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Na 1.4 oil objective

Manufactured by Zeiss
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The 63x/NA 1.4 oil immersion objective from Zeiss is a high-magnification lens designed for microscopy applications. It offers a numerical aperture of 1.4, providing a high resolution and light-gathering capability. The objective is optimized for use with oil immersion, which can further enhance image quality.

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Lab products found in correlation

Zen blue software, by Zeiss (4 mentions) X tremegene 9, by Merck Group (2 mentions) Plan apochromat, by Zeiss (2 mentions) Lambda ls xenon arc lamp light source system, by Sutter Instruments (2 mentions) Observer z1 microscope, by Zeiss (2 mentions) Axiocam mrm camera, by Zeiss (2 mentions) Bbig l1, by R&D Systems (1 mentions) Lsm 510 meta confocal laser scanning microscope, by Zeiss (1 mentions) Polystyrene beads, by Polysciences (1 mentions) Imagej, by GraphPad (1 mentions) Lsm 510 meta confocal laser, by Zeiss (1 mentions) Prism 5, by GraphPad (1 mentions) Zyla 4.2 scmos camera, by Oxford Instruments (1 mentions) Mosaic digital micromirror device, by Oxford Instruments (1 mentions) Metamorph automation and image analysis software, by Molecular Devices (1 mentions) Airyscan 2, by Zeiss (1 mentions) Csu x confocal scanning head, by Yokogawa (1 mentions) Pma hybrid 40 detector, by PicoQuant (1 mentions)

Close spelling variants of this product

Plan-Apochromat 63×/1.4 NA oil objective Plan- Apochromat 40× NA-1.4 oil objective 63× 1.4 NA oil objective lens Plan-Apochromat 100x/1.4 (NA) Oil DIC objective Plan-Apochromat 63×/1.4 NA DIC M27 Oil objective Plan-Apochromat 1.4 NA 100x oil objective Plan-apochromat X 63 (NA 1.4) oil immersion objective Plan Apochromat 63×1.4 NA (oil/dic) objective Zeiss x63 1.4-NA Oil Plan-Apochromat objective 63× PLAN APO (1.4 NA) oil-immersion objectives

6 protocols using na 1.4 oil objective

1

Protein Recruitment to Anti-ICAM-1 Antibody Beads

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To analyze protein recruitment to anti-ICAM-1-antibody-coated beads, 10 μm polystyrene beads (Polysciences) were coated with mouse monoclonal antibody anti-ICAM-1 (BBIG-l1, R&D Systems) using glutaraldehyde according to the manufacturer’s protocol (16 (link)). Beads were incubated with a TNFα-stimulated HUVEC monolayer for 15 min at 37°C and 5% CO2. Cells were washed twice with PBS containing 1 mM CaCl2 and 0.5 mM MgCl2 to remove unbound beads, fixed and immunostained. Images were recorded in six random fields with a Zeiss LSM510-META confocal laser scanning microscope (63x/NA 1.4 oil objective) with z-sections at two positions (1.5 μm interval). Protein recruitment was counted as positive if at least half of the adherent bead at the apical plane was surrounded by the fluorescence signal of the immunostained protein. Percentage of beads with ring formation was calculated as (number of beads with ring formation)/total number of adherent beads*100. Numbers were then normalized to the si-Control condition which was set to 1.
Schaefer A., van Duijn T.J., Majolee J., Burridge K, & Hordijk P.L. (2017). Endothelial CD2AP binds the receptor ICAM-1 to control mechanosignaling, leukocyte adhesion and the route of leukocyte diapedesis in vitro. Journal of immunology (Baltimore, Md. : 1950), 198(12), 4823-4836.
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2

