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

The fluorescence intracellular localization of Superior LysoProbe (IV) (1 μM) was examined in an additional three cancer cell lines: gastric cancer-derived HGC-27 cells, human colon cancer-derived CW-2 cells, and human breast cancer MCF-7 cell lines.The intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to LysoTracker Green (2 μM) in human colon cancer-derived CW-2 cells, gastric cancer-derived HGC-27 cells, and human breast cancer MCF-7 cell lines is shown. Superior LysoProbe (IV) (1 μM) 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). The cells were imaged on a confocal 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

f3: The fluorescence intracellular localization of Superior LysoProbe (IV) (1 μM) was examined in an additional three cancer cell lines: gastric cancer-derived HGC-27 cells, human colon cancer-derived CW-2 cells, and human breast cancer MCF-7 cell lines.The intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to LysoTracker Green (2 μM) in human colon cancer-derived CW-2 cells, gastric cancer-derived HGC-27 cells, and human breast cancer MCF-7 cell lines is shown. Superior LysoProbe (IV) (1 μM) 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). The cells were imaged on a confocal laser-scanning fluorescent microscope (Olympus) using a 60 × oil immersion objective lens.

Mentions: The fluorescence intracellular localization of Superior LysoProbes was further examined in three additional cancer cell lines: gastric cancer-derived HGC-27 cells, human colon cancer-derived CW-2 cells, and human breast cancer MCF-7 cells. As shown in Fig. 3, the fluorescence patterns of Superior LysoProbe IV and LysoTracker overlapped exactly, indicating lysosomal localization for Superior LysoProbe IV, and confirming that Superior LysoProbes localized in lysosomes in different cancer cell lines.


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)

The fluorescence intracellular localization of Superior LysoProbe (IV) (1 μM) was examined in an additional three cancer cell lines: gastric cancer-derived HGC-27 cells, human colon cancer-derived CW-2 cells, and human breast cancer MCF-7 cell lines.The intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to LysoTracker Green (2 μM) in human colon cancer-derived CW-2 cells, gastric cancer-derived HGC-27 cells, and human breast cancer MCF-7 cell lines is shown. Superior LysoProbe (IV) (1 μM) 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). The cells were imaged on a confocal 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

f3: The fluorescence intracellular localization of Superior LysoProbe (IV) (1 μM) was examined in an additional three cancer cell lines: gastric cancer-derived HGC-27 cells, human colon cancer-derived CW-2 cells, and human breast cancer MCF-7 cell lines.The intracellular distribution of Superior LysoProbe (IV) (1 μM) compared to LysoTracker Green (2 μM) in human colon cancer-derived CW-2 cells, gastric cancer-derived HGC-27 cells, and human breast cancer MCF-7 cell lines is shown. Superior LysoProbe (IV) (1 μM) 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). The cells were imaged on a confocal laser-scanning fluorescent microscope (Olympus) using a 60 × oil immersion objective lens.
Mentions: The fluorescence intracellular localization of Superior LysoProbes was further examined in three additional cancer cell lines: gastric cancer-derived HGC-27 cells, human colon cancer-derived CW-2 cells, and human breast cancer MCF-7 cells. As shown in Fig. 3, the fluorescence patterns of Superior LysoProbe IV and LysoTracker overlapped exactly, indicating lysosomal localization for Superior LysoProbe IV, and confirming that Superior LysoProbes localized in lysosomes in different cancer cell lines.

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