Plan apochromat 63 1.4 na oil

Manufactured by Zeiss

Sourced in Germany

The Plan-Apochromat 63× 1.4 NA oil is a high-performance objective lens designed for use in microscopy applications. It features a magnification of 63× and a numerical aperture of 1.4, which enables high-resolution imaging and excellent light-gathering capabilities. The lens is optimized for use with oil immersion to provide superior optical performance.

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Plan Apochromat 63×1.4 NA (oil/dic) objective Plan Apochromat 63×/1.4 NA oil immersion lens Plan-apochromat 63 × 1.4 NA oil objective lens Plan-Apochromat ×63/1.4 NA M27 oil immersion objective Plan-Apochromat 63×/1.4 NA DIC M27 Oil objective Plan-Apochromat 63×/1.4-NA oil lens Plan Apochromat 63×/1.4 NA oil immersion objective Plan-Apochromat 63×/1.4 NA oil objective Plan-apochromat X 63 (NA 1.4) oil immersion objective Plan-Apochromat 63×/1.4 NA

14 protocols using plan apochromat 63 1.4 na oil

Mitochondrial Staining and Imaging

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MEFs and 786-O cells were cultured on glass coverslips and stained with MitoTracker Red CMXRos (100 nM) at 37 °C for 30 min, washed twice with pre-warmed PBS, and fixed for 15 min in pre-warmed 4% paraformaldehyde. Coverslips were immersed into PBS overnight, and mounted using ProLong Diamond Antifade Mountant with DAPI (Thermo Fisher, P36962). Fluorescence images were acquired using a Zeiss LSM 700 laser scanning confocal microscope equipped with a ×63 Plan-Apochromat/1.4-NA Oil with DIC capability objective. The excitation wavelengths for MitoTracker Red CMXRos and DAPI were 579 nm and 405 nm, respectively. Images were acquired using the settings: frame size of 1,024, scan speed of 6 and 12-bit acquisition and line averaging mode of 8. Pinholes were adjusted so that each channel had the same optical slice of 1–1.2 μm.

Li S., Li W., Yuan J., Bullova P., Wu J., Zhang X., Liu Y., Plescher M., Rodriguez J., Bedoya-Reina O.C., Jannig P.R., Valente-Silva P., Yu M., Henriksson M.A., Zubarev R.A., Smed-Sörensen A., Suzuki C.K., Ruas J.L., Holmberg J., Larsson C., Christofer Juhlin C., von Kriegsheim A., Cao Y, & Schlisio S. (2022). Impaired oxygen-sensitive regulation of mitochondrial biogenesis within the von Hippel–Lindau syndrome. Nature Metabolism, 4(6), 739-758.

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Mitochondrial Staining and Imaging Procedure

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786-O and MEFs cells were cultured on glass coverslips and stained with
MitoTracker Red CMXRos (100 nM) at 37 °C for 30 min, washed twice with
pre-warmed phosphate-buffered saline (PBS), and fixed for 15 min in pre-warmed
4% paraformaldehyde. Coverslips were immersed into PBS overnight, and mounted
using ProLong Diamond Antifade Mountant with DAPI (ThermoFisher, cat no:
P36962). Fluorescence images were acquired using a Zeiss LSM 700 laser scanning
confocal microscope equipped with a 63× Plan-Apochromat/1.4 NA Oil with
DIC capability objective. The excitation wavelengths for MitoTracker Red CMXRos
and DAPI were 579 nm and 405 nm, respectively. Images were acquired under the
settings: frame size 1024, scan speed 6, and 12-bit acquisition and line
averaging mode 8. Pinholes were adjusted so that each channel had the same
optical slice of 1-1.2 μm.

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Multimodal Confocal Microscopy Imaging

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Images were acquired with a Zeiss LSM 510 Duoscan laser-scanning confocal microscope or a Zeiss LSM 780 NLO 2-photon laser-scanning microscope with an automated tuning pulsed Ti:S Chameleon Vision II laser (Coherent), using 63× Plan-Apochromat 1.4 N.A. oil or 40× C-Apochromat 1.2 N.A. W objectives (Carl Zeiss, Germany). Multiposition and multidimensional imaging was controlled as described (Rabut and Ellenberg, 2004 (link)).

Mora-Bermúdez F., Matsuzaki F, & Huttner W.B. (2014). Specific polar subpopulations of astral microtubules control spindle orientation and symmetric neural stem cell division. eLife, 3, e02875.

