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Lysosomes
Lysosomal dysfunction underlies a broad spectrum of pathologies—from inherited lysosomal storage diseases to neurodegeneration, cancer, and infectious disease. Researchers studying lysosomal biogenesis, autophagy flux, drug resistance mechanisms, or therapeutic targeting need reliable tools that report on lysosomal localization, pH dynamics, and membrane integrity without disrupting the processes under investigation.
AAT Bioquest's lysosome and autophagy probes address these needs across multiple experimental platforms. Our LysoBrite™ dyes exploit the lysosomal pH gradient for selective accumulation and pH-sensitive fluorescence enhancement, available in spectral variants from blue to NIR for multiplexing flexibility. CytoFix™ Red enables lysosomal tracking through fixation and downstream immunostaining workflows. For autophagy studies, our Autophagy Super Blue™ probe monitors autophagic activity in response to cellular stress, infection, or pharmacological intervention.
Fig. 1
HeLa cells were incubated in 1X HBSS buffer with 5% serum to induce starvation. Following starvation, cells were treated with Autophagy Green™ (Cat No. 23002) working solution for 20 minutes in a 37°C, 5% CO2 incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite™ Orange (Cat No. 22657).
LysoBrite™ Lysosomal Dyes
LysoBrite™ dyes are cell-permeable probes that selectively accumulate in lysosomes and acidic organelles via the lysosomal pH gradient. At nanomolar concentrations, pH-sensitive fluorescence enhancement upon lysosomal entry generates high-contrast signal suitable for fluorescence imaging, high-content screening, microplate fluorometry, or flow cytometry.
LysoBrite™ Orange, Red, Deep Red, and NIR variants offer excellent photostability and cellular retention for extended imaging and diverse applications including cell adhesion, chemotaxis, multidrug resistance, cell viability, apoptosis, and cytotoxicity studies.
Spectral Options
LysoBrite™ dyes span blue to near-infrared wavelengths for multiplexing with other fluorescent probes. For co-localization studies requiring fixation, use LysoBrite™ Red DND-99 (Cat No. 22647), which retains signal after aldehyde fixation. LysoBrite™ dyes are the core components of Cell Navigator® Lysosome Staining Kits (500 tests each).
Fig. 2
Images of HeLa cells stained with AAT Bioquest's LysoBrite™ Green (Cat No. 22643) and ThermoFisher's LysoTracker® Green DND-26 in a Costar black wall/clear bottom 96-well microplate. Samples were continuously illuminated for 120 seconds and the signals were compared before and after exposure using a Keyence fluorescence microscope.
Features
- Selective lysosomal accumulation via pH gradient
- pH-sensitive fluorescence for specific staining with minimal background
- Photostable signal for extended imaging
- Minimal cytotoxicity
- No-wash imaging protocol
- Compatible with proliferating and non-proliferating cells, suspension and adherent formats
CytoFix™ Red Fixable Lysosomal Stain
CytoFix™ Red maintains lysosomal localization through fixation—critical for experiments requiring immunostaining, long-term storage, or batch processing. Enhanced cellular retention preserves fluorescence signal for at least one week post-fixation, enabling longitudinal lysosomal tracking studies.
CytoFix™ Red multiplexes with GFP, other fluorescent conjugates, and organelle stains. Compatible with suspension and adherent cells across fluorescence microscopy, flow cytometry, and high-content platforms.
Fig. 3
The fluorescence images of HeLa cells stained with CytoFix™ LysoRed in a 96-well black-wall clear-bottom plate. Image was acquired before (Left) and after (Right) fixation with 4% formaldehyde solution for 20 minutes at RT. The cells were imaged using fluorescence microscope with a Cy3/TRITC filter.
Features
- High lysosomal staining efficiency
- Signal retention ≥1 week after fixation
- Minimal hands-on time
Autophagy Probes
Autophagy is a lysosomal degradation pathway that sequesters cytoplasmic cargo into double-membrane autophagosomes, which subsequently fuse with lysosomes for content digestion and recycling. As a protective mechanism against cellular stress, infection, and metabolic dysfunction, autophagy is a key target in cancer, neurodegeneration, and infectious disease research.
Cell Meter™ Autophagy Fluorescence Imaging Kits use Autophagy Super Blue™ probe to selectively target autophagosomes and monitor autophagy activity in live cells responding to stress, microbial infection, or pharmacological modulation.
Fig. 4
Autophagy Green™ labeled vesicles are induced by starvation in HeLa cells. HeLa cells were incubated in a regular DMEM medium (Control) or in 1X HBSS buffer with 5% serum (Autophagy Treatment) for 16 hours. Both control cells and starved cells were incubated with Autophagy Green™ working solution for 20 minutes in a 37°C, 5% CO2 incubator, and then washed 3 times with wash buffer. Cells were imaged immediately under a fluorescence microscope with a FITC channel (green). Cell nuclei were stained with Hoechst 33342 (Cat No. 17530).
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Document: 01.0085.211015r1
Last updated Wed Dec 10 2025