Nucleic acid and protein mass mapping by live-cell deep-ultraviolet microscopy

Area of Science:

• Biophysics
• Cell Biology
• Microscopy

Background:

• Traditional microscopy techniques often require cell labeling, which can introduce artifacts or alter cellular behavior.
• Understanding cellular processes like mitosis and motility requires high-resolution imaging with minimal disruption.

Purpose of the Study:

• To develop and validate a deep-ultraviolet (UV) microscopy technique for imaging live cells.
• To assess the potential of deep-UV microscopy for mapping cellular mass and fluorescence in unlabeled cells.
• To evaluate the toxicity and temporal limits of deep-UV microscopy in cultured human and mouse cells.

Main Methods:

• Development of a deep-ultraviolet (UV) microscope operating at 280 nm.
• Imaging of cell mitosis and motility in cultured human and mouse cells.
• Application of computational methods to convert pixel intensity to mass estimates (nucleic acid, protein) and fluorescence yield.

Main Results:

• Successful imaging of cell mitosis and motility for 45 minutes with minimal UV-induced toxicity.
• Extended imaging up to 6 hours before significant cell death onset.
• Generation of detailed maps of nucleic acid mass, protein mass, and fluorescence yield in unlabeled cells.

Conclusions:

• Deep-UV microscopy offers a non-invasive method for studying dynamic cellular processes.
• The technique enables quantitative mapping of cellular components without labeling.
• This approach holds promise for advancing live-cell imaging and cell biology research.