Lentivirus transduced MDCK or transiently transfected HEK-293 T cells with EpCAM-EGFP constructs were lightly trypsinized after 48 h and plated on a glass bottom 35 mm dish with serum-free Opti-MEM media (ThermoFisher Scientific, Grand Island, New York). Cells were visualized and captured with a fluorescence microscope (EVOS digital inverted microscope at 20X or 40X magnifications). Duplicate cell culture was used for data measurements on a Zeiss LSM 880 microscope equipped with the AiryScan detector, an Argon laser (Melles-Griot) for 488 nm excitation and a Zeiss Plan-Apochromat 63 × 1.4 NA DIC M27 Oil objective. The microscope is equipped with temperature and CO2 controls that were kept at 37 °C and 5%, respectively.
Plan apochromat 63 1.4 na dic m27 oil objective
Manufactured by Zeiss
The Plan-Apochromat 63x/1.4 NA DIC M27 Oil objective is a high-performance microscope objective designed for advanced imaging applications. It features a numerical aperture of 1.4 and a magnification of 63x, providing excellent resolution and light-gathering capabilities. The objective is optimized for use with immersion oil and is compatible with differential interference contrast (DIC) microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Zen software, by Zeiss (3 mentions)
Lsm 880 microscope, by Zeiss (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Opti mem media, by Thermo Fisher Scientific (1 mentions)
Human epcam duoset elisa kit, by R&D Systems (1 mentions)
35 mm glass bottom dishes, by Ibidi (1 mentions)
Ixon 897 emccd camera, by Oxford Instruments (1 mentions)
Elyra system, by Zeiss (1 mentions)
Fugene hd, by Promega (1 mentions)
7 protocols using plan apochromat 63 1.4 na dic m27 oil objective
1
Visualizing EpCAM-EGFP Fusion Proteins
2
High-Resolution Microscopy of Live Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All measurements were performed on a Zeiss LSM 880 microscope equipped with the AiryScan detector, an Argon laser (Melles-Griot) for 488 nm excitation and a Zeiss Plan-Apochromat 63×/1.4 NA DIC M27 Oil objective. The acquisition modality was set to Spot acquisition with 2.46 µs per time point, bit depth was set at 16 bits, gain at 750 and the pinhole aperture was set to max. All files were successively saved in the ome.tiff format to be processed from our MATLAB routine. The microscope is equipped with temperature and CO2 controls, that were kept at 37 °C and 5%, respectively.
3
Chlamydomonas Airyscan Microscopy Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We prepared Chlamydomonas cells for Airyscan microscopy according to the method previously used for SIM (Iwai et al. 2018) . Briefly, the cultures grown in TAP liquid medium were centrifuged at 3000×g and 23 °C for 1 min.
The pelleted cells were resuspended with 0.5% low-melting-point agarose in TAP medium and mounted between two coverslips placed in an Attofluor cell chamber. Chlamydomonas cells were observed using a Zeiss LSM 880 microscope equipped with the Airyscan detector with a Zeiss Plan-Apochromat 63 × /1.4 NA DIC M27 Oil objective. Chls were excited with 633 nm laser, and fluorescence was acquired through a 645 nm longpass filter. Image acquisition and analysis were done under the full control of ZEN software (Zeiss) and ImageJ software (US National Institutes of Health, https:// www. nih. gov/).
The pelleted cells were resuspended with 0.5% low-melting-point agarose in TAP medium and mounted between two coverslips placed in an Attofluor cell chamber. Chlamydomonas cells were observed using a Zeiss LSM 880 microscope equipped with the Airyscan detector with a Zeiss Plan-Apochromat 63 × /1.4 NA DIC M27 Oil objective. Chls were excited with 633 nm laser, and fluorescence was acquired through a 645 nm longpass filter. Image acquisition and analysis were done under the full control of ZEN software (Zeiss) and ImageJ software (US National Institutes of Health, https:// www. nih. gov/).
