Limits...
Lysosomal targeting with stable and sensitive fluorescent probes (Superior LysoProbes): applications for lysosome labeling and tracking during apoptosis.

Chen X, Bi Y, Wang T, Li P, Yan X, Hou S, Bammert CE, Ju J, Gibson KM, Pavan WJ, Bi L - Sci Rep (2015)

Bottom Line: The use of Superior LysoProbes facilitates the direct visualization of the lysosomal response to lobaplatin elicited in human chloangiocarcinoma (CCA) RBE cells, using confocal laser scanning microscopy.Additionally, we have characterized the role of lysosomes in autophagy, the correlation between lysosome function and microtubule strength, and the alteration of lysosomal morphology during apoptosis.Our findings indicate that Superior LysoProbes offer numerous advantages over previous reagents to examine the intracellular activities of lysosomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Michigan Technological University, Houghton, MI 49931.

ABSTRACT
Intracellular pH plays an important role in the response to cancer invasion. We have designed and synthesized a series of new fluorescent probes (Superior LysoProbes) with the capacity to label acidic organelles and monitor lysosomal pH. Unlike commercially available fluorescent dyes, Superior LysoProbes are lysosome-specific and are highly stable. The use of Superior LysoProbes facilitates the direct visualization of the lysosomal response to lobaplatin elicited in human chloangiocarcinoma (CCA) RBE cells, using confocal laser scanning microscopy. Additionally, we have characterized the role of lysosomes in autophagy, the correlation between lysosome function and microtubule strength, and the alteration of lysosomal morphology during apoptosis. Our findings indicate that Superior LysoProbes offer numerous advantages over previous reagents to examine the intracellular activities of lysosomes.

No MeSH data available.


Related in: MedlinePlus

Intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to MitoTracker (80 nM) and LysoTracker Green (2 μM).HeLa cells were imaged on an inverted laser scanning fluorescent microscope (Olympus) using a 60 × oil immersion objective lens.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4355733&req=5

f2: Intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to MitoTracker (80 nM) and LysoTracker Green (2 μM).HeLa cells were imaged on an inverted laser scanning fluorescent microscope (Olympus) using a 60 × oil immersion objective lens.

Mentions: To examine the intracellular localization of the synthesized probes, double staining with MitoTracker in HeLa cells was performed. Separate labeling patterns, and a lack of co-localization of the probe with mitochondria, were observed after 30 min incubation (Fig. 2 and Fig. S2–S4). To further explore labeling patterns, HeLa cells were stained with LysoTracker Green (a commercial lysosome selective stain), which revealed colocalization of Superior LysoProbe IV with lysosomes (Fig. 2). Hoechst 33342 was employed for nucleus staining, in order to confirm cell viability throughout the experiment. To estimate the degree of colocalization of Superior LysoProbes and LysoTracker, a quantitative analysis of confocal images was performed. The Pearson's correlation coefficient (POC) and the Mander's overlap coefficient (MOC) were used to quantify the degree of co-localization between fluorophores. The Pearson's correlation coefficients for Superior LysoProbe I–IV and LysoTracker were high. Additionally, the overlap coefficients of the two fluorescence patterns were also relatively high (Table S1).


Lysosomal targeting with stable and sensitive fluorescent probes (Superior LysoProbes): applications for lysosome labeling and tracking during apoptosis.

Chen X, Bi Y, Wang T, Li P, Yan X, Hou S, Bammert CE, Ju J, Gibson KM, Pavan WJ, Bi L - Sci Rep (2015)

Intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to MitoTracker (80 nM) and LysoTracker Green (2 μM).HeLa cells were imaged on an inverted laser scanning fluorescent microscope (Olympus) using a 60 × oil immersion objective lens.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4355733&req=5

f2: Intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to MitoTracker (80 nM) and LysoTracker Green (2 μM).HeLa cells were imaged on an inverted laser scanning fluorescent microscope (Olympus) using a 60 × oil immersion objective lens.
Mentions: To examine the intracellular localization of the synthesized probes, double staining with MitoTracker in HeLa cells was performed. Separate labeling patterns, and a lack of co-localization of the probe with mitochondria, were observed after 30 min incubation (Fig. 2 and Fig. S2–S4). To further explore labeling patterns, HeLa cells were stained with LysoTracker Green (a commercial lysosome selective stain), which revealed colocalization of Superior LysoProbe IV with lysosomes (Fig. 2). Hoechst 33342 was employed for nucleus staining, in order to confirm cell viability throughout the experiment. To estimate the degree of colocalization of Superior LysoProbes and LysoTracker, a quantitative analysis of confocal images was performed. The Pearson's correlation coefficient (POC) and the Mander's overlap coefficient (MOC) were used to quantify the degree of co-localization between fluorophores. The Pearson's correlation coefficients for Superior LysoProbe I–IV and LysoTracker were high. Additionally, the overlap coefficients of the two fluorescence patterns were also relatively high (Table S1).

Bottom Line: The use of Superior LysoProbes facilitates the direct visualization of the lysosomal response to lobaplatin elicited in human chloangiocarcinoma (CCA) RBE cells, using confocal laser scanning microscopy.Additionally, we have characterized the role of lysosomes in autophagy, the correlation between lysosome function and microtubule strength, and the alteration of lysosomal morphology during apoptosis.Our findings indicate that Superior LysoProbes offer numerous advantages over previous reagents to examine the intracellular activities of lysosomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Michigan Technological University, Houghton, MI 49931.

ABSTRACT
Intracellular pH plays an important role in the response to cancer invasion. We have designed and synthesized a series of new fluorescent probes (Superior LysoProbes) with the capacity to label acidic organelles and monitor lysosomal pH. Unlike commercially available fluorescent dyes, Superior LysoProbes are lysosome-specific and are highly stable. The use of Superior LysoProbes facilitates the direct visualization of the lysosomal response to lobaplatin elicited in human chloangiocarcinoma (CCA) RBE cells, using confocal laser scanning microscopy. Additionally, we have characterized the role of lysosomes in autophagy, the correlation between lysosome function and microtubule strength, and the alteration of lysosomal morphology during apoptosis. Our findings indicate that Superior LysoProbes offer numerous advantages over previous reagents to examine the intracellular activities of lysosomes.

No MeSH data available.


Related in: MedlinePlus