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Oil objective

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The 60× oil objective is a high-magnification lens designed for use with microscopes. It provides a 60× magnification and is intended for use with oil immersion techniques to achieve high-resolution imaging of small samples.

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Lab products found in correlation

Rat tail collagen 1, by Thermo Fisher Scientific (3 mentions) C8484 camera, by Hamamatsu Photonics (3 mentions) Hci image software, by Hamamatsu Photonics (3 mentions) Poly d lysine, by Merck Group (3 mentions) Ix71 microscope, by Olympus (3 mentions) Glass bottom dishes, by MatTek (2 mentions) Metamorph 7.5 imaging software, by Molecular Devices (2 mentions) Fv1000 confocal laser scanning microscope, by Olympus (2 mentions) Axiovert s100 microscope, by Eppendorf (2 mentions) Spin x filter, by Corning (2 mentions) Ix70 imaging system, by Olympus (2 mentions) Femtojet microinjector, by Eppendorf (2 mentions) Femtotip, by Eppendorf (2 mentions) Deltavision elite, by GE Healthcare (1 mentions) Em ccd camera, by Hamamatsu Photonics (1 mentions) Tmr ligand, by Promega (1 mentions) D2 phaser xrd, by Bruker (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Dspe peg2000 biotin, by Avanti Polar Lipids (1 mentions) Ix71 inverted microscope, by Olympus (1 mentions)

37 protocols using oil objective

1

Mitochondrial Morphology Quantification

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MEF or HeLa cells transiently transfected with mtGFP or MEFmtGFP cells were seeded at a density of 100,000 cells/well on glass-bottom dishes (MatTek) coated with poly-D-lysine (Sigma) or rat tail collagen 1 (GIBCO). Cells were then treated as indicated and images were recorded with an Olympus IX71 microscope with 60x oil objective (Olympus), a Hamamatsu C8484 camera (Hamamatsu Photonics), and HCI image software (Hamamatsu Photonics). Quantification was performed by blinding the images and then scoring cells based on the presence of primarily fragmented, tubular, or elongated mitochondria, as before (Lebeau et al., 2018 (link)). At least three different researchers scored each set of images and these scores were averaged for each individual experiment and all quantifications shown were performed for at least 3 independent experiments quantifying a total of >60 cells/condition across all experiments. The data were then analyzed in PRISM (GraphPad, San Diego, CA) and plotted on a stacked bar plot to show the average morphology and standard error of the mean across all experiments. Statistical comparisons were performed using a 2-way ANOVA in PRISM, comparing the relative amounts of fragmented, tubular, or elongated mitochondria across different conditions.
Perea V., Baron K.R., Dolina V., Aviles G., Rosarda J.D., Guo X., Kampmann M, & Wiseman R.L. (2023). Pharmacologic Activation of a Compensatory Integrated Stress Response Kinase Promotes Mitochondrial Remodeling in PERK-deficient Cells. bioRxiv.
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2

Visualizing GABA-Ergic Neurons in C. elegans

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Strains with GFP marked gamma-aminobutyric acid (GABA)-ergic motor neurons were generated by crossing hse-5(tm472) with CK423 and the reporter strain EG1285 oxIs12[Punc-47::GFP + lin-15(+)]. Animals were grown to the indicated stage at 20°C before scoring. Living animals were immobilized on a 2% agarose pad with 0.01% sodium azide, and intact GABA-ergic neurons (L4 or day 1 adult) or axon abnormalities (L4 stage) were scored under fluorescence microscopy on a DeltaVision Elite (GE) imaging system using an Olympus 60x oil objective. Statistical significance was analyzed using Prism statistical software.
Liachko N.F., Saxton A.D., McMillan P.J., Strovas T.J., Keene C.D., Bird T.D, & Kraemer B.C. (2019). Genome wide analysis reveals heparan sulfate epimerase modulates TDP-43 proteinopathy. PLoS Genetics, 15(12), e1008526.
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3

