Following euthanization, left lobes were fixed with 4% paraformaldehyde, stored at 4 °C overnight for fixation of the tissue, mounted in paraffin, and 5 µm sections were affixed to glass microscope slides for histopathology as previously described [61] . Sections were prepared for immunofluorescence by deparaffinizing with xylene and rehydrating through a series of ethanols. For antigen retrieval, slides were heated for 20 min in 95 °C pH 6.0 sodium citrate buffer with 0.05% TWEEN-20 then rinsed in dH2O. Sections were then blocked for 1 h in 1% BSA in PBS, followed by incubation with primary antibody for PDIA3 (LSBio, LS-B9768) at 1:300, overnight at 4 °C. Slides were then washed 3 × 5 min in PBS, incubated with Alexafluor 647 at 1:1000 in 1% BSA, and counterstained with DAPI in PBS at 1:4000 for nuclear localization. Sections were imaged using a Zeiss 510-META confocal laser-scanning microscope. Images were captured at x40 magnification in oil immersion. The image files were converted to Tiff format. Brightness and contrast were adjusted equally in all images.
510 meta laser scanning confocal microscope
Manufactured by Zeiss
Sourced in Germany, United States
The Zeiss 510 Meta-laser scanning confocal microscope is a high-performance instrument designed for advanced imaging applications. It utilizes a multi-line laser system to provide excitation across a range of wavelengths, enabling the visualization of diverse fluorescent samples. The microscope's confocal design offers enhanced optical sectioning and improved signal-to-noise ratios, making it well-suited for a variety of imaging tasks.
Automatically generated - may contain errors
Lab products found in correlation
Eight well chambered coverglass slides, by Thermo Fisher Scientific (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions)
Live dead baclight stain, by Thermo Fisher Scientific (3 mentions)
Alexa fluor 488 conjugated goat anti mouse igg, by Cell Signaling Technology (3 mentions)
Fm4 64, by Thermo Fisher Scientific (3 mentions)
Alexa fluor 488 conjugated anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
K alpha, by Thermo Fisher Scientific (2 mentions)
Lsm 800 airyscan microscope, by Zeiss (2 mentions)
Dapi mounting media for nuclear stain, by Vector Laboratories (2 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions)
Lsm image browser, by Zeiss (2 mentions)
Alexa fluor labeled goat anti rabbit mouse igg, by Thermo Fisher Scientific (2 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (2 mentions)
Dmr microscope, by Zeiss (1 mentions)
Pdonr207 vector, by Thermo Fisher Scientific (1 mentions)
Lsm image software, by Zeiss (1 mentions)
Primary anti gfp antibody, by Abcam (1 mentions)
Glass coverslip, by Avantor (1 mentions)
Fluoromount g, by Southern Biotech (1 mentions)
Ultimate 3000 system, by Thermo Fisher Scientific (1 mentions)
74 protocols using 510 meta laser scanning confocal microscope
1
Immunofluorescent Analysis of PDIA3 Expression
2
Immunocytochemical Analysis of HEE Markers
3
Subcellular Localization of PARP1 and PARP2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The full-length cDNAs of PARP1/2 and PARG1/2 were subcloned into the pDONR 207 vector (Invitrogen) and introduced into the destination vector pGWB405, resulting in constructs with C-terminal fusions to GFP under the control of 35S promoter. The sequence-verified constructs were transformed into Agrobacterium tumefaciens strain GV3101(pMP90) and transiently expressed in Nicotiana benthamiana leaves by infiltration. Confocal fluorescence microscopy was carried out at indicated times using a Zeiss 510 Meta confocal laser scanning microscope. Agrobacterium strains carrying 35S:PARP1-GFP and 35S:PARP2-GFP were also used to transform wild-type Col-0 Arabidopsis by floral dip [80 (link)]. Stable transgenic T2 lines were selected and the subcellular locations of PARP1 and PARP2 were examined by confocal microscopy.
4
Cholesterol Manipulation in Cell Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cholesterol depletion or loading studies, cells were incubated with methyl-β-cyclodextrin (MβCD) or the cholesterol-MβCD complex in Hank’s balanced salt solution (HBSS) supplemented with 25 mM HEPES, pH 7.4. Briefly, the cell medium was removed, cells were washed twice with Tris-buffered saline (TBS) and incubated with cholesterol depletion buffer (0.5% MβCD in HBSS, 25mM HEPES, pH 7.4) or loading buffer (1 mM cholesterol in 0.5% MβCD in HBSS, 25mM HEPES, pH 7.4) at 37° for 30 min. Cells were washed once with TBS and HBSS and were imaged in HBSS, 25 mM HEPES, pH 7.4 on a Zeiss 510 META confocal laser scanning microscope (CLSM).
