Nuc-Blue live cell nuclear stain (Invitrogen, UK) was applied to the constrained and unconstrained tissue blocks in order to visualise cell nuclei. A phase contrast background was used to locate cell nuclei within the surrounding matrix. Tissue blocks were observed under 20× objective in a wide-field microscope with incubator for maintaining the standard culture conditions. ‘Volocity’ 3D image analysis software was used to record time-lapse video of all disc samples at 6, 12, and 36 h.
Nucblue live cell nuclear stain
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
NucBlue live cell nuclear stain is a fluorescent dye used to label the nuclei of living cells. It binds to DNA and emits blue fluorescence when excited by appropriate light.
Automatically generated - may contain errors
Lab products found in correlation
Evos fluorescence microscope, by Thermo Fisher Scientific (2 mentions)
Zen lite 2012 sp2, by Zeiss (2 mentions)
Live dead violet dye lvd, by Thermo Fisher Scientific (2 mentions)
Axio observer z1, by Zeiss (2 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Cytochalasin d, by Cayman Chemical (1 mentions)
Bafilomycin a1, by Abcam (1 mentions)
Celltracker deep red, by Thermo Fisher Scientific (1 mentions)
Cilengitide, by Bio-Techne (1 mentions)
Live cell imaging solution, by Thermo Fisher Scientific (1 mentions)
Jasplakinolide, by Santa Cruz Biotechnology (1 mentions)
Live dead fixable aqua stain, by Thermo Fisher Scientific (1 mentions)
Psb0739, by Bio-Techne (1 mentions)
Mrs2179, by Bio-Techne (1 mentions)
Adenosine diphosphate (adp), by Merck Group (1 mentions)
Mrs2211, by Bio-Techne (1 mentions)
Fluo 4 am, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
5 protocols using nucblue live cell nuclear stain
1
Nuclear Staining and Imaging of Tissue Samples
2
Multi-parameter Live Cell Imaging
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Live Cell Imaging solution, Fluo4-AM, CellTracker Deep Red, pHrodo iFL Red STP Ester, NucBlue live cell nuclear stain, NucGreen dead cell nuclear stain, and LIVE/DEAD fixable aqua stain were obtained from Invitrogen. DAPI was from Sigma, and paraformaldehyde (4% in PBS) was obtained from Alfa Aesar. Human annexin V protein was from BD Biosciences, human C5a protein from Peprotech, human TGFβ1 from Miltenyi Biotec, and recombinant human IFNγ and recombinant truncated human vitronectin from Gibco. Cytochalasin D was from Cayman Chemicals, Bafilomycin A1 from Abcam, and Jasplakinolide from Santa Cruz. PSB0739, MRS2179, MRS2211, and cilengitide were all obtained from Tocris. ADP, BSA, and crystal violet were from Sigma.
3
Imaging YFP-Gal8 in MDA-MB-231 Cells
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MDA-MB-231 cells expressing YFP-Gal8 were generated as previously published68 (link) and plated in a black 96-well plate 24 h prior to treatment with media only, lipofectamine RNAiMax with 50 nM “zipper” siRNA against luciferase, or 1 μM of siLuc < (EG18L)2 in Opti-MEM with NucBlue live cell nuclear stain (Invitrogen). Cells were imaged at 18 h after initiation of treatment using a Nikon Ti confocal microscope.
4
Fluorescence Imaging of C. neoformans Strains
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C. neoformans WT and mutant strains were evaluated for fluorescence emission and the proper cellular compartment localization of histone H3 or Pab, tagged with GFP or mCherry, using a Zeiss Axio Observer Z1 inverted phase-contrast fluorescence microscope (Carl Zeiss) equipped with a 63× 1.4-numerical-aperture (NA) oil immersion objective, the high efficiency (HE) filters 38 HE (green) and 63 HE (red), and differential interference contrast (DIC). Images were captured using ZEN lite 2012 SP2 software (Carl Zeiss). Viable but nonculturable yeast cells were imaged using an Evos fluorescence microscope (Thermo Fisher) with 357/44-nm (4′,6-diamidino-2-phenylindole [DAPI]) and 470/22-nm (GFP) LED lights coupled to 447/60- and 510/42-nm emission filters, respectively.
For the visualization of nuclei, cells were stained with NucBlue live-cell nuclear stain (Thermo Fisher) for 5 min. For viability analysis, cells were incubated with LIVE/DEAD violet dye (LVD) (excitation/emission at 405/450 nm) (Thermo Fisher) for 20 min at room temperature with protection from light. As a control, dead cells were used after heat inactivation for 1 h at 70°C (heat killed [HK]). Cells were washed twice prior to analysis.
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For the visualization of nuclei, cells were stained with NucBlue live-cell nuclear stain (Thermo Fisher) for 5 min. For viability analysis, cells were incubated with LIVE/DEAD violet dye (LVD) (excitation/emission at 405/450 nm) (Thermo Fisher) for 20 min at room temperature with protection from light. As a control, dead cells were used after heat inactivation for 1 h at 70°C (heat killed [HK]). Cells were washed twice prior to analysis.
5
Fluorescence Imaging of C. neoformans Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
C. neoformans WT and mutant strains were evaluated for fluorescence emission and the proper cellular compartment localization of histone H3 or Pab, tagged with GFP or mCherry, using a Zeiss Axio Observer Z1 inverted phase-contrast fluorescence microscope (Carl Zeiss) equipped with a 63× 1.4-numerical-aperture (NA) oil immersion objective, the high efficiency (HE) filters 38 HE (green) and 63 HE (red), and differential interference contrast (DIC). Images were captured using ZEN lite 2012 SP2 software (Carl Zeiss). Viable but nonculturable yeast cells were imaged using an Evos fluorescence microscope (Thermo Fisher) with 357/44-nm (4′,6-diamidino-2-phenylindole [DAPI]) and 470/22-nm (GFP) LED lights coupled to 447/60- and 510/42-nm emission filters, respectively.
For the visualization of nuclei, cells were stained with NucBlue live-cell nuclear stain (Thermo Fisher) for 5 min. For viability analysis, cells were incubated with LIVE/DEAD violet dye (LVD) (excitation/emission at 405/450 nm) (Thermo Fisher) for 20 min at room temperature with protection from light. As a control, dead cells were used after heat inactivation for 1 h at 70°C (heat killed [HK]). Cells were washed twice prior to analysis.
+ Expand
For the visualization of nuclei, cells were stained with NucBlue live-cell nuclear stain (Thermo Fisher) for 5 min. For viability analysis, cells were incubated with LIVE/DEAD violet dye (LVD) (excitation/emission at 405/450 nm) (Thermo Fisher) for 20 min at room temperature with protection from light. As a control, dead cells were used after heat inactivation for 1 h at 70°C (heat killed [HK]). Cells were washed twice prior to analysis.
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