For laser ablation experiments on SRSF2-GFP droplets, mouse oocytes were imaged at 37 °C with a Zeiss (Axio Observer.Z1/7) LSM 980 confocal microscope equipped with Airyscan 2, 2 PhotoMultiplier Tubes (PMT), a GaAsP spectral sensor, a Plan-APO 40x/1.3 oil immersion objective, a 488 nm 10 mW laser, and a motorized deck with a temperature-controlled chamber. Images were acquired using Zen 3.0 software. A total of 150 frames of 579 × 579 pixels (53 µm × 53 µm; 16-bit depth) were acquired on single z-planes with a bidirectional scan speed of 6 and 1.43 s intervals for a total of 214 s. Three frames preceded the bleaching (full 488 nm 10 mW laser power) of a fixed 8 µm × 2 µm region of SRSF2-GFP droplet-containing nucleoplasm. The bleached region size was fixed to be larger than the target droplet and thus included the neighboring dissolved SRSF2-GFP phase. Fluorescence recovery was imaged with 1.2% laser power.
Axio observer z1 7
Manufactured by Zeiss
Sourced in Germany
The Axio Observer.Z1/7 is a high-performance inverted microscope system designed for advanced imaging applications. It features a stable and robust construction, providing a solid platform for various imaging techniques. The microscope is equipped with a range of optical components and accessories to enable diverse imaging modalities, such as brightfield, phase contrast, and fluorescence microscopy.
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Lab products found in correlation
Axio scope a1, by Zeiss (3 mentions)
Fluorsave reagent, by Merck Group (2 mentions)
Ms 222, by Merck Group (2 mentions)
880 confocal microscope, by Zeiss (2 mentions)
Axiocam 512 mono, by Zeiss (2 mentions)
Orca flash 4.0 lt cmos digital camera, by Hamamatsu Photonics (2 mentions)
Na 1.1 ld c apochromatic water immersion objective, by Hamamatsu Photonics (2 mentions)
Alpha plan apochromat 100 1.46 oil dic, by Zeiss (1 mentions)
Mitosox red, by Thermo Fisher Scientific (1 mentions)
Plan apochromat 20 0.8 na objective, by Zeiss (1 mentions)
Lsm 900 confocal, by Zeiss (1 mentions)
Matlab, by MathWorks (1 mentions)
Lsm 800 microscope, by Zeiss (1 mentions)
Fm4 64 dye, by Thermo Fisher Scientific (1 mentions)
Zen 3.2 blue edition software, by Zeiss (1 mentions)
Fluorescence mounting medium, by Agilent Technologies (1 mentions)
Fluoview 1000 laser scanning microscope, by Olympus (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 633 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Dako fluorescence mounting medium, by Agilent Technologies (1 mentions)
26 protocols using axio observer z1 7
1
Laser Ablation of SRSF2-GFP Droplets
2
Fluorescence Microscopy of Cells
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Fluorescence microscopy was performed on a Zeiss Axio Observer Z1/7 (Carl Zeiss, Germany) inverted wide-field microscope, equipped with a Colibri 7 LED light source, an Alpha Plan-Apochromat × 100/1.46 oil DIC (UV) M27 objective, filter sets 38 HE (ex. 450–490, dichroic beam splitter 495, em. 500–550; sfGFP), 43 HE (ex. 538–562, beam splitter 570, em. 570–640; mCherry) and an Axiocam 506 Mono camera. Samples were embedded onto 1% agarose pads on metal microscopy slides prior to imaging. Cell numbers were counted manually from the images.
3
Immunofluorescence Staining of Cytokine-Treated Cells
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Cells were seeded on coverslips at 2.5 × 105 cells/well and then treated with required concentrations of cytokines. Treated cells were then washed with 1X PBS and fixed with 4% paraformaldehyde, followed by incubation at room temperature for 10–15 min. After that, cells were treated with 0.2% Triton X for approximately 2 min. Post multiple PBS washes, cells were blocked with 2.5% bovine serum albumin (BSA) blocking solution for 1 h. The cells were then incubated overnight with primary antibody (1:500 dilution in 2.5% BSA) at 4°C, followed by washing twice with PBS, and then incubated with TRITC-conjugated secondary antibody (1:1,000 in 2.5% BSA) for 1 h. Coverslips were mounted on slides using antifade DAPI and imaged under a fluorescent microscope (Zeiss, Axio Scope A1, or Axio Observer.Z1/7). Images were analyzed using the Zen 3.2 (blue edition) software (48 (link)).
4
Electroporation and Imaging of Sea Urchin Embryos
5
Quantifying Mitochondrial Superoxide via MitoSOX
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The amount of Superoxide formed by mitochondria of certain cells can be determined by MitoSOX Red (Invitrogen). To check this Superoxide produced by the mitochondria of the cells, the MCF7 cells were seeded in a 6-well plate over a coverslip, and upon gaining their morphology, the cells were subjected to treatment for 1 hr, 3 hr, and 6 hr, respectively. After the drug incubation, 1 mL of 5 µM MitoSOX was added to each well and incubated for 10 min at room temperature. The cells were then washed with warm buffer thrice and fixed using Paraformaldehyde, and finally, the coverslips were mounted with the anti-fading agent, DAPI, and the cells were visualized under a fluorescent microscope (Zeiss, Axio Scope A1, or Axio Observer. Z1/7). ZEN 3.2 (blue edition) software was used to analyse the images.
