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High-speed, multi-modal, label-free imaging of pathological slices with a Bessel beam
Chi Liu1,2, Zhifeng Zhao3,2, Cheng Jin1
1State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instrument, Tsinghua University, Beijing 100084, China.
Abstract:
Optical imaging of stained pathological slices has become the gold standard for disease diagnosis. However, the procedure of sample preparation is complex and time-consuming. Multiphoton microscopy (MPM) is promising for label-free imaging, but the imaging speed is limited, especially for whole slice imaging. Here we propose a high-speed, multi-modal, label-free MPM by Bessel scan-based strip mosaicking. With a Bessel beam for excitation, the extended depth-of-focus not only enables full axial information acquisition at once, but also alleviates the demanding requirement of sample alignment. With the strip mosaicking protocol, we can save the time of frequent sample transferring. Besides, we add a closely-attached reflection mirror under the sample for enhancing epi-detection signals, and employ circularly polarized beams for recording comprehensive information. We demonstrate its application in multi-modal, label-free imaging of human gastric cancer slices and liver cancer slices, and show its potential in rapid disease diagnosis.
Insights
We developed a fast, label-free multiphoton microscopy (MPM) technique using Bessel beams and strip mosaicking. This method enhances imaging speed for pathological slices, aiding rapid disease diagnosis.
Area of Science:
- Biomedical Optics
- Pathology Imaging
- Microscopy Technology
Background:
- Optical imaging of stained pathological slices is standard for disease diagnosis but involves complex, time-consuming sample preparation.
- Multiphoton microscopy (MPM) offers label-free imaging but suffers from slow speeds, particularly for whole-slice analysis.
Purpose of the Study:
- To develop a high-speed, multi-modal, label-free MPM technique for efficient pathological slice imaging.
- To overcome the limitations of traditional staining methods and slow MPM imaging speeds.
Main Methods:
- Utilized Bessel beam excitation for extended depth-of-focus and reduced alignment sensitivity.
- Implemented a strip mosaicking protocol to accelerate whole-slice imaging and minimize sample handling.
- Incorporated a reflection mirror for enhanced epi-detection signals and circularly polarized beams for comprehensive data acquisition.
Main Results:
- Demonstrated high-speed, multi-modal, label-free imaging of human gastric and liver cancer slices.
- Achieved efficient axial information acquisition and reduced sample alignment requirements.
- Showcased the potential for rapid, label-free disease diagnosis through enhanced imaging speed and signal detection.
Conclusions:
- The proposed Bessel scan-based strip mosaicking MPM is a significant advancement for label-free pathological imaging.
- This technique offers a faster and more efficient alternative to traditional methods, with potential for improved diagnostic workflows.
- The multi-modal capabilities provide comprehensive information for accurate disease assessment.
