Phase and fluorescence imaging with a surprisingly simple microscope based on chromatic aberration

Area of Science:

• Microscopy
• Biophotonics
• Cell Biology

Background:

• Standard microscopy techniques often require bulky equipment and complex setups.
• Simultaneous phase contrast and fluorescence imaging can provide complementary information about cell morphology and function.
• Long-term cell culture imaging under physiological conditions is crucial for studying dynamic biological processes.

Purpose of the Study:

• To develop a simple, compact, and cost-effective microscope system.
• To enable simultaneous phase imaging and multi-color fluorescence imaging.
• To facilitate long-term cell culture imaging within a biological incubator.

Main Methods:

• A novel microscope design combining phase imaging and epi-fluorescence.
• Phase images reconstructed from an out-of-focus bright-field image.
• Wavelength-dependent focal plane shifting achieved through engineered chromatic aberration.
• No mechanical components required for modality switching.

Main Results:

• The system successfully integrates phase contrast and multi-color fluorescence imaging.
• Reproducible focal plane shifts achieved by changing imaging wavelength.
• Demonstrated long-term cell culture imaging within a biological incubator under physiological conditions.
• Achieved a field-of-view of 1.2 mm² with micrometer spatial resolution.

Conclusions:

• The proposed microscope is a compact, user-friendly, and affordable tool for biological research.
• Its ability to image in phase and fluorescence modes simultaneously within an incubator is a significant advantage.
• The system supports long-term observation of cellular dynamics under physiological conditions.