Confocal fluorescence-microscopy to visualize tubulin aggregates, multinucleation, Annexin V-FITC/PI or Alexa Fluor 594 TUNEL-staining in HTLV-1 Tax-expressing cells was performed on a Zeiss LSM800 instrument with an Airyscan super-resolution detector and stage CO2 incubator (Carl Zeiss Microscopy). All images were taken using either Plan-Apochromat 40x/1.3 or Plan-Apochromat 63x/1.4 oil immersion objectives and Zeiss ZEN system software. The expression of HTLV-1 p30II-GFP in cotransfected cells was visualized on an inverted Nikon Eclipse TE2000-U microscope and D-Eclipse C1 confocal system (Nikon Instruments, Melville, NY) equipped with 633 nm and 543 nm He/Ne and 488 nm Ar lasers using a Plan Apo 20x/0.75 objective lens.
Plan apochromat 40x 1
Manufactured by Zeiss
Sourced in Germany
The Plan-Apochromat 40x/1.3 is a high-performance objective lens from Zeiss. It is designed to provide a flat, distortion-free field of view with excellent chromatic correction. The numerical aperture of 1.3 allows for high-resolution imaging and light collection.
Automatically generated - may contain errors
Lab products found in correlation
Plan apochromat 63x 1, by Zeiss (3 mentions)
Plan apochromat 20x 0, by Zeiss (3 mentions)
Zen system software, by Zeiss (2 mentions)
Axiocam mrm, by Zeiss (2 mentions)
Zen 2, by Zeiss (2 mentions)
Lsm810 confocal microscope, by Zeiss (2 mentions)
Plan neofluar 10x 0, by Zeiss (2 mentions)
Airyscan super resolution detector, by Zeiss (1 mentions)
D eclipse c1 confocal system, by Nikon (1 mentions)
Eclipse te2000 u microscope, by Nikon (1 mentions)
Lsm 800, by Zeiss (1 mentions)
Axiovision module z stack, by Zeiss (1 mentions)
Hcx pl apo cs 40x 1, by Leica (1 mentions)
Axio observer z1 microscope, by Zeiss (1 mentions)
Axiovision 4, by Zeiss (1 mentions)
Lsm 800 instrument, by Zeiss (1 mentions)
Axio imager z2 microscope, by Zeiss (1 mentions)
Prolong gold, by Thermo Fisher Scientific (1 mentions)
Axio observer, by Zeiss (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
10 protocols using plan apochromat 40x 1
1
Visualizing HTLV-1 Tax-induced Cellular Stress
2
Time-Lapse Microscopy Imaging of Tumor Spheres
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Time-lapse microscopy imaging was performed on a Zeiss AxioObserver Z1 microscope (Carl Zeiss, Germany) with a Plan-Apochromat 40X/1.3 (NA = 1.3, working distance = 0.21 mm) or Plan-Apochromat 63X/1.4 (NA = 1.4, working distance = 0.19mm) oil lens objective, a camera (AxioCam MRm; Carl Zeiss), and Apotome (ApoTome 2; Carl Zeiss). z-stack images were taken with AxioVision module Z-stack (Zeiss). Three-dimensional reconstruction of z-stacks and two-dimensional projections were conducted using the Imaris 7.1 Software (Bitplane Scientific Software, Belfast, UK) and the Z Projection ImageJ plugin, respectively, 35. For confocal reflection microscopy, tumor-spheres were seeded on glass bottom 35mm-μ-dishes, incubated for different times and subsequently fixed with 4% formalin. Z-stack images were acquired using a Leica TCS-SP2 AOBS confocal microscopy with a HCX PL APO CS 40x/1.25 oil lens objective and a 488 nm Argon laser in reflection at room temperature.
3
Fluorescence Microscopy Imaging of Cellular Assays
4
Immunostaining of Adherent Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were plated on gelatin-coated glass coverslips. Following agonist stimulation, coverslips were washed twice in PBS and fixed with 2% (w/v) PFA for 15 min at room temperature. Cells were quenched in PBS containing 50 mM ammonium chloride and then permeabilised in PBS containing 0.2% (w/v) Triton X-100 for 10 min at room temperature. Cells were washed in PBS and then blocked for 1 h at room temperature with TBS containing 0.1% (v/v) Tween-20 and 5% (w/v) BSA. Coverslips were incubated with primary antibody in TBS containing 0.1% (v/v) Tween-20 and 1% (w/v) BSA. Cells were washed × 5 in TBST and then incubated with the relevant secondary Alexa Fluor coupled antibodies for 1 h at room temperature. Samples were then incubated for 10 min with Hoechst 33342 (Invitrogen #H-21492) and mounted with Prolong gold (Invitrogen). Fluorescent images were acquired with a Zeiss AxioObserver inverted fluorescence microscope with Apotome2 using a Plan-Apochromat 40x/1.3 oil immersion objective and Zeiss ZenPro software.
