Lysosomes are membrane-bound organelles that are involved in several cellular processes such as biomolecule degradation, apoptosis, cell signaling, metabolism, and plasma membrane repair. Lysosome markers are effective tools for identifying and tracking lysosomes. While LysoTracker markers target acid contents and localize within lysosomes, CellLight fluorescent fusion proteins target to the lysosomal membrane for locating and tracking of lysosomes.
Lysosomes are membrane-bound organelles that contain enzymes which degrade biomolecules such as carbohydrates, lipids, nucleic acids, and peptides [1–3]. Intracellular molecules are digested through a process called autophagy while extracellular material that are taken up by endocytosis are digested through a process called phagocytosis [1, 2]. Lysosomes are also involved in the digestion of viral particles or bacteria through phagocytosis. Lysosomes have been shown to be involved in other cellular processes including intracellular transport of internal and external material, signaling to regulate proliferation and growth, and cellular metabolism .
|Target||Targets acidic organelles|
|Readout||Localization of lysosomes by fluorescence imaging|
|Common filter set||DAPI||FITC||TRITC||Cy5|
|Labels||LysoTracker Blue||LysoTracker Green||LysoTracker Red||LysoTracker Deep Red|
|Format||20 x 50 uL||20 x 50 uL||20 x 50 uL||5 x 50 uL|
LysoTrackers are cell-permeable lysosome markers that target and track acidic organelles in live cells. LysoTracker markers are comprised of a fluorophore bound to weakly basic amines. Though the mechanism of LysoTracker retention is relatively unknown, it is likely due to protonation with retention in the organelles’ membrane. These lysosomal markers accumulate in organelles that have a low pH and are only partially protonated at neutral pH. LysoTracker lysosomal markers are available in a range of colors to allow for multiplexing with other fluorescent markers (Figures 1 and 2).
CellLight fluorescent fusion proteins are lysosome membrane constructs that label lysosomes in live cells to follow the dynamics of intracellular behavior. CellLight fluorescent proteins use the Lamp1 (lysosomal associated membrane protein 1) construct fused to emGFP (Figure 3) or TagRFP (Figure 4). CellLight lysosome membrane markers are available in green or red fluorescent fusion proteins and can be multiplexed with other fluorescent dyes and proteins. Additionally, fluorescent staining is retained after fixation and permeabilization.
Lysosome membrane markers also serve as tools for tracking fusion with the autophagosome prior to degradation of the autolysosome.
Figure 4. Live cell imaging with CellLight fluorescent fusion proteins. Cascade Biologics human aortic smooth muscle cells (HASMC) were transduced with CellLight Lysosomes-RFP, CellLight MAP4-GFP, and Hoechst 33342. Imaging was performed on live cells using a DeltaVision Core microscope and standard DAPI/FITC/TRITC filter sets.
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