Lysoview 633

Manufactured by Biotium

Lysoview 633 is a fluorescent dye used for labeling and visualizing lysosomes in live cells. It is a bright, photostable dye that selectively stains acidic organelles, allowing for the observation of lysosomal dynamics and distribution within the cell.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Monensin, by Bio-Techne (1 mentions) Propidium iodide, by ImmunoChemistry Technologies (1 mentions) Takinib, by MedChemExpress (1 mentions) Filipin 3, by Merck Group (1 mentions) Ultrapure lps, by InvivoGen (1 mentions) Dq green bsa, by Thermo Fisher Scientific (1 mentions) Mcc950, by InvivoGen (1 mentions) Cellbrite green, by Biotium (1 mentions) Rpmi medium, by Thermo Fisher Scientific (1 mentions) Zeocin, by InvivoGen (1 mentions) M csf, by R&D Systems (1 mentions) Lovastatin, by Cayman Chemical (1 mentions) Digitonin, by Merck Group (1 mentions) Imipramine hydrochloride imp, by Merck Group (1 mentions) Hydroxychloroquine sulfate hcq, by Merck Group (1 mentions) Filipin, by Merck Group (1 mentions) Methyl β cyclodextrin mβcd, by Merck Group (1 mentions) Fluorodish, by Thermo Fisher Scientific (1 mentions)

5 protocols using lysoview 633

Inflammasome Activation Assay Protocol

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All reagents and solvents
were purchased from
commercial suppliers (Sigma-Aldrich, Alfa Aesar, Fluka, TCI America).
Ultrapure LPS, anti-IL-1β antibody, isoliquiritigenin, and MCC950
were purchased from Invivogen. All cell media, Hoechst 33342, DQ Green
BSA, and Alexa Flour 488-NHS were bought from Thermo Fischer Scientific.
Lysoview 633 was obtained from Biotium. APG-4 was purchased from TEFLabs,
monensin from Tocris, and filipin-III from Sigma-Aldrich. Propidium
iodide, FAM-YVAD-FMK kit (FAM FLICA Caspase-1), and cathepsin B MM-(RR)₂ kit (Magic Red cathepsin B) were bought from Immunochemistry
Technologies. Ca-074-Me was obtained from Calbiochem. E 64d and (5Z)-7-Oxozeaenol
were from Tocris. Takinib was from Medchemexpress. Rabbit polyclonal
anti-TMS1 and donkey antirabbit IgG Alexa Fluor 647 antibodies were
purchased from Abcam. All peptide synthesis reagents were purchased
from Novabiochem. Fmoc-N-amido-dPEG₆-acid linker was purchased
from Quanta Biodesign, and azidohexanoic acid was purchased from Click
Chemistry Tools.

Manna S., Howitz W.J., Oldenhuis N.J., Eldredge A.C., Shen J., Nihesh F.N., Lodoen M.B., Guan Z, & Esser-Kahn A.P. (2018). Immunomodulation of the NLRP3 Inflammasome through Structure-Based Activator Design and Functional Regulation via Lysosomal Rupture. ACS Central Science, 4(8), 982-995.

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Glucose-Induced Lysosome Imaging in HepG2 Cells

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HepG2 cells were plated in a 6- or 12-well plate after being strained through a 70 μm cell strainer and grown into a monolayer for 24 h. On the day of the experiment, cells were serum-starved for 3-6 h and then pretreated with 30 μM metformin or a PBS vehicle for 35 min before supplementing 5- or 25 mM D-glucose. 30 min following D-glucose supplementation LysoView™ 633 at the final concentration of 1X and CellBrite® Green (Biotium) diluted 1:200 was added to each well and incubated for 30 min at 37° C then imaged using an EVOS fluorescent imaging system with the Cy5 and GFP filters.

Senfeld J., Peng Q., Shi Y., Qian S, & Shen J. (2023). A purinergic mechanism underlying metformin regulation of hyperglycemia. iScience, 26(6), 106898.

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Macrophage Differentiation and Lysosome Monitoring

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Phorbol-Myristate-Acetate (PMA, provider), RPMI medium (Gibco), Fetal bovine serum (thermofisher), human off-the-clot serum (Geminibio), human recombinant macrophage-colony stimulating factor (M-CSF, R&D Systems), Lysoview-633 (biotium), OADC supplement (R&D Systems), Zeocin (invivogen).

Augenstreich J., Poddar A., Belew A.T., El-Sayed N.M, & Briken V. (2024). da_Tracker: Automated workflow for high throughput single cell and single phagosome tracking in infected cells. bioRxiv.

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Cellular Trafficking Pathway Modulators

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Imipramine hydrochloride (IMP; Sigma, 10899–5G), hydroxychloroquine sulfate (HCQ; Sigma, H0915–5MG), fluvoxamine maleate (FLV; Cayman, 15617), fluoxetine hydrochloride (FLX; Enzo, BML-NS140-0050), bafilomycin (BAF; EnzoLife Sciences, BML-CM110-0100), U18666A (EMD Millipore, 662015–10MG), methyl-β-cyclodextrin (MβCD; Sigma, C4555), filipin (Sigma, F9765), digitonin (Sigma, 300410), Lysoview 633 (Biotium, 70058), lovastatin (Cayman, 10010338).

Kendall R.L, & Holian A. (2023). Cholesterol-dependent molecular mechanisms contribute to cationic amphiphilic drugs’ prevention of silica-induced inflammation. European journal of cell biology, 102(2), 151310.

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Cellular Internalization and Lysosomal Colocalization

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Fluorescence images were taken on a Lecia SP5 II Scanning Confocal Microscope. For internalization experiments, cells were plated at a density of 50,000 cells/dish on a FluoroDish (Fisher, 35100) in 500 μL of 2% serum media. Once 60–70% confluent, cells were treated with 20nM CoGliF carried by 3:1 or 1:4 CoGliF/carrier weight ratio of PEI or GOPEI, respectively. For lysosomal staining, 1 μL of 1000x LysoView 633 (Biotium, 70058) was added to the media for 1 hr. Media was swapped after 4x washing with DPBS, and DAPI stain was added 10 minutes before imaging. Percent colocalization with lysosomes was determined using ImageJ software analysis. Background signals were excluded from the threshold in both images. A selection was created using the signal in the CoGliF channel and was superimposed onto the LysoView channel. The percent colocalization was calculated by the percentage of CoGliF pixels that overlap with LysoView 633 pixels. Data were plotted in GraphPad Prism and represent mean ± one standard deviation. T-test comparisons were used to determine the statistical significance of the resulting changes in colocalization from 0 hours post-CoGliF removal to 24 hours post removal.

Dukes M.W., Bajema E.A., Whittemore T.J., Holmgren R.A, & Meade T.J. (2022). Delivery of Targeted Co(III)-DNA Inhibitors of Gli Proteins to Disrupt Hedgehog Signaling. Bioconjugate chemistry, 33(4), 643-653.

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