Plan apochromat 1.4na

Manufactured by Zeiss

Sourced in Germany

The Plan-Apochromat 1.4NA from Zeiss is a high-numerical aperture (NA) objective lens designed for advanced microscopy applications. It features a numerical aperture of 1.4, which allows for the collection of a wide angle of light from the specimen, resulting in high-resolution imaging. The lens is also designed to provide a flat image field, ensuring consistent focus across the entire field of view.

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Plan-Apochromat (503 citations) Plan-Apochromat ×40/1.4NA oil objective (4 citations) Plan Apochromat 63x/1.4NA oil objective lens (3 citations) Plan-Apochromat 20X/1.4NA objective (2 citations) Plan apochromat 63X/1.4NA DIC M27 oil objective (2 citations) 100 x Plan Apochromat 1.4NA oil (2 citations) 63x Plan A apochromat 1.4NA oil objective (2 citations) 63× Plan-Apochromat 1.4NA DIC oil immersion objective (2 citations) 63x Plan-Apochromat 1.4na DIC objective (2 citations) Plan-Apochromat 100x/1.4NA oil-immersion objective (2 citations)

17 protocols using plan apochromat 1.4na

Immunostaining and Live-Cell Imaging Protocols

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For immunostaining, cells were cultured on coverslips and fixed in 4% formaldehyde. After washing twice with PBS, the cells were incubated in PBS (pH 7.4) containing 10% fetal calf serum (PBS/FCS) to block nonspecific sites of antibody adsorption. Then, the cells were incubated with appropriate primary and secondary antibodies in PBS/FCS buffer with 0.1% saponin as indicated in the figure legends. Images were captured with a Zeiss LSM510 Meta or LSM880 laser scanning confocal microscope (Carl Zeiss) with a 63× Plan Apochromat 1.4 NA objective.
For live-cell imaging, cells were cultured in chambers and imaged on a live-cell station. Photobleaching was performed using an appropriate laser line at full power. In FLIP analysis, a selected region was repetitively photobleached, and the loss of fluorescence from regions outside the photobleached region was monitored at low intensity illumination. For the FRAP analysis, a selected region of the cell was photobleached, and the fluorescence recovery of the region was monitored. The recovery half-time (t_1/2) was measured from the FRAP curve. All the FLIP and FRAP analyses were repeated at least five times in different cells. For quantification of fluorescence intensity, nonsaturated images were taken with a fully open pinhole, whereas nonquantitative images were obtained with a pinhole diameter equivalent to 1–2.5 Airy units.

Zhou Y., Wang Z., Huang Y., Bai C., Zhang X., Fang M., Ju Z, & Liu B. (2021). Membrane dynamics of ATG4B and LC3 in autophagosome formation. Journal of Molecular Cell Biology, 13(12), 853-863.

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Zeiss Confocal Microscopy Imaging Protocol

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Image acquisition was performed on a Zeiss LSM 710 or 780 confocal microscope (Carl Zeiss, Thornwood, NJ, USA) with a ×63 Plan-Apochromat 1.4 NA oil immersion objective using scan zoom of 2.5. Acquisition of Z-stacked images (voxel size of 0.105 μm × 0.105 μm × 0.34 μm) was at 512 × 512 pixels per frame using 8-bit pixel depth for each channel. The line averaging was set to 4, and images were collected sequentially in a three-channel mode.

Ranade D., Koul S., Thompson J., Prasad K.B, & Sengupta K. (2016). Chromosomal aneuploidies induced upon Lamin B2 depletion are mislocalized in the interphase nucleus. Chromosoma, 126(2), 223-244.

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Confocal Microscopy of Retina and Brain

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Confocal microscopy was performed on a Zeiss LSM 710 microscope. Overview images of the retina and brain were obtained with a 10x (plan-APOCHROMAT 0.45 NA, Zeiss) objective. The following settings were used: zoom 0.7, 4 × 4 tiles with 0% to 15% overlap, 2.37 µm/pixel resolution. For single retina ganglion cell scanning, we used a 63x (plan-APOCHROMAT 1.4 NA, Zeiss) objective. The following settings were used: zoom 0.7, 2 × 2 tiles or more (depending on size and number of cells) with 0% to 15% overlap. This resulted in an XY-resolution of 0.38 µm/pixel and a Z-resolution between 0.25 and 0.35 µm/pixel. The Z-stacks covered approximately 50 µm in depth.