6
Confocal Microscopy Analysis of Coated Wood
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One beech wood sample spray coated with EOL-TC was investigated using a Zeiss LSM 900 confocal laser scanning microscope equipped (Oberkochen, Germany)on Axio Observer Z1/7 inverted stand, with diode lasers with 405 nm, 488 nm, 561 nm, and 640 nm wavelengths, and using a plan-apochromat 20×/0.8NA objective (Carl Zeiss S.p.A., Milan, Italy). The measurements were conducted in a measurement area of 319.45 × 319.45 µm using fluorescence excitation for the specific illumination of lignin (553 nm) and carbohydrate (401 nm) moieties on the coated wood sample surface. The emission wavelengths were 568 nm for lignin and 422 nm for cellulose.
7
Imaging SOD1 Aggregation Kinetics
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Purified
SOD1(A4V)-HaloTag (25 μM),P4 (5 μM), and AgHalo 550 (5 μM) were incubated at room temperature
in tris·HCl buffer (50 mM tris·HCl, 100 mM NaCl, pH 7.5)
for 20 min. EDTA (80 mM) was added, and the solution was incubated
at 25 or 59 °C for 30 min. The solution was transferred to a
glass slide equipped with a rubber spacer and topped with a glass
coverslip. Imaging was completed using a Zeiss Axio Observer.Z1/7
and an EC Plan-Neofluar 10x/0.30 Ph 1 objective.P4 fluorescence was obtained using a 630 nm LED light source,
collecting emission through a band-pass filter from 659 to 759 nm.AgHalo 550 fluorescence was obtained using a 430 nm LED light
source, collecting emission through a band-pass filter from 546 to
564 nm. Nonfluorescent images were obtained using differential interference
contrast (DIC) microscopy.
SOD1(A4V)-HaloTag (25 μM),
in tris·HCl buffer (50 mM tris·HCl, 100 mM NaCl, pH 7.5)
for 20 min. EDTA (80 mM) was added, and the solution was incubated
at 25 or 59 °C for 30 min. The solution was transferred to a
glass slide equipped with a rubber spacer and topped with a glass
coverslip. Imaging was completed using a Zeiss Axio Observer.Z1/7
and an EC Plan-Neofluar 10x/0.30 Ph 1 objective.
collecting emission through a band-pass filter from 659 to 759 nm.
source, collecting emission through a band-pass filter from 546 to
564 nm. Nonfluorescent images were obtained using differential interference
contrast (DIC) microscopy.
8
Autofluorescence Imaging of FFPE Tissue
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The autofluorescence of protein-bound NADH and oxidized flavoproteins containing flavin adenine dinucleotide (Fp) of formalin-fixed-paraffin-embedded (FFPE) tissue slides (8 µm) was imaged and quantified, as previously described [47 (link)]. NADH signals were collected using bandpass filters with excitation (Ex) at 370–400 nm, and emission (Em) at 414–450 nm. The Fp signals were acquired with Ex: 450–488 nm and Em: 500–530 nm bandpass filters using a Zeiss wide-field microscope (Axio Observer.Z1/7, One North Broadway, White Plains, NY, 10601, USA). Tile images (pixel size 1.172 × 1.172 µm2) were taken using a 5×/0.16 NA objective with shading correction on the fly, followed by photostitching. The acquired images were processed with a customized Matlab® (The MathWorks, Inc., 1 Apple Hill Dr, Natick, MA 01760, USA) routine to generate the redox images where the redox ratio Fp/(NADH + Fp) image was generated pixel-by-pixel from the NADH and Fp images and to obtain the mean value of each of the redox indices (NADH, Fp, the redox ratio) using global averaging [47 (link),48 (link),49 (link)]. Obvious fluorescent artifacts (in either channel) were removed during the imaging analysis.
9
Fluorescence Microscopy of Sporulating Cells
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Cells harboring the gfp reporter were collected at hours 5 or 6 after the onset of sporulation in DSM medium by centrifugation at 8000 rpm for 1 min. Subsequently, cells were resuspended in 0.2 mL Hanks’ balanced salt solution (HBSS) with 5 μg/mL FM4–64 dye (Molecular Probes) to stain membranes, and then spotted on thin 1% agarose pads with coverslips. Fluorescence microscopy was performed using a Zeiss LSM 800 microscope equipped with Axio Observer. Z1/7 and Plan-Apochromat 63x/1.40 Oil DIC M27 objective. Images were visualized using ZEN 3.2 (blue edition) software (Carl Zeiss, Oberkochen, Germany).
10
Localization of UTX-TagGFP2 in Stably Transduced Cells
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Analysis of UTX-TagGFP2 localization was performed in stably transduced UCCs. As previously described [33 (link)], after fixation with 4% formaldehyde, cells were permeabilized using 0.2% Triton X100 in PBS for 10 min at RT, blocked with 1% BSA in PBS, for 30 min at RT and subsequently incubated for 1 h at RT with 14 nM Rhodamine Phalloidin in blocking solution. Following counter-staining of nuclei with 1 µg/mL DAPI (4´,6-diamidino-2-phenylindole) cells were mounted with fluorescence mounting medium (DAKO, Glostrup, Denmark). Imaging was performed using ZEISS Axio Observer.Z1 / 7; Plan-Apochromat 40x/1.4 Oil DIC (UV) VIS-IR M27; 90 HE DAPI/ GFP/ Cy3 /Cy5; LED-module "wavelength" nm (Colibri 7); Axiocam 512 mono (ZEISS, Jena, Germany)
Higher resolution images were obtained using an inverted confocal FluoView1000 laser scanning microscope with a 60xW UPLSAPO objective NA1.2 in a sequential DAPI-GFP-Phalloidin scanning mode (Olympus, Hamburg, Germany). Resolution was 1024 × 1024.
Higher resolution images were obtained using an inverted confocal FluoView1000 laser scanning microscope with a 60xW UPLSAPO objective NA1.2 in a sequential DAPI-GFP-Phalloidin scanning mode (Olympus, Hamburg, Germany). Resolution was 1024 × 1024.
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