Imaging Halo-Tagged T Cell Activation

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Halo-LZTFL1 or Halo-LZSTOP expressing Jurkat cells were stained with TMR ligand (Promega, Madison, WI) at 37°C and 5% CO2 for 40 min and washed with complete media for 4 times followed by incubation at 37°C and 5% CO2 for 1 h. After washing cells once with complete media, cells were then incubated with 5 μg/ml SEE superantigen-loaded Raji B cells prestained with Hoechst as APCs on poly-L-lysine coated glass-bottom dishes. Cells were maintained at 37°C and 5% CO2 during the entire period of imaging. Halo-tagged T cell images were acquired by fast wide field imaging using an Olympus TIRF 3 system with a 60x oil objective (1.49 NA), under non-TIRF mode. The wide field illumination was achieved by focusing the laser beams into the back focal plane of the objective. Samples labeled with HaloTag and Hoechst for nucleus were excited with 561 nm and 405 nm lasers respectively and the fluorescence signals were extracted with filters LF561-A-OMF for red (Semrock, Rochester, New York) and LF405-A-OMF for blue then recorded with an EMCCD camera (Hamamatsu, New Jersey, USA). Images of the two color channels were acquired in a time course of 10-second interval without interruption. The Acquired images were first processed to remove background and then combined into a movie with home Matlab codes on a Matlab programming environment (Mathwork, MA, USA).
Jiang H., Promchan K., Lin B.R., Lockett S., Chen D., Marshall H., Badralmaa Y, & Natarajan V. (2015). LZTFL1 Upregulated by All-trans Retinoic Acid during CD4+ T Cell Activation Enhances IL-5 Production. Journal of immunology (Baltimore, Md. : 1950), 196(3), 1081-1090.
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4

Correlated DF/FL Imaging of Lipid-Wrapped Nanocomposites

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For the correlated DF/FL imaging of L-NR-NCs with a membrane dye to test successful lipid wrapping (Fig. S2B), 3 mol% DSPE-PEG(2000)-Biotin (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000](ammonium salt), Avanti Polar Lipids) was included in the preparation, and the DPPC was reduced to 52 mol% to keep the total lipid amount constant. The remaining synthesis procedures are identical to described above. 17 μL 1 mg/mL solution of a streptavidin Alexa Fluor 594 (ThermoFisher) dye in 0.5x Phosphate Buffered Saline (PBS) was then added to 50 μL of the nanocomposite suspension and incubated in dark at RT for 2 hours. After centrifuging and washing twice with 0.5x PBS buffer, the samples were observed under an Olympus IX71 Inverted Microscope for DF and FL imaging with an Olympus 60x oil objective (NA = 0.65–1.25). Images were analyzed by ImageJ, and Manders coefficients were calculated with the JACoP ImageJ plugin.76 (link) X-Ray Diffraction spectra were acquired on a Bruker D2 Phaser XRD with a 2 mm receiving slit and a 2.5° Söller slit.
An X., Kays J.C., Lightcap I.V., Ouyang T., Dennis A.M, & Reinhard B.M. (2022). Wavelength-Dependent Bifunctional Plasmonic Photocatalysis in Au/Chalcopyrite Hybrid Nanostructures. ACS nano, 16(4), 6813-6824.
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5

Mitochondrial Morphology Quantification

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HeLa cells transfected with mt GFP or MEF mtGFP cells were seeded at a density of 100,000 cells/well on glassbottom dishes (MatTek) coated with poly-D-lysine (Sigma) or rat tail collagen 1 (GIBCO). Cells were then treated as indicated and images were recorded with an Olympus IX71 microscope with 60x oil objective (Olympus), a Hamamatsu C8484 camera (Hamamatsu Photonics), and HCI image software (Hamamatsu Photonics). At least 20 cells were imaged per condition for each experiment for quantification. Quantification was performed by blinding the images and then scoring cells based on the presence of primarily fragmented, tubular, or elongated mitochondria, as before 38 (link) . Three different researchers scored each set of images and these scores were averaged for each individual experiment. All quantifications shown were performed for at least 3 independent experiments, where averages in morphology quantified from each individual experiment were then combined. The data were then prepared in PRISM (GraphPad, San Diego, CA) and plotted on a stacked bar plot to show the average morphology and standard error of the mean across all experiments.
Statistical comparisons were performed using a 2-way ANOVA in PRISM, comparing the relative amounts of fragmented, tubular, or elongated mitochondria across different conditions.
Perea V., Cole C.M., Lebeau J., Dolina V., Baron K.R., Madhavan A., Kelly J.W., Grotjahn D.A., & Wiseman R.L. (2022). PERK Signaling Promotes Mitochondrial Elongation By Remodeling Membrane Phosphatidic Acid.
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6

Immunofluorescence Staining of mCCD Cells

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500,000 mCCDcl1 cells were seeded onto glass coverslips and after 48 h fixed with 4% paraformaldehyde for 10 min. After washing with PBS, cells were blocked and permeabilized with PBS containing 0.3% Triton X and 3% BSA for 30 min, followed by incubation of primary antibodies in the same buffer for 1 h. The cells were then washed with PBS and incubated with Alexa Fluor 488 anti-rabbit and 647 anti-mouse secondary antibodies (#A-21206 and #A-21449; Invitrogen) for a further hour in PBS. Finally, coverslips were mounted onto slides using Fluoromount-G with DAPI mounting medium (#00-4959-52; Invitrogen). Images were acquired as a z-stack using an Olympus confocal microscope with a 63x oil objective, and all microscope parameters were kept constant from sample to sample.
Rickman O.J., Guignard E., Chabanon T., Bertoldi G., Auberson M, & Hummler E. (2024). Tmprss2 maintains epithelial barrier integrity and transepithelial sodium transport. Life Science Alliance, 7(3), e202302304.
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7

