Single plane illumination module and micro-capillary approach for a wide-field microscope.

Area of Science:

• Biomedical Imaging
• Microscopy Techniques
• Cell Biology

Background:

• Advanced microscopy is crucial for studying complex 3D cell cultures.
• Existing systems may lack adaptability for diverse biological samples and applications.
• Single Plane Illumination Microscopy (SPIM) offers optical sectioning capabilities.

Purpose of the Study:

• To present an adaptable SPIM module for inverted wide-field microscopes.
• To optimize the module for 3-dimensional cell cultures, such as multicellular tumor spheroids (MCTS).
• To demonstrate its utility in drug uptake, redox, and cell death studies.

Main Methods:

• Development of a SPIM excitation module for perpendicular illumination.
• Integration with an inverted wide-field microscope.
• Utilization of rectangular capillaries for sample holding and rotation.
• Synchronous adjustment of light sheet and objective lens.
• Incorporation of a microfluidic system for reagent delivery.

Main Results:

• Quantification of cytostatic drug (doxorubicin) uptake and degradation.
• Monitoring of cellular redox status using a glutathione sensor.
• Induction and visualization of cell necrosis via mitochondrial inhibition.
• Demonstration of the system's advantages over existing SPIM setups.

Conclusions:

• The described SPIM module provides a versatile and efficient platform for advanced 3D cell culture imaging.
• It facilitates detailed studies of drug effects and cellular responses at the microscale.
• The system's adaptability and integrated features offer significant advantages for biological research.