Embryos were prepared as described previously (Kong et al., 2019 (link)). Briefly, the staged embryos were collected and dechorionated with 50% hypochlorite bleach for 90 s, aligned on an agar block, and attached on the coverslips by homemade glue covered with halocarbon oil. Cross-sectional images were recorded from the dorsal side for E-CadGFP (488 nm excitation) on a spinning disc microscope with a 40x or ×100 oil objective (Carl Zeiss, ×40/oil, NA1.2, 100x/oil, NA1.4) with an emCCD camera (Carl Zeiss, AxioCam MRm). The apical plane of the embryo was acquired with axial sections of each 0.5 µm and a frame rate of 0.2/min or 1/min. Images in Figure 6A were acquired on a laser scanning confocal microscopy (Carl Zeiss, ZEISS LSM 980 with Airyscan 2) with a ×63 oil objective (Carl Zeiss, ×63/oil, NA1.4), and the apical plane of the embryo was acquired with axial sections of each 0.5 µm. Images in Figure 7A were acquired on a laser scanning confocal microscopy (Carl Zeiss, ZEISS LSM 980 with Airyscan 2) with a ×63 oil objective (Carl Zeiss, ×63/oil, NA1.4), and the apical plane of the embryo was acquired with axial sections of each 1 µm and a frame rate of 0.2/min.
Lsm 980
Manufactured by Zeiss
Sourced in Germany, United States, Japan, China
The LSM 980 is a confocal laser scanning microscope designed and manufactured by Zeiss. Its core function is to provide high-resolution imaging capabilities for scientific research and analysis, allowing users to capture detailed images of microscopic samples.
Automatically generated - may contain errors
Lab products found in correlation
Airyscan 2, by Zeiss (42 mentions)
Lsm 880, by Zeiss (39 mentions)
Lsm 700, by Zeiss (19 mentions)
Zen blue software, by Zeiss (14 mentions)
Lsm 780, by Zeiss (14 mentions)
Hoechst 33342, by Thermo Fisher Scientific (13 mentions)
Lsm 710, by Zeiss (12 mentions)
Airyscan 2 confocal microscope, by Zeiss (11 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (11 mentions)
Lsm 800, by Zeiss (9 mentions)
Axio observer z1, by Zeiss (9 mentions)
Bovine serum albumin (bsa), by Merck Group (8 mentions)
Triton x 100, by Merck Group (7 mentions)
Zen software, by Zeiss (6 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (6 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (6 mentions)
Propidium iodide, by Thermo Fisher Scientific (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (5 mentions)
Airyscan, by Zeiss (4 mentions)
Axio imager m2, by Zeiss (4 mentions)
444 protocols using lsm 980
1
Imaging Embryo Development Dynamics
2
Live and Fixed Oocyte Imaging
3
Multimodal Imaging of Retinal Cells
4
Tracing Hematopoietic Progenitor Dynamics
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Tg(fli1a1:gal4ffubs4) were outcrossed with Tg(UAS:Kaederk8) and injected with either control or miR-128 morpholino at a concentration of 0. 75 ng μl−1 at the one cell stage. Embryos were screened at 24 hpf for Kaede positive, and the ventral wall of the dorsal aorta was photoconverted with a Zeiss LSM 980 scanning confocal using a 406 nm laser at an intensity of 16% for 10 s at 30 hpf followed by incubation at 28 °C. At 4.5 dpf the thymus and CHT of photoconverted embryos were imaged with a Zeiss LSM 980 confocal. Red-Kaede-positive cells representing erythroid and myeloid progenitors were counted in the CHT and red-Kaede-positive cells representing lymphoid progenitors were counted in the thymus. Red-Kaede volume in the thymus was measured using IMARIS software (V.9.9.1, Bitplane) by utilizing the surface module to create a 3D reconstruction. Red-Kaede cells in the CHT were counted using ImageJ.
5
Autophagy Visualization Techniques
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Cells were grown in six-well plates and treated with PD2 for 24 h. Next, cells were transfected with the plasmid EGFP-LC3. After 48 h, cells were fixed with 4% paraformaldehyde for 15 min, and changes in the green fluorescence of LC3 were observed under a confocal microscope (CARL ZEISS LSM980, Germany).
The cells were grown in six-well plates and treated with PD2 for 24 h. Next, the cells were infected with mRFP-GFP-LC3 double labeled adenovirus (Cat#HB-AP2100001, Hanbio Biotechnology, China) at a dose of 20 MOI. After 48 h, the cells were fixed with 4% paraformaldehyde for 15 min, and changes in the green fluorescence of LC3 were observed under a confocal microscope (CARL ZEISS LSM980, Germany).
The cells were grown in six-well plates and treated with PD2 for 48 h. The cells were collected and centrifuged, and then subjected to fixation, dehydration, embedding, slicing, and staining. Autolysosome formation was observed using a transmission electron microscope (JEOL 1400, Japan).
The cells were grown in six-well plates and treated with PD2 for 24 h. Next, the cells were infected with mRFP-GFP-LC3 double labeled adenovirus (Cat#HB-AP2100001, Hanbio Biotechnology, China) at a dose of 20 MOI. After 48 h, the cells were fixed with 4% paraformaldehyde for 15 min, and changes in the green fluorescence of LC3 were observed under a confocal microscope (CARL ZEISS LSM980, Germany).
