Cell Navigator® Lysosome Staining Kit *Red Fluorescence*
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Excitation (nm) | 576 |
Emission (nm) | 596 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Excitation (nm) 576 | Emission (nm) 596 |
Platform
Fluorescence microscope
Excitation | TRITC filter |
Emission | TRITC filter |
Recommended plate | Black wall/clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add LysoBrite™ Red working solution
- Incubate at 37°C for 30 minutes
- Wash the cells
- Analyze the cells under fluorescence microscope at Ex/Em = 575/600 nm (TRITC filter set)
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 20 µL of 500X LysoBrite™ Red (Component A) to 10 mL of Live Cell Staining Buffer (Component B) to make LysoBrite™ Red working solution. Protect from light. Note: 20 µL of 500X LysoBrite™ Red (Component A) is enough for one 96-well plate. The optimal concentration of the fluorescent lysosome indicator varies depending on the specific application. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
For adherent cells:
- Grow cells either in a 96-well black wall/clear bottom plate (100 µL/well/96-well plate) or on cover-slips inside a petri dish filled with the appropriate culture medium.
- When cells reach the desired confluence, add equal volume of LysoBrite™ Red working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes.
- Wash the cells twice with pre-warmed (37°C) Hanks and 20 mM Hepes buffer (HBSS) or buffer of your choice, fill the cell wells with HBSS or growth medium.
- Observe the cells using a fluorescence microscope with TRITC filter set (Ex/Em = 575/600 nm). Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
For suspension cells:
- Add equal volume of LysoBrite™ Red working solution into the cells.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes.
- Wash the cells twice with pre-warmed (37°C) Hanks and 20 mM Hepes buffer (HBSS) or buffer of your choice, fill the cell wells with HBSS or growth medium.
- Observe the cells using a fluorescence microscope with TRITC filter set (Ex/Em = 575/600 nm). Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained. Suspension cells may be attached to cover-slips that have been treated with BD Cell-Tak® (BD Biosciences) and stained as adherent cells.
Product Family
Images
![Images of HeLa cells stained with A: Cell Navigator® Lysosome Staining Kit (Cat# 22658), B: LysoTracker® Red DND-99 (from Invitrogen) in a Costar black wall/clear bottom 96-well plate. The signals were compared at 0 and 120 seconds exposure time by using an Olympus fluorescence microscope.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-lysosome-staining-kit-red-fluorescence_FSspV.jpg&w=3840&q=75)
![Image of Hela cells stained with Cell Navigator® Lysosome Staining Kit in a Costar black wall-clear bottom 96-well plate.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-lysosome-staining-kit-red-fluorescence_gbra0.jpg&w=3840&q=75)
![Live PDAC microtumours feed on dead-cell debris. (a,b) Time-lapse images of live and dead cell assays. A live PCI-55 microtumour showed that an early apoptotic cell underwent late apoptosis on the surface of microtumours, and was then taken into their bodies (a). PCI-55 microtumours that took up Annexin V<sup>+</sup>/EthD-1<sup>+</sup> cells massively accumulated Annexin V on their surfaces, whereas EthD-1 was dispersed throughout their bodies (b). (c–i) Feeding dead-cell debris to anchorage-dependent PCI-55 microtumours. To maintain PCI-55 microtumours anchored to the micro/nanoplate, we added their UV-induced dead-cell debris. Schematic of the protocol (c). Time-lapse images (d). Time after adding dead-cell debris shown as hh:mm. Grown microtumours devoured dead-cell debris aggressively (e). Tumour sizes in PCI-55 microtumours at 48 h after dead-cell feeding (f–h), CFSE-labelled PCI-55 microtumours were fed UV-induced dead PCI-55 cells into which Edu (red fluorescence) had been incorporated. 