Versatile, do-it-yourself, low-cost spinning disk confocal microscope

Area of Science:

• Biotechnology
• Microscopy
• Optical Engineering

Background:

• Confocal microscopy is essential for 3D biological imaging but is often limited by high hardware costs.
• Existing confocal microscopy solutions are typically confined to specialized core facilities, restricting broader research access.

Purpose of the Study:

• To develop an inexpensive do-it-yourself (DIY) spinning disk confocal microscope (SDCM) module.
• To increase the accessibility of confocal microscopy for researchers by reducing hardware costs.

Main Methods:

• Fabrication of a spinning disk confocal microscope module using a commercially available chromium photomask.
• Integration of the DIY SDCM module onto a standard laser-illuminated epifluorescence microscope.
• Testing the SDCM module's performance across a broad wavelength range (approximately 400-800 nm).

Main Results:

• The DIY SDCM module demonstrated strong performance in various biological imaging applications.
• Successful imaging was achieved using conventional microscopy techniques, including immunofluorescence, small-molecule stains, and fluorescence in situ hybridization.
• The module also supported super-resolution microscopy methods such as single-molecule localization microscopy and expansion microscopy.

Conclusions:

• The developed low-cost DIY SDCM module significantly enhances accessibility to confocal microscopy.
• This accessible technology empowers researchers with advanced 3D imaging capabilities without prohibitive costs.
• The project provides comprehensive documentation to facilitate the adoption of this DIY confocal microscopy solution.