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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

Confocal laser-scanning fluorescent images of Superior LysoProbe (IV) in HeLa cells.Superior LysoProbe IV (1 μM, red) was incubated with cells in non-FBS DMEM media for 15 min., and then counterstained with LysoTracker (2 μM, weak green fluorescence), Hoechst 33342 (1 μg/mL, blue), and followed by 48 h incubation. All images were acquired using a 60 × oil immersion objective lens.
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f6: Confocal laser-scanning fluorescent images of Superior LysoProbe (IV) in HeLa cells.Superior LysoProbe IV (1 μM, red) was incubated with cells in non-FBS DMEM media for 15 min., and then counterstained with LysoTracker (2 μM, weak green fluorescence), Hoechst 33342 (1 μg/mL, blue), and followed by 48 h incubation. All images were acquired using a 60 × oil immersion objective lens.

Mentions: We performed long-term retention studies following cell loading with fluorescent probes to assess the extent of intracellular retention. For these studies, Superior LysoProbe and LysoTracker Green were incubated with HeLa cells for 48 h and their fluorescent changes evaluated. Within 2 hours, HeLa cells loaded with LysoTracker Green displayed no observable fluorescence. Conversely, there was still significant fluorescence in HeLa cells loaded with Superior LysoProbe at 48 h, and the intracellular staining pattern remained punctate (images shown in Fig. 6), suggesting significant probe retention in the lysosome.


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)

Confocal laser-scanning fluorescent images of Superior LysoProbe (IV) in HeLa cells.Superior LysoProbe IV (1 μM, red) was incubated with cells in non-FBS DMEM media for 15 min., and then counterstained with LysoTracker (2 μM, weak green fluorescence), Hoechst 33342 (1 μg/mL, blue), and followed by 48 h incubation. All images were acquired 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

f6: Confocal laser-scanning fluorescent images of Superior LysoProbe (IV) in HeLa cells.Superior LysoProbe IV (1 μM, red) was incubated with cells in non-FBS DMEM media for 15 min., and then counterstained with LysoTracker (2 μM, weak green fluorescence), Hoechst 33342 (1 μg/mL, blue), and followed by 48 h incubation. All images were acquired using a 60 × oil immersion objective lens.
Mentions: We performed long-term retention studies following cell loading with fluorescent probes to assess the extent of intracellular retention. For these studies, Superior LysoProbe and LysoTracker Green were incubated with HeLa cells for 48 h and their fluorescent changes evaluated. Within 2 hours, HeLa cells loaded with LysoTracker Green displayed no observable fluorescence. Conversely, there was still significant fluorescence in HeLa cells loaded with Superior LysoProbe at 48 h, and the intracellular staining pattern remained punctate (images shown in Fig. 6), suggesting significant probe retention in the lysosome.

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