Whole-Cell Lysosome SMLM Imaging as Indicators for Functional Diagnostics with a Low-Phototoxic Spontaneously...

Area of Science:

• Cell Biology
• Microscopy
• Biochemistry

Background:

• Lysosomal morphology and pH dynamics are crucial for cellular function and disease states.
• Long-term imaging is vital for understanding lysosomal dynamics but is limited by phototoxicity.
• Single-molecule localization microscopy (SMLM) offers high-resolution insights but requires advanced probes.

Purpose of the Study:

• To develop a novel imaging probe for low-phototoxicity, long-term SMLM of lysosomes.
• To investigate lysosomal morphology and pH dynamics in response to various stimuli and drug treatments.
• To establish a robust platform for lysosomal functional diagnostics and drug screening.

Main Methods:

• Development of Aze-HMSiR, a spontaneously blinking silicon rhodamine probe with near-infrared excitation.
• Long-term (50 min) SMLM imaging of lysosomes using Aze-HMSiR.
• Assessment of lysosomal morphology, size, distribution, and lumen pH under physiological and pathological conditions, including drug treatments.

Main Results:

• Aze-HMSiR enabled low-phototoxicity, long-term SMLM imaging of lysosomal morphology and pH dynamics.
• Super-resolution imaging provided insights into lysosomal distribution, size, and lumen pH.
• Periplocoside reduced lysosomal size, while paclitaxel increased lysosomal pH and altered distribution in cancer drug screening.

Conclusions:

• Aze-HMSiR is a powerful tool for lysosomal functional diagnostics, offering super-resolution insights.
• The probe facilitates long-term monitoring of lysosomal dynamics under various perturbations.
• This technology supports drug screening and the study of lysosomal roles in disease, particularly cancer therapy.