Live imaging of ICAM-1 recruitment

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To study recruitment to anti-ICAM-1-antibody-coated beads, TNFα-treated HUVECs were transfected as indicated and cultured on fibronectin-coated 30 mm glass coverslips (Thermo Fisher Scientific) and placed in a heating chamber at 37°C and 5% CO2. GFP- and DIC-signals were monitored using a Zeiss LSM-510-META confocal laser scanning microscope (63x/NA 1.4 oil objective). 10 μm polystyrene beads coated with anti-ICAM-1 antibody (see above) were added after 2 min and imaging was performed for up to 23 min with intervals of 24s (ICAM1-GFP vs CD2AP-GFP) or 20s (ICAM-1-GFP in knock-down studies) or 10.3s (Lifeact-GFP) and with z-sections at three positions (2.5 μm intervals). Fluorescence intensity was quantified in a donut-shaped area positioned at the center of the bead and corrected for background, bleaching and transfection efficiency for each cell, time point and confocal plane using Zen2009 and Zen2 software, ImageJ and Prism5 (Graphpad) (16 (link)) For normalization, the average of the saturated signal was calculated as 100% and the time point of adding the beads as 0 s.
Schaefer A., van Duijn T.J., Majolee J., Burridge K, & Hordijk P.L. (2017). Endothelial CD2AP binds the receptor ICAM-1 to control mechanosignaling, leukocyte adhesion and the route of leukocyte diapedesis in vitro. Journal of immunology (Baltimore, Md. : 1950), 198(12), 4823-4836.
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3

Optoregulation of Cellular Dynamics

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Optogenetic experiments were performed on an inverted Nikon T-E (Nikon, Melville, NY) with a laser merge module with 491, 561, and 642nm laser lines (Spectral Applied Research, Ontario, Canada) with a Yokogawa CSU-X confocal scanning head (Yokogawa Electric, Tokyo, Japan). The Zyla 4.2 sCMOS Camera (Andor, Belfast, UK) collected the images. Optogenetic activation was achieved using a Mosaic digital micromirror device (Andor) attached to a 405nm laser. Images were collected on a 60X 1.2 Plan Apo water (Nikon) objective. MetaMorph Automation and Image Analysis Software (Molecular Devices, Sunnyvale, CA) controlled all hardware. Fix-and-stain and live-cell imaging of CN03 wash-ins were performed on an LSM 980 system with an Airyscan 2 (Zeiss) detector in super resolution-mode with a 63x NA1.4 oil objective (Zeiss). Microscopy software used was the Zen digital imaging suite (Zeiss).
Cavanaugh K.E., Staddon M.F., Chmiel T.A., Harmon R., Budnar S., Yap A.S., Banerjee S, & Gardel M.L. (2022). Force-dependent intercellular adhesion strengthening underlies asymmetric adherens junction contraction. Current biology : CB, 32(9), 1986-2000.e5.
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4

Measuring Membrane Protein Dynamics

Cited 1 time
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3T3 cells stably expressing GBP-TM-VSVG-mScarlet were spread on fibronectin-coated glass-bottom dishes, and the transmembrane construct was then clustered with B(c)-GFP and A(d) as described above, before mitotic stalling was induced with 30 nM nocodazole for 12 h. The Flipper-TR probe (Spirochrome, see Colom et al.22 (link)) was then added (2 μM final in L15-20 mM HEPES medium) and Flipper-TR fluorescence lifetime imaging was performed on a setup comprising a Zeiss LSM710 stand, a 63X NA 1.4 oil objective, a Zeiss 710 confocal scanner head and Time-Correlated Single Photon Counting (TCSPC) hardware from Picoquant. A 470 nm pulsed laser (Picoquant), operating at 40 MHz was used to excite the probe, and detection was performed on a gated PMA hybrid 40 detector (Picoquant) behind a 600/50 bandpass filter (Semrock). SymPhotime 2.0 software (Picoquant) was used for data analysis. Flipper-TR fluorescence lifetime was fitted to a dual exponential model. The intensity-weighted average lifetime in a Regions of Interest (ROI) encompassing either the array-containing membrane (assessed by GFP signal), or regions of the membrane devoid of arrays was then measured, followed by averaging over several cells.
Watson J.L., Krüger L.K., Ben-Sasson A.J., Bittleston A., Shahbazi M.N., Planelles-Herrero V.J., Chambers J.E., Manton J.D., Baker D, & Derivery E. (2023). Synthetic Par polarity induces cytoskeleton asymmetry in unpolarized mammalian cells. Cell, 186(21), 4710-4727.e35.
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5