5
Imaging Vascularized Murine Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At 30 min before killing mice, 200 μg TRITC-labeled BS-1 lectin (Sigma) was administered intravenously to stain perfused vessels. To create frozen sections, tissues or in vivo Matrigel were immersed in optimum cutting temperature compound and then frozen at −80 °C. Frozen tissues were cut by cryotome to a thickness of 10 to 50 μm. The following antibodies were used in overnight incubations: fluorescein isothiocyanate-labeled antibody against α-smooth muscle actin (Sigma) for hindlimb muscle and TRITC-labeled antibody against α-smooth muscle actin (Sigma) for in vivo Matrigel. Tissues were imaged on a 510 META confocal laser scanning microscope (Zeiss, Jena, Germany). Samples were excited with 405, 458, 488, 514, 543 or 633 nm laser lines with a BP420-480, LP505, BP560-615 and LP650 chroma filter set or a variable spectral setting with a meta-detector using Plan-Neofluar objectives (20 × /0.5 or 40 × /1.3 Oil). Three-dimensional stacks of the tissues were taken at 2–4 μm intervals spanning whole GFP (+) cells (20–40 μm). The pinhole settings were at 2.0 airy units. The Z-stack images were reconstructed as three-dimensional images or reformatted by projection view using Zeiss LSM image software.
6
Confocal Microscopy of miRNA Immunofluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Zeiss 510 META Confocal Laser Scanning microscope equipped with Carl Zeiss Zen2009 image software was used to view and image immunofluorescence slides. For quantification of immunofluorescence intensity, four independent images from two independent experiments were obtained for each condition (Negative miR, miR-31, miR-200a, Mix).
7
Visualizing Bone Osteogenesis in Zebrafish
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin sections from Tg(sp7:EGFP)b1212 transgenic skulls at age 12 wpf (21 mm SL) were collected for immunohistochemistry. The primary anti-GFP antibody (Abcam) diluted to 1:500 and secondary Alexa Fluor 488 Donkey Anti-rabbit IgG diluted to 1:500 were used to detect the GFP reporter per manufacturer’s recommendations. DAPI counterstaining allowed for nuclei visualization. Specimens were observed using Zeiss 510 META Confocal Laser Scanning Microscope and 488 nm and 405 nm laser lines.
8
Expressing RRP42-GFP Fusion in Arabidopsis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To express the RRP42-GFP fusion protein under the control of the 35S promoter, the full-length coding sequence except the stop codon was cloned from the cDNA of WT plants using the primers: GFP-LP (5′-TCTAGAATGGGGCTTTCTCTTGGGGA-3′) and GFP-RP (5′-GGTACCAGATTCGTCTTCGCAGGCCT-3′). The PCR products were fused to the pSuper 1300-GFP vector. The GFP is tagged at its C terminus. The protoplast extraction and plasmid transformation procedures were performed according to a previously described method (Kim and Somers, 2010 (link)). We used the floral dip method to obtain the stable Arabidopsis transformants, and putative transgenic plants were screened on MS plates containing 25 mg L-1 hygromycin. We used a Zeiss 510 META confocal laser scanning microscope to observe the GFP fluorescence of the transgenic plants and the RFP and GFP fluorescence of the transgenic protoplasts. To confirm that the localization of RRP42 was unaffected by GFP fusion, the 35S:RRP42 vector was transformed into the WT plants. The 35S:RRP42 stable Arabidopsis transformants displayed no obvious phenotype with the 35S:RRP42:GFP stable Arabidopsis transformants.
9
Immunofluorescence Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were plated on glass coverslips (ø 13 mm, VWR) and incubated overnight. Cells were washed twice with PBS and fixed using 4% paraformaldehyde in PBS for 15 min. Cells were then washed twice with PBS and permeabilized for 10 min using 0.5% Triton X in PBS before being blocked with IFF (1% BSA, 1% FCS in PBS) for 1 h. Coverslips were incubated overnight with primary antibody at 4°C before being washed and incubated with fluorophore conjugated secondary antibody for 1 h. Coverslips were again washed twice with PBS and incubated in 1 μg/ml DAPI solution (Biotium) for 10 min, washed and mounted using Fluoromount G (Southern Biotech). DAPI was excited at 405 nm and emitted fluorescence (peak 465 nm) collected using 435–485 band pass filter and red fluorescence captured following excitation at 543 nm and collected using a 560 nm long pass filter. Images were acquired with a Zeiss 510 META confocal laser scanning microscope using a 63× oil immersion objective.
10
Intestinal Microvascular Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pups were anesthetized and Alexa Fluor 647-conjugated-wheat germ agglutinin (500 μl of 40 μg/ml) was infused intracardiacally as previously described50 (link). Whole intestinal tissues were collected immediately after perfusion and fixed in formalin. Intestinal microvascular network images were captured by a Zeiss 510 META confocal Laser Scanning Microscope and vessel areas were analyzed as previously described50 (link).
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!