted 1:10 into fresh LB and treated with the desired concentration of antibiotic for 10 minutes. Following antibiotic treatment, cells were stained with 0.5 μM SYTOX Green, 1 μg/mL FM4–64, and 2 μg/mL DAPI. Each stained culture was then spotted onto a 1.5% agar pad supplemented with casamino acids and glucose in M63 (15 mM (NH₄)₂SO₄, 100 mM KH₂PO₄, 1.7 μM FeSO₄, 0.5% glucose, 0.2% casamino acids, 1 mM MgSO₄). Images were collected on either Nikon 90i upright microscope equipped with a 100X 1.4 N.A. objective (Nikon) or a Nikon Ti-E inverted microscope equipped with a 100X 1.4 NA objective. Both microscopes utilize an Orca Flash4 camera (Hamamatsu, Bridgewater, NJ). Microscope control and image acquisition were performed in NIS Elements (Nikon).

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