FRET Imaging of Transfected HEK293-FT Cells

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Cultured HEK293-FT cells were trypsinized ≤ 2 min, counted, and 75,000 cells were seeded onto uncoated 18 mm glass coverslips. After 24 h growth, cells were transfected using OPTI-MEM serum free medium, X-tremeGene 9 (Sigma, 6365779001) and various plasmids. Living cells were imaged in a Tyrode’s salt solution (in mM: 135 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 25 HEPES, 10 glucose, pH 7.4) at RT 24–48 h post-transfection. An Observer.Z1 microscope (Zeiss) with a 63x plan-apochromat, 1.4 NA oil objective (Zeiss), Lambda LS Xenon Arc Lamp Light Source System (Sutter Instruments), AxioCam MRm camera (Zeiss), and Zen Blue software (Zeiss) were used for image acquisition. Three-filter FRET images were captured using appropriate filter cubes (Semrock) housed in the microscope turret. CFP cube: (Ex. 438/24 nm, Em. 483/32 nm, Di. 458 nm), YFP cube: (Ex. 500/24 nm, Em. 542/27 nm, Di. 520 nm), and FRET cube: (Ex. 438/24 nm, Em. 542/27 nm, Di. 458 nm). ImageJ software (NIH) was used for image processing and calculations of sensitized FRET efficiency were adapted from the method of Clemens Kaminski (Kaminski et al., 2014 ) with more details provided below.
Murphy J.G., Gutzmann J.J., Lin L., Hu J., Petralia R.S., Wang Y.X, & Hoffman D.A. (2022). R-type voltage-gated Ca2+ channels mediate A-type K+ current regulation of synaptic input in hippocampal dendrites. Cell reports, 38(3), 110264.
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6

FRET Imaging of Transfected HEK293-FT Cells

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Cultured HEK293-FT cells were trypsinized ≤ 2 min, counted, and 75,000 cells were seeded onto uncoated 18 mm glass coverslips. After 24 h growth, cells were transfected using OPTI-MEM serum free medium, X-tremeGene 9 (Sigma, 6365779001) and various plasmids. Living cells were imaged in a Tyrode’s salt solution (in mM: 135 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 25 HEPES, 10 glucose, pH 7.4) at RT 24–48 h post-transfection. An Observer.Z1 microscope (Zeiss) with a 63x plan-apochromat, 1.4 NA oil objective (Zeiss), Lambda LS Xenon Arc Lamp Light Source System (Sutter Instruments), AxioCam MRm camera (Zeiss), and Zen Blue software (Zeiss) were used for image acquisition. Three-filter FRET images were captured using appropriate filter cubes (Semrock) housed in the microscope turret. CFP cube: (Ex. 438/24 nm, Em. 483/32 nm, Di. 458 nm), YFP cube: (Ex. 500/24 nm, Em. 542/27 nm, Di. 520 nm), and FRET cube: (Ex. 438/24 nm, Em. 542/27 nm, Di. 458 nm). ImageJ software (NIH) was used for image processing and calculations of sensitized FRET efficiency were adapted from the method of Clemens Kaminski (Kaminski et al., 2014 ) with more details provided below.
Wood G.E., Dutro S.M, & Totten P.A. (1999). Target Cell Range of Haemophilus ducreyi Hemolysin and Its Involvement in Invasion of Human Epithelial Cells. Infection and Immunity, 67(8), 3740-3749.
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