Immunofluorescence Analysis of Embryoid Bodies

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16-µm cryosections of EBs were fixed with 4% PFA (Interchim) during 5 min, washed with PBS, and then incubated 5 min with PBS-Triton X-100 (0.1% v/v). Nonspecific sites were blocked with 5% (w/v) bovine serum albumin (BSA) diluted in PBS during 1 hour and incubated with primary antibodies overnight at 4 °C. Anti-Brachyury T (Abcam, 1:200) and anti-Nkx2.5 (Santa Cruz Technologies, 1:100) primary antibodies were used. The binding of primary antibodies was detected by incubation for 3 h with Alexa Fluor-conjugated anti-rabbit IgG (Cell Signaling, 1:1000). Finally, cells were washed in PBS and mounted with Prolong®Diamond Antifade Mounting Medium with DAPI (Life Technology) for nuclear staining. Cells were analyzed with an Olympus JX81/BX61 device/Yokogawa CSU device spinning-disk microscope (Andor Technology), equipped with a 63X oil objective (Olympus).
Du V., Luciani N., Richard S., Mary G., Gay C., Mazuel F., Reffay M., Menasché P., Agbulut O, & Wilhelm C. (2017). A 3D magnetic tissue stretcher for remote mechanical control of embryonic stem cell differentiation. Nature Communications, 8, 400.
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8

Live-cell Microscopy Imaging Protocol

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Live-cell imaging experiments were mostly performed using an IX-71 (Olympus) microscope with a 63x oil objective (Olympus) (with additional 1.6x optical zoom) and a Cool-SNAP HQ charge-coupled device camera (Photometrics) or an ORCA-Flash4.0 LT Digital CMOS camera (Hamamatsu). Time-lapse imaging experiments were performed with either 2-min or 5-min intervals, and between 3 and 11 0.5-µm z-stacks were taken at each time point. Images shown were mostly maximum intensity projection. Micrographs were taken and analyzed using MetaMorph 7.5 imaging software (Molecular Devices).
Phua S.C., Chiba S., Suzuki M., Su E., Roberson E.C., Pusapati G.V., Setou M., Rohatgi R., Reiter J.F., Ikegami K, & Inoue T. (2017). Dynamic remodeling of membrane composition drives cell cycle through primary cilia excision. Cell, 168(1-2), 264-279.e15.
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9

Fluorescence Microscopy of Apoptosis

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Cells were observed with an Olympus JX81/BX61 device/Yokogawa CSU device spinning-disk microscope (Andor Technology plc, Northern Ireland), equipped with a 63X oil objective (Olympus). Cells were fixed with 4% PFA at room temperature for 15 min. For m-THPC and DAPI, the excitation wavelength was 405 nm, and fluorescence emission was collected with filters at 685 and 445 nm, respectively. Annexin-A5-FITC excitation was carried out at 488 nm and fluorescence emission was collected using a filter at 525 nm. Image J software was used to process the images.
Aubertin K., Silva A.K., Luciani N., Espinosa A., Djemat A., Charue D., Gallet F., Blanc-Brude O, & Wilhelm C. (2016). Massive release of extracellular vesicles from cancer cells after photodynamic treatment or chemotherapy. Scientific Reports, 6, 35376.
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10

Visualizing Mitochondria in C. elegans

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Whole animal images were acquired on an Olympus MVX10 Fluorescence MacroZoom dissecting microscope, using a 540-580 nm excitation filter and 590-670 nm emission filter. Mitochondrial images were taken on a FV1000 Olympus laser scanning confocal microscope using a 100x oil objective (Olympus, N.A. 1.40). Diode laser illumination was 561 nm for red fluorescent transgene and 488 nm for fluorescence of MitoTracker Green FM. Animals were stained with 12 μM MitoTracker Green FM for 20 hr where indicated. MitoTracker stain was dissolved in DMSO, diluted in M9 media (22 mM KH2PO4, 42 mM Na2HPO4, 86 mM NaCl, 1 mM MgSO4, pH 7) and added to OP50 food on culture plates (DMSO <0.02% final concentration) and allowed to dry (Dingley et al. 2010 (link)). Profile plots of pixel intensity were generated using ImageJ software.
Philip N.S., Escobedo F., Bahr L.L., Berry B.J, & Wojtovich A.P. (2019). Mos1 Element-Mediated CRISPR Integration of Transgenes in Caenorhabditis elegans. G3: Genes|Genomes|Genetics, 9(8), 2629-2635.
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