Reinhard K., Li C., Do Q., Burke E.G., Heynderickx S, & Farrow K. (2019). A projection specific logic to sampling visual inputs in mouse superior colliculus. eLife, 8, e50697.

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Visualizing F-Actin Dynamics with Spinning Disk Confocal

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To visualize F-actin dynamics a Roper Spinning Disk Confocal System (Evry, France) consisting of a CSU-X1 spinning disk head (Yokogawa, Japan) mounted on a Nikon Eclipse Ti microscope (Tokyo, Japan) with Perfect Focus system was used. For imaging we used a 100× Plan-Apochromat 1.4 N.A. oil immersion objective. Fluorescence imaging was performed using a 491 nm laser line combined with band pass emission filtering (530/50 nm; Chroma Technology). Images were acquired with an Evolve electron-multiplying (EM) charge-coupled device camera (Photometrics) at an EM gain of 200, controlled by Metamorph software (Molecular Devices, California). Z-stacks were collected using a 0.5 μm step size.
For cell wall staining, solutions with calcofluor white (Sigma-Aldrich, Missouri) or aniline blue (Sigma-Aldrich, Missouri) were added to germinated cysts shortly before imaging to the final concentrations of 0.0017 and 0.2 % w/v, respectively. Fluorescence imaging was performed using a Zeiss LSM 510 Meta confocal microscope equipped with a 63× Plan-Apochromat 1.4 N.A. oil immersion objective. calcofluor white and aniline blue were imaged using the 405 nm laser line combined with a 420–480 nm band pass emission filter and a 420 nm long pass filter, respectively.

Kots K., Meijer H.J., Bouwmeester K., Govers F, & Ketelaar T. (2016). Filamentous actin accumulates during plant cell penetration and cell wall plug formation in Phytophthora infestans. Cellular and Molecular Life Sciences, 74(5), 909-920.

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Confocal Microscopy of Retina and Brain

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Confocal microscopy was performed on a Zeiss LSM 710 microscope. Overview images of the retina and brain were obtained with a 10× [plan-APOCHROMAT 0.45 NA (numerical aperture), Zeiss] objective. The following settings were used for imaging the whole-mount retina: zoom 0.7, 4 × 4 tiles with 0% to 15% overlap, 2.37 μm/pixel resolution. For single retinal ganglion cell scanning, we used a 63× (plan-APOCHROMAT 1.4 NA, Zeiss) objective. The following settings were used: zoom 0.7, 2 × 2 tiles or more (depending on size and number of cells) with 0% to 15% overlap. This resulted in an XY-resolution of 0.38 μm/pixel and a Z-resolution between 0.25 and 0.35 μm/pixel. The z-stacks covered approximately 50 μm in depth. The whole-brain images were acquired with the 10× objective with zoom 0.7, multiple tiles with 5% to 15% overlap, and a z-stack of 20 μm. To determine the axon bouton density in the visual TH and PBG, high-resolution z-stacks (0.24 × 0.24 μm²/pixel with 10 z-steps of 5 μm) of the cytosol and synaptophysin labeling were obtained with a 10× (plan-APOCHROMAT 0.45 NA, Zeiss) objective. The maximum projection of the z-stack was used for further analysis.

Li C., Kühn N.K., Alkislar I., Sans-Dublanc A., Zemmouri F., Paesmans S., Calzoni A., Ooms F., Reinhard K, & Farrow K. (2023). Pathway-specific inputs to the superior colliculus support flexible responses to visual threat. Science Advances, 9(35), eade3874.

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Transcriptional Dynamics via EU Labeling

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Transcription levels were determined following incubation with 2-hr ethynyluridine (EU)⁴⁰ added directly in the culture (serum-free) media. EU incorporation was visualized using Click-iT conjugation of Alexa Fluor 647 (Invitrogen) according to the manufacturer’s protocol. Images were obtained using a Zeiss Axio Imager Z2 upright laser-scanning confocal microscope equipped with a 63× Plan-Apochromat 1.4 NA oil immersion lens (Carl Zeiss Inc.) Fluorescence-signal intensities were quantified using the ImageJ software (NIH). In each experiment >150 cells per condition were analyzed.