5
Microscopy Imaging of BCT Probes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microscopy imaging was performed on a Cell Observer equipment composed of a Zeiss AxioObserver Z1 wide field inverted fluorescence microscope (Carl Zeiss, Germany) with a Plan-Apochromat 40X/1.3 (NA = 1.3, working distance = 0.21 mm) or Plan-Apochromat 63X/1.4 (NA = 1.4, working distance = 0.19 mm) oil lens objective, a camera (AxioCam MRm; Carl Zeiss), a CO2 incubator, and a ZEISS ApoTome.2 structured illumination system. ApoTome.2 allows acquisition of optical sections of the fluorescent sample by moving the appropriate grid into the beam path and calculating the optical section from three images with different grid positions without time lag. Acquisition and processing of images were conducted using the Zen (Carl Zeiss) software. For direct comparison of fluorescence intensities between the two BCT probes (Fig. 2 ), microscope settings were the followings: 420 ms exposure time with a 0.79 μm depth in focus, 1.17 μm section thickness and maximum intensity of the HXP 120 C lighting unit. Images in Fig. 3–5 were acquired with the maximum intensity of the HXP 120 C lighting unit and exposure times of 420 ms for (R)-BCT-2 and 20 ms for MitoTracker.
6
Fluorescence Microscopy Analysis of mKeima-LC3B
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescence microscopy images were acquired with an Axio Observer Z1 platform with a Plan-Apochromat 40X/1.3, (working distance, 0,21 mm) equipped with an ApoTome.2 system and an Axiocam MRm camera (from Carl Zeiss, Jena, Germany). Zeiss Immersol® immersion oil was used for all microscopic analyses. mKeima-LC3B ratio determination and colocalization of mKeima-LC3B (Ex586/Ex440) were measured with Spots colocalization (ComDet) ImageJ plugin.
7
Live-Cell Imaging of Nitric Oxide Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Live-cell imaging experiments were performed on an inverted wide-field epi-fluorescence microscope Zeiss Axio Observer.Z1/7 (Carl Zeiss AG, Oberkochen, Germany) equipped with an LED light source Colibri 7 (423/44 nm, 469/38 nm, 555/30), Plan-Apochromat 20x/0.8 dry objective, Plan-Apochromat 40x/1.4 oil immersion objective, a monochrome C.C.D. camera Axiocam 503, and a custom-made gravity-based perfusion system. O-geNOps signals were imaged by exciting cells (555 nm) using a motorized dual filter wheel equipped with the filter combinations FT570 (BS) and emission filter 605/70. Control and data acquisition were executed using Zen Blue 3.1 Pro software (Carl Zeiss AG, Oberkochen, Germany). Administration and withdrawal of 3-(2-Hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine (NOC-7) were performed using a custom-made perfusion system connected to a metal perfusion chamber (NGFI, Graz, Austria).
8
Confocal Microscopy Imaging Workflow
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Images were acquired using a LSM810 confocal microscope (Zeiss) equipped with an EC Plan-Neofluar 10 x/0.3, a Plan-Apochromat 20 x/0.8 or a Plan-Apochromat 40 x/1.4 Oil DIC objective. Images were taken using Zen2.1 software (Zeiss) and processed and quantified with Fiji (NIH). Figures were assembled using Affinity Photo and Windows Office Powerpoint.
9
Visualizing phosphorylated and total DP in cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells plated on glass coverslips were fixed either in anhydrous ice-cold methanol for 3 min on ice to visualize phospho S2849 DP/total DP staining or 4% paraformaldehyde (PFA) solution for 20 min at room temperature followed by anhydrous ice-cold methanol for 3 min on ice to visualize B55α. Cells were blocked with either 5% goat serum or 1% BSA and 1% donkey serum. All samples were mounted onto glass slides with ProLong Gold antifade reagent (Thermo Fisher Scientific). 3D reconstructed skin and human skin samples were stained from frozen embedded samples and fixed as described above. For PLA analysis, samples were fixed as described above and PLA was performed as described in Hegazy et. al29 (link).
Apotome images were acquired using ZEN 2.3 software with an epifluorescence microscope system (Axio Imager Z2, Carl Zeiss) fitted with an X-Cite 120 LED Boost System, an Apotome.2 slide module, Axiocam 503 Mono digital camera, and a Plan-Apochromat 40x/1.4, Plan-Apochromat 63x/1.4 objective, Plan-Apochromat 100x/1.4 objective (Carl Zeiss). Images are processed using ImageJ software. Colocalization analysis was performed using the JaCoP ImageJ plugin28 (link).
Apotome images were acquired using ZEN 2.3 software with an epifluorescence microscope system (Axio Imager Z2, Carl Zeiss) fitted with an X-Cite 120 LED Boost System, an Apotome.2 slide module, Axiocam 503 Mono digital camera, and a Plan-Apochromat 40x/1.4, Plan-Apochromat 63x/1.4 objective, Plan-Apochromat 100x/1.4 objective (Carl Zeiss). Images are processed using ImageJ software. Colocalization analysis was performed using the JaCoP ImageJ plugin28 (link).
10
Confocal Microscopy Imaging Workflow
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Images were acquired using a LSM810 confocal microscope (Zeiss) equipped with an EC Plan-Neofluar 10x/0.3, a Plan-Apochromat 20x/0.8 or a Plan-Apochromat 40x/1.4 Oil DIC objective.
Images were taken using Zen2.1 software (Zeiss) and processed and quantified with Fiji (NIH). Figures were assembled using Adobe Affinity Photo and Windows Office Powerpoint.
Images were taken using Zen2.1 software (Zeiss) and processed and quantified with Fiji (NIH). Figures were assembled using Adobe Affinity Photo and Windows Office Powerpoint.
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!