Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells.

Area of Science:

• Biophysics
• Cell Biology
• Optical Microscopy

Background:

• Quantitative phase imaging (QPI) provides label-free contrast for transparent specimens.
• Fluorescence microscopy offers molecular specificity but typically requires separate imaging channels.
• Simultaneous QPI and fluorescence imaging can enhance cell analysis but often involves complex setups.

Purpose of the Study:

• To present a new technique for simultaneous multimodal quantitative phase and fluorescence microscopy.
• To enable both quantitative phase imaging and molecular specificity using a single camera.
• To overcome limitations of previous approaches by allowing conventional fluorescence imaging.

Main Methods:

• Development of an interferometric multiplexing module externally positioned at the exit of an optical microscope.
• Integration of quantitative phase and fluorescence imaging modalities onto a single camera.
• Demonstration using fluorescent beads and live biological cells.

Main Results:

• Successful simultaneous acquisition of quantitative phase and fluorescence images.
• Demonstration of the technique's capability for imaging fluorescent beads.
• Validation of the method on live biological cells, showcasing its applicability.

Conclusions:

• The presented technique offers a streamlined approach for multimodal microscopy.
• It provides both label-free phase contrast and molecular specificity in a single acquisition.
• This method has potential applications in live-cell imaging and biological research.