The mNSCs were passaged every 4 days and on the 3rd day after passage, neurospheres were seeded into PLO-coated six-well plates and cultured till attachment. GO NPs at a concentration of 20 μg/mL were then added and incubated for 24 h. The morphology and migration distances of the samples were observed under a bright field of photon microscope (Nikon ECLIPSE Ti) and confocal microscope (LSM 700, Carl Zeiss, Jena, Germany).
Eclipse ti
Manufactured by Zeiss
Sourced in Germany, Japan
The Eclipse Ti is a high-performance inverted research microscope from Zeiss. It features a sturdy and stable design, with a precision motorized stage and optics optimized for advanced imaging applications. The Eclipse Ti is capable of a wide range of advanced imaging techniques, including brightfield, phase contrast, and fluorescence imaging.
Automatically generated - may contain errors
Lab products found in correlation
Axio scope a1, by Zeiss (4 mentions)
Lsm 700, by Zeiss (2 mentions)
Sim e a1r, by Nikon (2 mentions)
Eclipse e600, by Nikon (2 mentions)
Alexa fluor 488 anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Axio observer z1, by Zeiss (1 mentions)
Alexa 555 conjugated goat anti mouse antibody, by Thermo Fisher Scientific (1 mentions)
Lsm 510 meta, by Zeiss (1 mentions)
Lsm 710, by Zeiss (1 mentions)
Dcfh da, by Thermo Fisher Scientific (1 mentions)
Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (1 mentions)
Primary antibody against type 1 collagen, by Abcam (1 mentions)
Goat anti rabbit igg conjugated to fluorescent cy5 dye, by Abcam (1 mentions)
Hoxa1 antibodies, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Lsm 780 confocal microscope, by Zeiss (1 mentions)
Zen 2011, by Zeiss (1 mentions)
Lsm 780, by Zeiss (1 mentions)
Elyra ps 1, by Zeiss (1 mentions)
Plan apochromat 63 1.4 oil dic m27 objective, by Zeiss (1 mentions)
11 protocols using eclipse ti
1
Graphene Oxide Nanoparticles Influence on Neural Stem Cells
2
Encapsulating ECFCs in Stiff Hydrogels
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To increase matrix stiffness, mTG was added as a secondary cross-linker to laccase stock solutions to a concentration of 1.2 U/ml, as described previously (37 ) to achieve a final working concentration of 0.3 U/ml. ECFCs were encapsulated within these stiffer gels and monitored by both bright-field imaging (Olympus IX50) and time-lapse microscopy (Nikon Eclipse Ti or a Zeiss AXIO Observer Z.1) under standard cell culture conditions (37°C, 5% CO2). ImageJ (public domain) was used to quantify cell clusters.
3
Quantitative Microscopic Analysis of Bacteria
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Bacteria were imaged on agar pads by either conventional fluorescence microscopy (Nikon Eclipse E600, Nikon Eclipse Ti, or Zeiss Axioscope A1 with 100× objectives) or structured illumination microscopy (Nikon SIM-E/A1R with SR Apo TIRF 100× objective, UMass Amherst Light Microscopy Core).
To obtain the fluorescence intensity plots, the subcellular distribution of fluorescence was quantitated from images obtained by conventional fluorescence microscopy. The images were processed using Fiji and Oufti (Paintdakhi et al., 2016 (link); Schindelin et al., 2012 (link)) as described (García-Heredia et al., 2018 (link)). The signal was normalized to length and total fluorescence intensity of the cell. Cells were oriented such that the brighter pole is on the right hand of the graph. The intensity plots fromFigure 2A were made from 42Figure 3B, from 14To quantify the amount of fractionated cellular material in Figures 3 and 4 , images were processed in ImageJ, such that the cellular material corresponding to either the IMD or PM-CW fractions was measured. We then subtracted the signal from a constant-sized area of the gradient tubes that did not contain visible cellular material.
