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Plan apochromat 100 1.40 n a

Manufactured by Zeiss
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The Plan-Apochromat 100×/1.40 N.A. is a high-performance objective lens designed for microscopy applications. It features a numerical aperture of 1.40 and a magnification of 100x, providing a high resolution and light-gathering capability. The lens is part of the Plan-Apochromat series, known for its excellent optical performance and flatness of field.

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Lab products found in correlation

Lsm 710, by Zeiss (4 mentions) Cell light fluorescent proteins, by Thermo Fisher Scientific (2 mentions) Axioskop 2 mot, by Zeiss (1 mentions) Axiocam mrm, by Zeiss (1 mentions) Axiovision software, by Zeiss (1 mentions) Orca er camera, by Hamamatsu Photonics (1 mentions) Matlab, by MathWorks (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Eclipse ti u microscope, by Nikon (1 mentions) Lsm510 axiovert 200m microscope, by Zeiss (1 mentions)

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Plan-Apochromat (503 citations) Plan Apochromat 100 (36 citations)

5 protocols using plan apochromat 100 1.40 n a

1

Quantifying APP Enrichment in Golgi Apparatus

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Fluorescent images were taken using a confocal microscope (Zeiss LSM 710) with an oil immersion lens (Plan-Apochromat 100 ×/1.40 N.A. Oil DIC) and a 2 × digital zoom. DAPI was excited using the 405 nm emitting diode laser; Alexa Fluor 488 or organelle markers labeled with GFP (Cell Light fluorescent proteins, Invitrogen) using an argon laser (488 nm); Alexa Fluor 568 dye using a 561 nm DPSS laser; Alexa Fluor 633 and 647 dyes using a HeNe laser (633 nm). To obtain line scans to test for co-localization, the fluorescence intensity signals for WAVE1, APP and Golgi-GFP were plotted against distance in μm for all three colors (Fig. 3c, indicated by the white line). An average of 5 pixels was used to obtain line scans for the data shown in Supplemental Fig. 4 and plotted against distance in μm. For the quantification of APP in the Golgi apparatus, the z slice with the strongest Golgi signal was chosen. The images were analyzed with the MetaMorph software (ver. 7.7.8). The immunostaining of a Golgi marker protein, giantin, was used to define a mask for the Golgi area to quantify the mean intensity of APP signal in this area. One cytoplasmic area of the same dimension as the Golgi area was randomly chosen. The ratio of the average intensity of APP signal on Golgi over the average intensity of APP signal in the cytoplasm was determined as APP enrichment.
Ceglia I., Reitz C., Gresack J., Ahn J.H., Bustos V., Bleck M., Zhang X., Martin G., Simon S.M., Nairn A.C., Greengard P, & Kim Y. (2015). APP intracellular domain/WAVE1 pathway reduces amyloid β production. Nature medicine, 21(9), 1054-1059.
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2

Fluorescence Microscopy for Peroxisome Localization

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0.1 ml of cell culture was collected and kept on ice before observation. For slides, 5 µl of cell culture was mixed with 5 µl of 1% low-melt agarose kept at 37°C. Images were captured at RT using a motorized fluorescence microscope (Axioskop 2 MOT) with a Plan-Apochromat 100×/1.40 NA oil differential interference contrast (DIC) objective lens and monochrome digital camera (AxioCam MRm; all from Carl Zeiss). Optimal exposition times were automatically applied to capture images. Z-stack images (25 slices, 0.255 µm apart) were used to prove Atg37-GFP localization in the middle of the peroxisome cluster. All images were acquired and processed using the AxioVision software (Carl Zeiss), versions 4.7.1 and 4.8.2, respectively. Additionally, we used version 10.0 of Photoshop CS3 Extended (Adobe).
Nazarko T.Y., Ozeki K., Till A., Ramakrishnan G., Lotfi P., Yan M, & Subramani S. (2014). Peroxisomal Atg37 binds Atg30 or palmitoyl-CoA to regulate phagophore formation during pexophagy. The Journal of Cell Biology, 204(4), 541-557.
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3