3D images of CFSE-labelled PCI-55 microtumours with added Edu<sup>+</sup> dead-cell debris (g). Confocal images of CFSE-labelled PCI-55 microtumours with added Edu<sup>+</sup> dead-cell debris. Cross sections of CFSE-labelled PCI-55 microtumours taking up debris-derived Edu (h). Fluorescence for indicated markers of PCI-55 microtumours taking up debris-derived Edu (i). Zoomed-in image shows that Edu<sup>+</sup> dead-cell debris was endocytosed from the surface of the PDAC microtumour and then incorporated into lysosomes in the inner cells forming the microtumours. Source: <strong>Visualising the dynamics of live pancreatic microtumours self-organised through cell-in-cell invasion</strong> by Miyatake et al., <em>Scientific Reports</em>, Sept. 2018.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-lysosome-staining-kit-red-fluorescence_gyq2i.jpg&w=3840&q=75)
![Lysosome localization and motility is altered for starvation-induced Hela cells. A: Healthy untreated Hela cells. Lysosomes (Red) were dispersed widely throughout the cytosol in cells. B: Starved Hela Cells. The cells were starved for 24 hours (no serum), and lysosomes were aggregated in the perinuclear region. Nuclei were stained with Hoechst 33342.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-red-fluorescence%2Ffigure-for-cell-navigator-lysosome-staining-kit-red-fluorescence_drqK6.jpg&w=3840&q=75)
Citations
Authors: Ben-Akiva, Elana and Karlsson, Johan and Hemmati, Shayan and Yu, Hongzhe and Tzeng, Stephany Y and Pardoll, Drew M and Green, Jordan J
Journal: Proceedings of the National Academy of Sciences (2023): e2301606120
Authors: Wu, Xuanjin and Li, Yang and Chen, Xiguang and Zhou, Zhongzheng and Pang, Jianhui and Luo, Xin and Kong, Ming
Journal: Journal of Materials Chemistry B (2019): 4854--4866
Authors: Martin, Victor and Ribeiro, Isabel AC and Alves, Marta M and Gon{\c{c}}alves, L{\'\i}dia and Almeida, Ant{\'o}nio J and Grenho, Liliana and Fernandes, Maria H and Santos, Catarina F and Gomes, Pedro S and Bettencourt, Ana F
Journal: International journal of pharmaceutics (2019): 118821
Authors: Miyatake, Yukiko and Kuribayashi-Shigetomi, Kaori and Ohta, Yusuke and Ikeshita, Shunji and Subagyo, Agus and Sueoka, Kazuhisa and Kakugo, Akira and Amano, Maho and Takahashi, Toshiyuki and Okajima, Takaharu and others, undefined
Journal: Scientific reports (2018): 14054
Authors: Bai, Xiaoyu and Kong, Ming and Wu, Xuanjin and Feng, Chao and Park, Hyunjin and Chen, Xiguang
Journal: Journal of Materials Chemistry B (2018): 5910--5921
Authors: Lechevallier, S{\'e}verine and Mauricot, Robert and Gros-Dagnac, H{\'e}l{\`e}ne and Chevreux, Sylviane and Lemercier, Gilles and Phonesouk, Erick and Golzio, Muriel and Verelst, Marc
Journal: ChemPlusChem (2017): 770--777
Authors: Paris, Juan L and de la Torre, Paz and Cabanas, M Victoria and Manzano, Miguel and Grau, Montserrat and Flores, Ana I and Vallet-Reg{\'\i}, Mar{\'\i}a
Journal: Nanoscale (2017): 5528--5537
Authors: Liao, Wei-Chih and Huang, Mei-Zi and Wang, Michelle Lily and Lin, Chun-Jung and Lu, Tzu-Li and Lo, Horng-Ren and Pan, Yi-Jiun and Sun, Yu-Chen and Kao, Min-Chuan and Lim, Hui-Jing and others,
Journal: Frontiers in cellular and infection microbiology (2017): 203
Authors: Lechevallier, Séverine and Mauricot, Robert and Gros-Dagnac, Hélène and Chevreux, Sylviane and Lemercier, Gilles and Phonesouk, Erick and Golzio, Muriel and Verelst, Marc
Journal: ChemPlusChem (2017)
Authors: Wilson, David R and Routkevitch, Denis and Rui, Yuan and Mosenia, Arman and Wahlin, Karl J and Quinones-Hinojosa, Alfredo and Zack, Donald J and Green, Jordan J
Journal: Molecular Therapy (2017)
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