Pre-Label-Free Three-Dimensional Imaging of Cellular Bottom Topography with a Charge-Lock Electrochemiluminescence...

Area of Science:

• Cell Biology
• Biophysics
• Microscopy

Background:

• Cellular bottom structure influences cell mobility.
• Studying cellular topography is challenging due to narrow spaces and potential damage from current microscopy techniques.
• Panoramic mapping of cellular bottom topography variations is difficult.

Purpose of the Study:

• To develop a novel microscopy technique for high-resolution mapping of cellular bottom topography.
• To overcome limitations of current ultra-high-resolution optical microscopy, such as cellular damage and off-target luminophore effects.
• To identify cell-matrix gap distance heterogeneity and its implications.

Main Methods:

• Charge-lock spatiotemporal transformation electrochemiluminescence (CL-STTECL) microscopy.
• Utilizing charge exclusion between cationic luminophores and a proton film to "lock" electrochemiluminescence (ECL).
• Employing amine co-reactants to "unlock" the ECL process, enabling gap distance-dependent emission.

Main Results:

• Achieved axial spatial resolution of approximately 10 nm for cell-matrix gap distance.
• Demonstrated changes in cellular bottom topography under various external stimulations.
• Uncovered panoramic feature heterogeneity between normal tissue cells and metastatic cancer cells.

Conclusions:

• CL-STTECL microscopy provides a novel approach for studying cellular bottom topography with unprecedented resolution.
• The technique can reveal topographical differences between normal and cancerous cells, suggesting potential for early cancer screening.
• This method offers a non-damaging way to map cellular structures at the nanoscale.