To obtain the fluorescence intensity plots, the subcellular distribution of fluorescence was quantitated from images obtained by conventional fluorescence microscopy. The images were processed using Fiji and Oufti (Paintdakhi et al., 2016 (link); Schindelin et al., 2012 (link)) as described (García-Heredia et al., 2018 (link)). The signal was normalized to length and total fluorescence intensity of the cell. Cells were oriented such that the brighter pole is on the right hand of the graph. The intensity plots from
4
Perfusion, Cryosectioning, and GAD67 Staining
5
Fluorescence Microscopy Imaging Protocol
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Sections under study were analyzed using epifluorescence (Nikon eclipse Ti) and confocal microscopy (Zeiss LSM 710). Brightness, contrast and gamma of images were adjusted using Irfanview software and figures arranged and lettered using Canvas software.
6
Immunohistochemical Analysis of Type I Collagen
7
Simultaneous Measurement of Intracellular and Mitochondrial ROS
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After treatment, cells were washed with cold PBS and then stained with two types of ROS probes [65 (link), 66 (link)]. DCFHDA (Molecular Probes, USA) was used to stain intracellular ROS while MitoSOX red mitochondrial superoxide indicator (Molecular Probes, USA) was used to stain mitochondrial ROS [67 (link)]. In brief, 5 μg/L DCFHDA and 2 μg/L MitoSOX were added to the medium and incubated for about 30 min in the dark. Cells were then washed three times with PBS and stained with DAPI, and images were taken on a Nikon Eclipse Ti epifluorescence microscope or a Zeiss LSM700 confocal microscope.
8
Collagen Localization in DPC Sheets
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DPC sheets were washed twice
with PBS and fixed in 10% formalin. Then, the cell sheets were processed
for standard paraffin embedding and sectioned at a 5 μm thickness.
Sections were stained with hematoxylin and eosin (H&E), or immunohistochemical
staining was performed using a primary antibody against type I collagen
(Abcam) and secondary antibody Alexa Fluor 488 anti-rabbit IgG (ThermoFisher).
DAPI staining was used to visualize nuclei. Images were captured using
a Nikon ECLIPSE Ti, ZEISS Scope.A1 AXIO or Nikon TE 2000 microscopes
and processed using the ImageJ software.
with PBS and fixed in 10% formalin. Then, the cell sheets were processed
for standard paraffin embedding and sectioned at a 5 μm thickness.
Sections were stained with hematoxylin and eosin (H&E), or immunohistochemical
staining was performed using a primary antibody against type I collagen
(Abcam) and secondary antibody Alexa Fluor 488 anti-rabbit IgG (ThermoFisher).
DAPI staining was used to visualize nuclei. Images were captured using
a Nikon ECLIPSE Ti, ZEISS Scope.A1 AXIO or Nikon TE 2000 microscopes
and processed using the ImageJ software.
9
Bacterial Imaging Methods Comparison
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Fixed bacteria were imaged either by conventional fluorescence microscopy (Nikon Eclipse E600, Nikon Eclipse Ti or Zeiss Axioscope A1 with 100x objectives) or by structured illumination microscopy (Nikon SIM-E/A1R with SR Apo TIRF 100x objective).
10
Chondrocyte HOXA1 Immunofluorescence Imaging
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Chondrocytes were grown in 20-mm glass-bottomed cell culture dishes (801001; Nest Biotechnology, Shanghai, China). The cells were fixed with 4% paraformaldehyde for 15 min, washed twice with PBS containing 0.05% Tween-20, permeabilized with 0.3% Triton X-100 for 5 min, and then blocked with 1% BSA for 30 min. The cells were then incubated with HOXA1 antibodies (1:200; Abcam) at 4°C overnight. After they were washed three times with PBS, the cells were incubated with goat anti-rabbit IgG conjugated to fluorescent Cy5 dye (1:100; Abcam) in PBS. DAPI (Life Technologies, Carlsbad, CA, USA) was used for nuclear staining. Immunofluorescence images were obtained using a Nikon Eclipse TI (Tokyo, Japan) and Zeiss LSM780 confocal microscope (Oberkochen, Germany) or Zeiss Colibri epifluorescence microscope and processed with ImageJ software (NIH, Bethesda, MD, USA).
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