Modular low-light microscope for imaging cellular bioluminescence and radioluminescence

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Low-light microscopy is crucial for imaging light-sensitive or dim biological signals.
• Existing methods face limitations with phototoxicity and sensitivity for certain applications.
• Emerging applications require advanced techniques for non-perturbing cellular imaging.

Purpose of the Study:

• To provide a protocol for building a modular low-light microscope.
• To detail imaging protocols for radioluminescence, bioluminescence, and low-excitation fluorescence.
• To enable high-resolution imaging of small-molecule transport in single cells.

Main Methods:

• Construction of a modular microscope using standard optomechanical components.
• Coupling of two microscope objective lenses back-to-back.
• Utilizing a scintillator crystal for radioluminescence detection and image reconstruction software.

Main Results:

• Successful imaging of radioluminescence, bioluminescence, and low-excitation fluorescence signals.
• Demonstration of radioluminescence microscopy for studying small-molecule transport in single cells.
• High-resolution image reconstruction of radiotracer distribution.

Conclusions:

• The developed modular low-light microscope facilitates sensitive imaging without phototoxicity or photobleaching.
• Radiancoluminescence microscopy offers a novel method for studying cellular processes non-invasively.
• Parallel functional studies of cell heterogeneity at the single-cell level are achievable.