HEK293T cells were cultured in Dulbecco's modified Eagle’s medium (DMEM, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS) at 37°C and 5% CO2. For receptor localization analysis, HEK293T cells were seeded in 35-mm sterilized glass-bottom dishes and incubated for 24 h. EGFP-GR66-pcDNA3.0 was transfected into HEK293T cells using Lipofectamine 2000 (Invitrogen). After 24 h, the cells were fixed with 4% paraformaldehyde for 15 min and finally incubated with DAPI for 10 minutes. The cells were visualized by fluorescence microscopy on a Zeiss LSM 510 confocal laser scanning microscope attached to a Zeiss Axiovert 200 microscope using a Zeiss Plan-Apochromat 63×/1.40 NA oil immersion lens.
Plan apochromat 63
Manufactured by Zeiss
Sourced in Germany
The Plan-Apochromat 63 is a high-performance objective lens designed for microscopy applications. It features a plan-apochromatic optical design, ensuring a flat field of view and accurate color reproduction across the entire field. The lens has a numerical aperture of 1.40, providing high resolution and light-gathering capabilities. It is suitable for a variety of microscopy techniques, including brightfield, fluorescence, and differential interference contrast (DIC) imaging.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1, by Zeiss (9 mentions)
Lsm 710, by Zeiss (8 mentions)
Lsm 510, by Zeiss (7 mentions)
Lsm 510 meta, by Zeiss (6 mentions)
Lsm 780 confocal microscope, by Zeiss (6 mentions)
Zen software, by Zeiss (4 mentions)
Lsm800 confocal microscope, by Zeiss (4 mentions)
Lsm 780, by Zeiss (4 mentions)
Lsm 880, by Zeiss (4 mentions)
Lsm 900 confocal microscope, by Zeiss (3 mentions)
Imagej, by Adobe (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Axiocam digital camera, by Zeiss (2 mentions)
Lsm image browser, by Zeiss (2 mentions)
Zen 2, by Zeiss (2 mentions)
Hoechst, by Thermo Fisher Scientific (2 mentions)
Axiovert 200m, by Zeiss (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Axiovert 200 microscope, by Zeiss (1 mentions)
44 protocols using plan apochromat 63
1
Localization of GR66 in HEK293T Cells
2
Halo-Myh3 Protein Synthesis Tracking
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Day 6 myotubes expressing Halo‐Myh3 were labeled with Oregon‐Green®︎ ligand (green fluorescence; Promega) at a final concentration of 0.3% in DM for 16 h. Following washout, myotubes were reacted with TMR ligand (red fluorescence, Promega) at a final concentration of 0.1% in minimum essential medium (Thermo Fisher Scientific) to label newly synthesized Halo‐Myh3 for 7, 15, and 30 min. Myotubes were fixed with 4% paraformaldehyde in PBS (Nacalai Tesque, Kyoto, Japan), rinsed with 0.5% Triton X‐100 in PBS for 5 min three times, and mounted with media containing 4’,6‐diamidino‐2‐phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA). Samples were analyzed using an LSM 700 Confocal Laser Scanning Microscope (Carl Zeiss, Tokyo, Japan) equipped with a Plan‐Apochromat ×63 (numerical aperture 1.4) lens. The DAPI, Oregon‐Green, and TMR fluorescence were detected at excitation wavelengths of 405, 488, and 555 nm with 300–483, 493–550, and 560–800 nm band‐pass filters, respectively. Images were processed by using zen 2012 imaging software (Carl Zeiss).
3
Immunofluorescent Staining and Confocal Imaging
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Immunofluorescent staining was performed as described previously20 (link). We used a LSM780 laser-scanning confocal microscope (ZEISS, Germany) equipped with a Plan-Apochromat 63 × 1.4 objective for all imaging.
4
Confocal Imaging of Fluorescent Cellular Markers
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Images were acquired with a Zeiss LSM 710 inverted confocal laser scanning microscope with a Plan-Apochromat 63× and 1.4 NA M27 oil immersion objective using immersion oil (Carl Zeiss). DAPI, Alexa Fluor 488, Alexa Fluor 555, Alexa Fluor 647 and 5TYE563 were excited at 405, 488, 543, 647 and 543 nm, respectively. The zoom factor was set to 1–4×, and X- and Y-scanning sizes were each 1024 pixels. All other images except for HEK293 IF were acquired as z-stacks, and the z-scanning size was 0.979–2 μm.
5
Microscopic Feather Examination
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After treatment with 2% OsO4, chicken feathers were rinsed in distilled water and mounted under glass cover slips using Mowiol 4-88 (Merck KGaA, Darmstadt, Germany). Light micrographs were taken using an Axio Imager Z1 Zeiss widefield microscope (Carl Zeiss Microscopy GmbH, Oberkochen, Germany) equipped with a Plan-Apochromat × 63.
6
Confocal Microscopy of Tissue Samples
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Images were acquired using a 63× oil objective (Plan-Apochromat 63× [numerical aperture, 1.46] oil immersion objective for differential interference contrast [DIC]; Carl Zeiss, Jena, Germany). All sections were analyzed using a confocal laser microscopy system and software (LSM710, Carl Zeiss, Jena, Germany) that was built around an inverted microscope (Axio Observer Z1, Carl Zeiss, Jena, Germany) as previously described [40 (link)]. The images were saved in TIFF format and analyzed by ImageJ software (Wayne Rasband, National Institutes of Health, Bethesda, MD, USA).
7
Confocal Laser Scanning Microscopy Protocols
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CLSM was either performed on a Nikon C2+ confocal laser scanning microscope or on a LSM 510 (Zeiss) confocal laser scanning microscope. The Nikon C2+ microscope was equipped with four lasers: a 405 nm diode laser (100 mW, Coherent Inc.), a 488 nm DPSS Laser (10 mW, Melles Griot GmbH), a 543 nm HeNe laser (5 mW, Melles Griot GmbH), and a 642 nm diode laser (45 mW, Melles Griot GmbH). The intensity of the laserlight was controlled by an acusto optical tunable filter (AOTF). We used a 60× violet corrected oil objective with a NA of 1.4 for imaging (Plan Apo VC 60× H, Nikon). The LSM 510 microscope was equipped with three lasers: a multiline argon laser (458 nm, 477 nm, 488 nm, 514 nm, 30 mW, LASOS Lasertechnik), a 543 nm HeNe laser (1 mW, LASOS Lasertechnik) and a 633 nm HeNe laser (5 mW, LASOS Lasertechnik). The intensity of the laser light was controlled by an acusto optical tunable filter (AOTF). For imaging a 63× objective with a NA of 1.4 (Plan Apochromat 63×, Zeiss) was used.
8
Superresolved Fluorescence Microscopy Protocols
9
Live Cell Imaging with Zeiss Microscope
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Microscopic images of living cells were obtained as described by Nanbu et al. [15 (link)], using an AxioCam digital camera (Zeiss) connected to an Axio Observer.Z1 microscope (Zeiss) with a Plan-Apochromat 63× objective lens (numerical aperture, 1.4).
10
Confocal Imaging of Alexa 647 and DAPI
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All samples were mounted in Slow Fade Gold mounting media. Samples were imaged on a Zeiss LSM 510 META confocal microscope (Carl Zeiss, Germany) (Plan-Apochromat 63×/1.2 W objective). The Alexa 647 fluorophore was excited with a 633 nm laser and the DAPI with a 405 nm laser. Images were processed using Zeiss LSM Image Browser (Carl Zeiss) or ImageJ.
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