Microscopy Techniques for Cell Analysis

Cited 1 time
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For all microscopy observations, cells were spotted on 1% agarose pads containing M2G medium, unless specified otherwise. Images were acquired using an Eclipse Ti-U microscope (Nikon, Tokyo, Japan) with an Orca-ER camera (Hamamatsu Photonics, Hamamatsu City, Japan) and phase-contrast objective Plan Apochromat 100 × /1.40 NA (Carl Zeiss, Oberkochen, Germany) at room temperature except for time-lapse experiments (30°C). Images were acquired and processed with either MetaMorph software (Molecular Devices, Sunnyvale, CA) or MATLAB (The MathWorks, Natick, MA). Cell mesh creation and fluorescence quantification were done using the open source, MATLAB-based software MicrobeTracker (Sliusarenko et al., 2011 (link)). Identification of diffraction-limited fluorescence spots was done using SpotFinder, an accessory in MicrobeTracker, unless mentioned otherwise.
Lim H.C., Surovtsev I.V., Beltran B.G., Huang F., Bewersdorf J, & Jacobs-Wagner C. (2014). Evidence for a DNA-relay mechanism in ParABS-mediated chromosome segregation. eLife, 3, e02758.
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4

Quantifying APP Enrichment in Golgi Apparatus

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Fluorescent images were taken using a confocal microscope (Zeiss LSM 710) with an oil immersion lens (Plan-Apochromat 100 ×/1.40 N.A. Oil DIC) and a 2 × digital zoom. DAPI was excited using the 405 nm emitting diode laser; Alexa Fluor 488 or organelle markers labeled with GFP (Cell Light fluorescent proteins, Invitrogen) using an argon laser (488 nm); Alexa Fluor 568 dye using a 561 nm DPSS laser; Alexa Fluor 633 and 647 dyes using a HeNe laser (633 nm). To obtain line scans to test for co-localization, the fluorescence intensity signals for WAVE1, APP and Golgi-GFP were plotted against distance in μm for all three colors (Fig. 3c, indicated by the white line). An average of 5 pixels was used to obtain line scans for the data shown in Supplemental Fig. 4 and plotted against distance in μm. For the quantification of APP in the Golgi apparatus, the z slice with the strongest Golgi signal was chosen. The images were analyzed with the MetaMorph software (ver. 7.7.8). The immunostaining of a Golgi marker protein, giantin, was used to define a mask for the Golgi area to quantify the mean intensity of APP signal in this area. One cytoplasmic area of the same dimension as the Golgi area was randomly chosen. The ratio of the average intensity of APP signal on Golgi over the average intensity of APP signal in the cytoplasm was determined as APP enrichment.
Ceglia I., Reitz C., Gresack J., Ahn J.H., Bustos V., Bleck M., Zhang X., Martin G., Simon S.M., Nairn A.C., Greengard P, & Kim Y. (2015). APP intracellular domain/WAVE1 pathway reduces amyloid β production. Nature medicine, 21(9), 1054-1059.
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5

Subcellular Localization of ORP1L, LAMP1/2, and LDLR

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Cells were lysed in SDS sample buffer (12.5% SDS, 30 mM Tris-HCl, 12.5% glycerol, and 0.01% bromophenol blue, pH 6.8), heated at 90°C for 5 min, separated by SDS–PAGE, and transferred to nitrocellulose. The following primary antibodies were used: ORP1L (1:2000, Abcam, ab131165), LAMP1 and LAMP2 (1:1000, Developmental Studies Hybridoma Bank, 1D4B and H4B4), LDLR (1:2000, rabbit polyclonal Ab3143 targeting the C-terminus) (Russell et al., 1984 ), or actin (1:10,000, Sigma AC15). Proteins were visualized with IRDye-800– or -680–conjugated secondary antibodies using an Odyssey Imaging System and application software v3.0 (LI-COR Biosciences).
Cells cultured on glass coverslips were fixed in 4% (wt/vol) paraformaldehyde and permeabilized with 0.05% (wt/vol) Triton X-100 at 4°C. Cells were probed with LAMP1 or LDLR antibodies and appropriate secondary Alexa Fluor–conjugated antibodies (1:5000, ThermoFisher) in phosphate-buffered saline (PBS) containing 1% (wt/vol) BSA. Confocal imaging was performed using a Zeiss LSM510/Axiovert 200M microscope with a Plan-Apochromat 100×/1.40 NA oil immersion objective.
Zhao K., Foster J, & Ridgway N.D. (2020). Oxysterol-binding protein-related protein 1 variants have opposing cholesterol transport activities from the endolysosomes. Molecular Biology of the Cell, 31(8), 793-802.
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