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Confocal Imaging of HCV Infection

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Images were acquired on a Zeiss LSM 510 confocal microscope (Zeiss NLO 510 with META; Zeiss Plan-Apochromat 63/1.4NA oil; Thornwood, NY). Imaging settings were defined empirically to maximize the signal-to-noise ratio and to avoid saturation. In comparative imaging, all settings were kept constant between samples. The illumination was provided by 30 mW Argon (488 nm), 5 mW HeNe (633 nm) and 1 mW HeNe (543 nm) lasers. Image processing was performed using Zeiss ZEN 2009 software. Figures were mounted using Adobe Photoshop CS4 (Adobe System).
For the experimental setup, cells were seeded onto 35 mm glass bottom dishes (MatTek Corporation, Ashland, MA) and infected on day 1 and 7 with JFH1 at multiplicity of infection of 10 (MOI of 10). After 14 days of infection cells were fixed in 4% PFA in PBS. Cells were then blocked with 10% normal serum, labeled with primary polyclonal antibodies against HCV-NS5A (clone 9E10, 1:100, provided by C. Rice, Rockefeller University) and EEA1 (rabbit polyclonal 1:200, Abcam), and washed and labeled with the appropriate secondary antibody that was conjugated to Alexa-Fluor 633 or Alexa-Fluor 568 (Invitrogen). F-actin was concurrently stained with Alexa-Phalloidin 546 (Invitrogen). Images were acquired as described above.

Giugliano S., Petroff M.G., Warren B.D., Jasti S., Linscheid C., Ward A., Kramer A., Dobrinskikh E., Sheiko M.A., Gale M J.r., Golden-Mason L., Winn V.D, & Rosen H.R. (2015). HCV Sensing by Human Trophoblasts Induces Innate Immune Responses and Recruitment of Maternal NK Cells: Potential Implications for Limiting Vertical Transmission. Journal of immunology (Baltimore, Md. : 1950), 195(8), 3737-3747.

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High-Resolution Fluorescence Microscopy

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Images were acquired as 0.3 µm z stacks using a DeltaVision microscope (Applied Precision Ltd.) or a confocal laser-scanning microscope (LSM780; ZEISS) with a Plan-Apochromat 63× 1.4 NA oil differential interference contrast objective equipped with two photomultiplier tubes and a gallium arsenite phosphate (GaAsP-PMT) detector system. SoftWoRx software (GE Healthcare) was used for acquisition and deconvolution (Applied Precision Ltd.) of images acquired with the DeltaVision microscope, and ZEN 2012 (black edition; 8.0.5.273; ZEISS) was used for acquisition of images with the LSM780 microscope. Deconvolved images were processed and analyzed with FIJI (ImageJ; National Institutes of Health).

Botti V., McNicoll F., Steiner M.C., Richter F.M., Solovyeva A., Wegener M., Schwich O.D., Poser I., Zarnack K., Wittig I., Neugebauer K.M, & Müller-McNicoll M. (2017). Cellular differentiation state modulates the mRNA export activity of SR proteins. The Journal of Cell Biology, 216(7), 1993-2009.

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Optimizing Confocal Imaging Protocols

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Confocal microscopy settings were always kept constant throughout the same set of experiments. Zeiss LSM5 Pascal confocal microscope was used for all axonal debris clearance, TRE-eGFP reporter (except for UAS-msn^DN), and STAT92E reporter assays with Zeiss Plan-Apochromat × 63/1.4NA oil objective lens. Glial MARCM clones, glial activation in hiw^ΔN animals and TRE-eGFP reporter activity in msn^DN animals were analysed using Zeiss Axio Imager.M2 spinning-disk confocal microscope with Zeiss C-Apochromat × 40/1.2NA water objective lens. For quantification of the fluorescence intensity, the centre single z-section was identified from each image stack of the relevance glomerulus or central adult brain, and pixel intensity was measured using ImageJ (National Institute of Health).

Lu T.Y., MacDonald J.M., Neukomm L.J., Sheehan A.E., Bradshaw R., Logan M.A, & Freeman M.R. (2017). Axon degeneration induces glial responses through Draper-TRAF4-JNK signalling. Nature Communications, 8, 14355.

