Spinning disk confocal microscopy with a 25 Megapixel Camera

Area of Science:

• Biomedical Imaging
• Microscopy Technology
• Optical Engineering

Background:

• Spinning disk confocal microscopy offers fast optical sectioning and low phototoxicity.
• High hardware costs limit accessibility of advanced spinning disk confocal microscopy systems.

Purpose of the Study:

• To develop a low-cost spinning disk confocal microscopy system.
• To demonstrate the feasibility of using affordable CMOS cameras in spinning disk microscopy.
• To provide an open-source and scalable solution for advanced biological imaging.

Main Methods:

• Utilized a 25-megapixel machine vision CMOS camera (Sony IMX540, FLIR Blackfly S).
• Designed and constructed a custom spinning disk with 18 μm pinholes.
• Integrated the camera and disk with an optical chopper motor and Micro-Manager software.

Main Results:

• Achieved high-resolution 3D imaging of biological samples (mouse brain slices, rat testis).
• Verified performance with high quantum efficiency (69% at 525 nm) and low read noise (2.31 electrons) CMOS sensor.
• Demonstrated stable, near-telecentric imaging capabilities.

Conclusions:

• Affordable CMOS sensors can effectively replace sCMOS cameras in spinning disk microscopy.
• The developed system offers a cost-effective alternative for advanced imaging.
• The open-source nature promotes scalability and wider adoption in research.