Multimodal illumination for quantitative transmitted-light microscopy

Area of Science:

• Microscopy
• Optical Imaging
• Biotechnology

Background:

• Advanced microscopy techniques like quantitative phase imaging are often limited to specialized, expensive equipment.
• Integrating diverse illumination modes (bright field, dark field, fluorescence, DPC) into a single, accessible module is challenging.

Purpose of the Study:

• To introduce the Compact Open-Source Multimodal Illumination Cluster (COSMIC) as a versatile and cost-effective illumination module.
• To enable multimodal and quantitative phase imaging on both commercial and open-source microscopy platforms.
• To provide a practical method for optimizing illuminator positioning.

Main Methods:

• Construction of the COSMIC module using low-cost, readily available components.
• Integration with µManager software for control and system compatibility.
• Demonstration of multimodal imaging on live cell cocultures and Caenorhabditis elegans.
• Validation and calibration of quantitative differential phase contrast (qDPC) using a phase target.

Main Results:

• COSMIC successfully enabled bright field, dark field, diascopic fluorescence, and differential phase contrast (DPC) imaging.
• Quantitative phase imaging (QPI) was achieved from DPC measurements on live specimens.
• Validated quantitative DPC (qDPC) across multiple microscope systems.
• Introduced a practical method for axial illuminator optimization.

Conclusions:

• COSMIC offers an accessible solution for multimodal and quantitative phase imaging.
• The module's low cost and ease of integration make advanced microscopy more widely available.
• COSMIC supports both research-grade and resource-constrained microscopy environments.