4
Visualizing EpCAM Expression in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Retrovirus transduced MDCK or transiently transfected HEK-293T cells with EpCAM-GFP constructs were lightly trypsinized after 48 hours and plated on a glass bottom 35 mm dish with serum-free Opti-MEM media (ThermoFisher Scienti c, Grand Island, New York). Cells were visualized and captured with a uorescence microscope (EVOS digital inverted microscope at 20X or 40X magni cations). Duplicate cell culture was used for data measurements on a Zeiss LSM 880 microscope equipped with the AiryScan detector, an Argon laser (Melles-Griot) for 488 nm excitation and a Zeiss Plan-Apochromat 63×/1.4 NA DIC M27 Oil objective. The microscope is equipped with temperature and CO 2 controls that were kept at 37 °C and 5%, respectively.
EpCAM ELISA HEK-293T transiently transfected cells or tumor cell lines were plated in serum free Opti-MEM media.
After 48 hours, conditioned media was used as a source of secreted/soluble EpCAM. Human EpCAM DuoSet ELISA kit (DY960) was used from R&D Systems Inc. (Minneapolis, MN) as recommended.
EpCAM ELISA HEK-293T transiently transfected cells or tumor cell lines were plated in serum free Opti-MEM media.
After 48 hours, conditioned media was used as a source of secreted/soluble EpCAM. Human EpCAM DuoSet ELISA kit (DY960) was used from R&D Systems Inc. (Minneapolis, MN) as recommended.
5
Visualizing Chlorophyll Fluorescence in C. reinhardtii
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C. reinhardtii cells were observed using a Zeiss LSM 880 microscope equipped with the Airyscan detector with a Zeiss Plan-Apochromat 63×/1.4 NA DIC M27 Oil objective. Chls were excited with 633 nm laser, and fluorescence was acquired through a 645 nm longpass filter. Image acquisition and analysis were done under the full control of ZEN software (Zeiss) and ImageJ software (US National Institutes of Health, https://www.nih.gov/).
6
Live Cell Imaging Microscopy Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For live imaging, cells were plated onto 35 mm glass bottom dishes (ibidi GmbH, Germany) and transiently transfected using Fugene HD (Promega, USA) 24 hr prior to imaging. All cultures were maintained at 37°C and 5% CO2 for the duration of the experiment. Confocal microscopy was performed on a Zeiss 880 equipped with a Plan-Apochromat 63×/1.4 NA Oil DIC M27 objective and an Airyscan detector. TIRF imaging was performed using the Zeiss Elyra system fitted with a Plan-Apochromat 100×/1.46 NA Oil objective and an Andor iXon 897 EMCCD camera. In both cases, samples were illuminated using 488 nm, 561 nm or 633 nm lasers, and all data was collected using the Zen software (Zeiss).
7
C. elegans Colocalization of ABCB6 and HMT-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transgenic strains were grown in normal growth conditions at 20 °C. To test the subcellular co-localization of ABCB6 and CeHMT-1, the phmt-1::ABCB6::mCherry (TTV677) strain was crossed with phmt-1::HMT-1::GFP (VF31) males and the F1 progeny co-expressing both transgenes was examined with a confocal microscope (Zeiss LSM 710, Plan-Apochromat 63 ×/1.4 NA Oil DIC M27objective). Lysosomal staining was performed as described in [24 (link)]. To determine the subcellular localization of CeHMT-1::GFP and ABCB6::GFP, phmt-1::hmt-1::gfp (VF31) and phmt-1::ABCB6::gfp (TTV634) were crossed with strains expressing different endosomal markers [25 (link)], resulting in TTV701 unc-119(ed3)III; eluIs310[phmt-1::ABCB6::gfp + unc-119(+)]; qxIs110(Pges-1mCHERRY::RAB-5), TTV702 unc-119(ed3)III; eluIs310[phmt-1::ABCB6::gfp + unc-119(+)]; qxIs111(Pges-1mCHERRY::RAB-7), TTV703 unc-119(ed3)III; eluIs310[phmt-1::ABCB6::gfp + unc-119(+)]; qxIs213(Pges-1mCHERRY::RAB-10), TTV705 unc-119(ed3)III; gfIs1[phmt-1::hmt-1::gfp, unc-119(+)]; qxIs110(Pges-1mCHERRY::RAB-5), TTV706 unc-119(ed3)III; gfIs1[phmt-1::hmt-1::gfp, unc-119(+)]; qxIs111(Pges-1mCHERRY::RAB-7), TTV707 unc-119(ed3)III; gfIs1[phmt-1::hmt-1::gfp, unc-119(+)]; qxIs213(Pges-1mCHERRY::RAB-10).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!