The cells were grown in six-well plates and treated with PD2 for 48 h. The cells were collected and centrifuged, and then subjected to fixation, dehydration, embedding, slicing, and staining. Autolysosome formation was observed using a transmission electron microscope (JEOL 1400, Japan).
6
Subcellular Localization of ZmPP2C15
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A fusion expression vector of ZmPP2C15-pMDC83 green fluorescent protein (GFP) was constructed. Protoplasts were extracted using yellowing seedlings of maize B73, and the recombinant plasmids were introduced into the protoplasts using the PEG4000 method and then cultured in the dark at 28 °C for 12–18 h [42 ]. The subcellular localization was observed using a laser confocal microscope (Zeiss LSM980, Oberkochen, Germany). A transient expression vector, ZmPP2C15-HBT, was constructed. We used the gene gun method to bombard onion epidermal cells [43 ]. The bombarded onion epidermis was cultured in hypertonic medium for about 12 h and then transferred to MS medium and cultured at 28 °C for 12 h. Subcellular localization was observed using a laser confocal microscope (Zeiss LSM980).
7
Protein-Protein Interaction Visualization
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The CDS of ABI5 and CycC1;1 or CycC1;2 were cloned into the pSPYNE or pSPYCE vector containing the Nterminal of YFP (nYFP) and or the C-terminal of YFP (cYFP), respectively. nYFP-ABI5 and cYFP-CycC1;1 or cYFP-CycC1;2 were co-expressed in N. benthamiana leaves. YFP fluorescence (excitation 488 nm, emission 543 nm) was detected under a Zeiss LSM980 laser scanning confocal microscope. H2B-mCherry (Rosa et al., 2014) was used as a nuclear marker, and the red fluorescence (excitation 561 nm, emission 600-630 nm) was also detected by Zeiss LSM980 laser scanning confocal microscope. Primer sequences are listed in Supplemental Table S1.
8
Visualizing Cytoskeleton and Nuclei in GNP-Treated Cells
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Cells were plated on 35 mm coverslip bottom dishes (MatTek, Ashland, MA USA). Following a 24 h incubation, cells cultures were dosed at a concentration of 7.5 µg/mL of functionalized Cy5-labeled GNP complex, as well as DTX at a concentration of 2.72 nM. Cell cultures were then incubated for 24 h. NucBlue™ Live ReadyProbes™ Reagent (R37605; ThermoFisher Scientific, Waltham, MA, USA) containing Hoechst 33,342 dye and CellLight™ Tubulin–GFP BacMam 2.0 (ThermoFisher Scientific, Waltham, MA, USA) was used to stain nuclei and microtubules, respectively, prior to imaging. Images were taken using a 60 × oil immersion objective lens using a confocal laser scanning microscope (Zeiss LSM 980, Carl ZeissMicroscopy GmbH, Jena, Germany).
9
Whole-Mount Immunofluorescence of Cholangiocytes
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ECO were grown in 30-μl matrigel domes in 35 mm glass bottom Petri dishes (GBD00004-200; Cell E&G LLC, San Diego, CA). Whole mount immunofluorescence for cytokeratin 7 (KRT7) and SRY-Box transcription factor 9 (SOX9), both cholangiocyte markers, was performed as previously described.[39 (link)] Mouse monoclonal anti-KRT7 antibody (#sc-400628, Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit monoclonal anti-SOX9 antibody (D8G8H, #82630, Cell Signaling, Danvers, MA) were diluted 1:100. Secondary antibodies, goat anti-mouse IgG AlexaFluor488 and chicken anti-rabbit IgG AlexaFluor594 (Invitrogen/Thermo Fisher Scientific) were used for KRT7 and SOX9, respectively, at a 1:200 dilution. 4′,6-diamidino-2-phenylindole (DAPI, 300 nM) was added together with the secondary antibodies to visualize the nuclei. The slides were mounted with ProLong™ Gold Antifade Mountant (Thermo Fisher Scientific) and analyzed by confocal microscopy using ZEISS LSM 980 (Zeiss, Oberkochen, Germany).
10
SiMYB19 Gene Expression in Rice
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The SiMYB19 coding region was amplified by PCR. The PCR product was then digested and ligated into the binary vector pCAMBIA1390 driven by a ubiquitin promoter, and the plasmid pCAMBIA1390-SiMYB19 was obtained. The latter was transformed into Oryza sativa cv. Kitaake via Agrobacterium [50 ]. The SiMYB19-16318hGFP fusion protein expression vector was constructed by inserting full-length SiMYB19 cDNA into the BamHI restriction site of the 16318hGFP vector [51 (link)]. The SiMYB19-16318hGFP fusion protein expression vector was transiently transformed via PEG into foxtail millet protoplasts isolated from plants at the two-leaf and one-heart stages. The 16318hGFP empty vector was the negative control. Green fluorescence was observed under a confocal laser scanning microscope (Zeiss LSM980; Carl Zeiss AG, Oberkochen, Germany) at ×40. The image was captured with ZEN software (Carl Zeiss AG). The PCR primers are listed in Table S1 .
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