Sections were qualitatively analyzed by using both bright- and darkfield microscopes (Leica DMR; Perkin Elmer UltraView Vox spinning-disk confocal on a Zeiss Axiovert). The relative density and intensity of Celf6 expression in WT, Celf6-YFP and Celf6−/− tissues were determined using Leica DMR microscrope. Brightfield images were captured on a Leica DFC310FX camera mounted to a Leica DM400B microscope using Surveyor software version V7.0.0.6 MT (Objective Imaging, Cambridge, UK) and darkfield images captured on a Perkin Elmer UltraView Vox spinning-disk confocal on a Zeiss Axiovert microscope. Digital raw images were optimized for evenness of illumination and background by using ImageJ (Wayne Rasband, NIH, USA). Anatomical structures were identified according to an adult mouse brain atlas (Franklin and Paxinos). Overall expression was scored by a rubric using both density of cells labeled (+, scattered; ++, light; +++, moderate, or ++++, high) and intensity of signal (+, weak; ++, moderate; or +++, strong). The overall expression score for each region was calculated by multiplying density level by signal intensity. Thus, the expression scores indicate the following: 1 – 2, light expression; 3 – 6, moderate expression; 8 – 9, high expression; and 12, very high expression.
Ultraview vox
Manufactured by Zeiss
Sourced in United Kingdom
The UltraVIEW VoX is a confocal imaging system designed for live-cell imaging and high-resolution analysis. It provides a versatile platform for advanced fluorescence microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
Dfc310 fx, by Leica (1 mentions)
Dm400b, by Leica (1 mentions)
Observer z1, by Zeiss (1 mentions)
Comet assay kit, by Abcam (1 mentions)
Csu x1, by Yokogawa (1 mentions)
Imageem c9100 13 emccd camera, by Hamamatsu Photonics (1 mentions)
Axio observer microscope, by Yokogawa (1 mentions)
Mtt cell proliferation and cytotoxicity assay kit, by Solarbio (1 mentions)
Fm4 64, by Thermo Fisher Scientific (1 mentions)
Uv 2600, by Shimadzu (1 mentions)
Dicoumarol, by Maclin Biochemical Technology (1 mentions)
Mitotracker red, by Beyotime (1 mentions)
4 protocols using ultraview vox
1
Quantifying Celf6 Expression in Mouse Brain
2
DNA Damage Evaluation in MCF-7 Cells
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MCF-7 cells were seeded
at a density of
1.0 × 106 cells into a 25 cm2 flask, followed
by 24 h of incubation in a humidified atmosphere (5% CO2/95% air) at 37 °C. Freshly prepared stock solutions ofGW7604-Pent-PtCl 3 and Cisplatin in
DMF were used and diluted with the medium to reach a final complex
concentration corresponding to their IC50 values of 20
μM and 15 μM, respectively. Cells were incubated with
the compounds for 48 h. The assay was performed according to the manufacturer’s
protocol (Comet assay kit, Abcam, Cambridge, UK). Briefly, after lysis
and application to the microscopy slides, electrophoresis was run
in an alkaline buffer for 20 min at 2 V/cm. Subsequently, slides were
washed with pure deionized water and EtOH/water [7/3 (v/v)], and stained with Vista Green DNA Dye. Visualization
was carried out by real-time live confocal microscopy using an inverted
microscope (Zeiss Observer Z1) in arrangement with a spinning disc
confocal system (UltraVIEW VoX).
at a density of
1.0 × 106 cells into a 25 cm2 flask, followed
by 24 h of incubation in a humidified atmosphere (5% CO2/95% air) at 37 °C. Freshly prepared stock solutions of
DMF were used and diluted with the medium to reach a final complex
concentration corresponding to their IC50 values of 20
μM and 15 μM, respectively. Cells were incubated with
the compounds for 48 h. The assay was performed according to the manufacturer’s
protocol (Comet assay kit, Abcam, Cambridge, UK). Briefly, after lysis
and application to the microscopy slides, electrophoresis was run
in an alkaline buffer for 20 min at 2 V/cm. Subsequently, slides were
washed with pure deionized water and EtOH/water [7/3 (v/v)], and stained with Vista Green DNA Dye. Visualization
was carried out by real-time live confocal microscopy using an inverted
microscope (Zeiss Observer Z1) in arrangement with a spinning disc
confocal system (UltraVIEW VoX).
3
FRET Imaging of Contractile Ring Proteins
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FRET imaging (Figure 5 ) was performed on a Perkin Elmer Ultraview Vox spinning disk system equipped with a Zeiss Axio Observer microscope, 488 and 561 nm solid state lasers with a PhotoKinesis bleaching module, a Yokogawa CSU-X1 spinning disk, a 63X C-Apochromat objective, and a Hamamatsu ImageEM C9100-13 EMCCD camera. An acceptor photobleaching method was employed for FRET imaging in live cells. The mCherry-Cdc15 or Cdc15-mCherry fluorophores were bleached throughout the cells with 100% 561 nm laser power for 20 cycles. 10 frames of single Z slices in the donor GFP channel were acquired pre- and post-bleach. FRET percentages were calculated by first correcting for background and photobleaching over the 10 pre- and post-bleach frames, and subsequently calculating the percentage increase in GFP donor fluorescence at contractile ring ROIs of a consistent size. Statistical tests in Figure 5A–B and Figure 5—figure supplement 1B and S5B are ANOVA tests versus Rlc1-GFP a negative control which does not FRET with either mCherry-Cdc15 or Cdc15-mCherry, with uncorrected p values reported in Figure 5—figure supplement 1A–B . Statistical tests in Figure 5—figure supplement 1C were Fisher’s exact tests between the mCherry-Cdc15 and Cdc15-mCherry conditions.
4
Comprehensive Characterization of Biomaterials
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All common starting chemical reagents and solvents were used as purchased without further purification. NTRs, NADH, MTZ, and Gal were purchased from Sigma-Aldrich. Dicoumarol was purchased from Macklin. OXA, DOX, PTX, IRT, and 5-Fu were purchased from Aladdin Biotech Co. Ltd. 3-MA was purchased from Selleck Co. MitoTracker Red was purchased from Beyotime Biotechnology. MTT Cell Proliferation and Cytotoxicity Assay Kit was purchased from Beijing Solarbio Science & Technology. FM 4-64 and PI were purchased from Thermo Fisher Scientific for bacterial live/dead staining.
Nuclear magnetic resonance spectra were recorded on a Bruker 400-MHz Fourier transform spectrometer. TEM images were collected on a Tecnai G2 20 S-TWIN and HT7700 transmission electron microscope. SEM images were captured on a Merlin scanning electron microscope. MALDI-TOF mass spectra were recorded on a Bruker AutoFlex Max mass spectrometer. The fluorescence spectra were recorded on F98, and UV-vis spectra were obtained on Shimadzu UV-2600. The CLSM images were captured with a confocal microscope (Zeiss 710 and UltraVIEW VoX).
Nuclear magnetic resonance spectra were recorded on a Bruker 400-MHz Fourier transform spectrometer. TEM images were collected on a Tecnai G2 20 S-TWIN and HT7700 transmission electron microscope. SEM images were captured on a Merlin scanning electron microscope. MALDI-TOF mass spectra were recorded on a Bruker AutoFlex Max mass spectrometer. The fluorescence spectra were recorded on F98, and UV-vis spectra were obtained on Shimadzu UV-2600. The CLSM images were captured with a confocal microscope (Zeiss 710 and UltraVIEW VoX).
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