Tresini M., Warmerdam D.O., Kolovos P., Snijder L., Vrouwe M.G., Demmers J.A., van IJcken W.F., Grosveld F.G., Medema R.H., Hoeijmakers J.H., Mullenders L.H., Vermeulen W, & Marteijn J.A. (2015). The core spliceosome as target and effector of non-canonical ATM signaling. Nature, 523(7558), 53-58.

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Confocal Imaging of Gut Samples

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Confocal images of guts were acquired on a Zeiss LSM 880 confocal microscope, using either a 20 × Plan-APOCHROMAT/0.8 numerical aperture (NA) or a 40 × oil Plan APOCHROMAT/1.4 NA objective. Confocal images are presented as maximum projections. Settings on the microscope were first adjusted on a control gut and maintained for the acquisition of the different conditions in each experiment. Images were analyzed in Fiji (Schindelin et al., 2012 (link)) or CellProfiler (Jones et al., 2008 ) and then linearly thresholded and assembled into figure panels in Photoshop Version 21.1.1 (Adobe Systems, San Jose, CA). All adjustments to contrast and other aspects of the images were performed similarly for all conditions in each experiment.

Nassari S., Lacarrière-Keïta C., Lévesque D., Boisvert F.M, & Jean S. (2022). Rab21 in enterocytes participates in intestinal epithelium maintenance. Molecular Biology of the Cell, 33(4), ar32.

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Confocal Imaging of Protein Interactions

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N. benthamiana leaf samples (approximately 0.25 cm² near the infiltrated area) were collected at 70–90 h post-infiltration, mounted in water and viewed directly with a Zeiss LSM 880 confocal scanning microscope using an oil immersion objective 40× Plan-Apochromat 1.4NA (numerical aperture of 1.4). Fluorescence was excited using 488 nm and 543 nm light for GFP and RFP, respectively. GFP and RFP emission fluorescence was selectively detected at 490–540 and 550–630 nm using the Zen 2.3 SP1 software. For each experiment, 50–100 cells in two independent leaves that expressed both proteins were evaluated. For the BiFC experiments, YFP fluorescence was excited using 514 nm laser and detected at 520–550 nm using the Leica TCS SP5 confocal scanning microscope with a 25x objective.

Wu S., Zhang B., Keyhaninejad N., Rodríguez G.R., Kim H.J., Chakrabarti M., Illa-Berenguer E., Taitano N.K., Gonzalo M.J., Díaz A., Pan Y., Leisner C.P., Halterman D., Buell C.R., Weng Y., Jansky S.H., van Eck H., Willemsen J., Monforte A.J., Meulia T, & van der Knaap E. (2018). A common genetic mechanism underlies morphological diversity in fruits and other plant organs. Nature Communications, 9, 4734.

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Transcriptional Dynamics via EU Labeling

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Imaging GFP in N. benthamiana Leaves

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N. benthamiana leaf samples 70 to 90 hours post-infiltration (approximately 0.25 cm² from the infiltrated area) were mounted in water and viewed directly with a Zeiss LSM 880 confocal scanning microscope using an oil immersion objective 40× Plan-Apochromat 1.4NA (numerical aperture of 1.4). Fluorescence was excited using 488 nm light for GFP. GFP emission fluorescence was selectively detected at 490–540 nm using the Zen 2.3 SP1 software. Each experiment was repeated three times.

Shrestha S., Katiyar S., Sanz-Rodriguez C.E., Kemppinen N.R., Kim H.W., Kadirvelraj R., Panagos C., Keyhaninejad N., Colonna M., Chopra P., Byrne D.P., Boons G.J., van der Knaap E., Eyers P.A., Edison A.S., Wood Z.A, & Kannan N. (2020). A redox-active switch in fructosamine-3-kinases expands the regulatory repertoire of the protein kinase superfamily. Science signaling, 13(639), eaax6313.

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