Dual light-emitting diode-based multichannel microscopy for whole-slide multiplane, multispectral and phase imaging

Area of Science:

• Biomedical Engineering
• Optical Microscopy
• Image Processing

Background:

• Traditional whole-slide imaging (WSI) can be time-consuming and may not capture sufficient detail for complex biological samples.
• Multiplexed imaging and phase contrast are crucial for detailed cellular and tissue analysis.
• Current WSI techniques often require sequential acquisition of different focal planes or spectral bands, limiting speed and dynamic range.

Purpose of the Study:

• To develop a novel multichannel microscopy system for simultaneous multiplane, multispectral, and phase imaging in whole-slide imaging.
• To achieve high data throughput and rapid autofocusing for efficient biological sample analysis.
• To provide an open-source platform for advanced multiplexed and multimodal WSI.

Main Methods:

• Utilized trinocular heads to create 6 independent imaging channels for parallel data acquisition.
• Implemented a camera array for high-speed data throughput (approx. 1 gigapixel/second).
• Integrated near-infrared LEDs and a hot mirror for single-frame, rapid autofocusing and employed the transport-of-intensity equation for phase recovery.

Main Results:

• Successfully demonstrated simultaneous acquisition of 6 focal planes and 6 spectral bands for WSI.
• Achieved whole-slide phase imaging by acquiring images at 3 focal positions.
• Developed an open-source design capable of expanding to 15 imaging channels.

Conclusions:

• The developed multichannel microscopy platform offers a simplified solution for multiplexed fluorescence and multimodal WSI.
• The system enables rapid acquisition of focal stacks without z-scanning, facilitating fast 3D dynamic tracking of biological samples.
• This technology has the potential to significantly advance high-throughput biological imaging and diagnostics.