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Imaging Drosophila Larval Dendritic Arbors

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3-day-old drosophila larvae were mounted with a drop of water onto an agarose pad on the surface of a glass slide that allows immobilization of the larva. A coverslip was placed on the larva and taped to the slide such that the larva is lightly pressed and has the dorsal side facing upwards. The ddaC neurons in the a2 or a3 segment were chosen for imaging for consistency. In all the experiments, only one neuron was imaged from one larva/animal.
All Z-stack images/ time-lapse videos were acquired using a Zeiss LSM800 upright confocal microscope with a Plan-Apochromat 63× 1.4 NA oil objective unless otherwise mentioned. EB1-TagRFP-T movies were 150 frames at one frame per two seconds, so total duration of five minutes. These movies were taken at 0.5x zoom in order to cover most of the dendritic arbor. Time-lapse videos of EB1-GFP were taken with a widefield Zeiss Imager M2 microscope with AxioCam M2 camera and 63× 1.4 NA Plan-Apochromat oil immersion objective. Zeiss Zen Blue software was used to record 300-frames at the speed of one frame per second, so duration of five minutes. Figure 4F was generated using the AiryScan detector and processing on a Zeiss LSM800 upright microscope.

Thyagarajan P., Feng C., Lee D., Shorey M, & Rolls M.M. (2022). Microtubule polarity is instructive for many aspects of neuronal polarity. Developmental biology, 486, 56-70.

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Multicolor Fluorescence Microscopy Protocol

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Microscopy was performed on a Carl Zeiss Axiovert 200 Μ with a Plan-Apochromat 63×/1.4NA oil lens and wavelength filter sets for detection of Hoechst 33342 (450 nm), Alexa Fluor 488 (520 nm) and Alex Fluor 555 (568 nm) The images were acquired using the high sensitivity digital camera AxioCam HRc for multicolor fluorescent images along with the Zeiss software Axiovision, v4.8.2.SP2. All same-figure images were taken at identical exposure and filter settings for comparability of intensities. Images were processed with uniform and unbiased application of contrast and brightness settings in Photoshop and Illustrator (Adobe Inc., San Jose, CA, USA).

Maimaris G., Christodoulou A., Santama N, & Lederer C.W. (2021). Regulation of ER Composition and Extent, and Putative Action in Protein Networks by ER/NE Protein TMEM147. International Journal of Molecular Sciences, 22(19), 10231.

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Confocal Microscopy Imaging Protocol

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Confocal micrographs were acquired on a Zeiss LSM 710 Laser Scanning Microscope equipped with a Plan Apochromat 63×/ 1.4 NA oil immersion objective. ZEN software was used to collect the data. Images were analyzed with Image J.

Li C., Plamont M.A., Aujard I., Le Saux T., Jullien L, & Gautier A. (2016). Design and characterization of red fluorogenic push-pull chromophores holding great potential for bioimaging and biosensing. Organic & biomolecular chemistry, 14(39).

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Red Blood Cell Membrane Protein Labeling

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Diluted washed RBCs were spread onto PLL-coated coverslips for 4 min, fixed with 4% (v/v) paraformaldehyde for 10 min and blocked with 1% (w/v) BSA for 30 min. To label membrane proteins, RBCs were then incubated for 1 h with rabbit monoclonal antibodies to glycophorin C (GPC) together with mouse monoclonal antibodies to CD47 (Invitrogen), washed 4 times in 1% (w/v) BSA, incubated for 1 h with the appropriate Alexa-secondary antibodies (5 µg/mL) and washed 4 times with 1% BSA. Total spectrin was revealed with antibodies against α/β-spectrins (Abcam) using the same protocol as above except that a permeabilization step with 0.5% (w/v) Triton X-100 for 3 min was done before the fixation. All coverslips were mounted in Mowiol in the dark for 24 h and examined with a Zeiss LSM510 confocal microscope using a plan-Apochromat 63× NA 1.4 oil immersion objective.

Pollet H., Cloos A.S., Stommen A., Vanderroost J., Conrard L., Paquot A., Ghodsi M., Carquin M., Léonard C., Guthmann M., Lingurski M., Vermylen C., Killian T., Gatto L., Rider M., Pyr dit Ruys S., Vertommen D., Vikkula M., Brouillard P., Van Der Smissen P., Muccioli G.G, & Tyteca D. (2020). Aberrant Membrane Composition and Biophysical Properties Impair Erythrocyte Morphology and Functionality in Elliptocytosis. Biomolecules, 10(8), 1120.

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