3D MERFISH Imaging of Thick Tissues
Author Information
Author(s): Fang Rongxin, Halpern Aaron, Rahman Mohammed Mostafizur, Huang Zhengkai, Lei Zhiyun, Hell Sebastian J, Dulac Catherine, Zhuang Xiaowei
Primary Institution: Howard Hughes Medical Institute, Harvard University
Hypothesis
Can we extend the MERFISH method to enable 3D imaging of thick tissues up to 200 µm?
Conclusion
The study successfully demonstrates a method for 3D MERFISH imaging in thick tissues, allowing for high-quality spatial transcriptomics.
Supporting Evidence
- 3D MERFISH imaging allows for volumetric single-cell transcriptome imaging of thick-tissue samples.
- The method demonstrated high detection efficiency and accuracy in mouse brain tissue sections.
- Confocal microscopy was used to eliminate out-of-focus signals, improving image quality.
- Deep learning was applied to enhance the signal-to-noise ratio in images.
- The study provides a new tool for spatial genomics, broadening the scope of questions that can be addressed.
Takeaway
This study shows how to take pictures of tiny parts of thick tissues to see what genes are doing inside cells.
Methodology
The study used multiplexed error-robust fluorescence in situ hybridization (MERFISH) combined with deep learning and confocal microscopy to image thick tissue samples.
Limitations
The method may not be applicable to all types of thick tissues and requires further validation in different biological contexts.
Participant Demographics
Adult C57BL/6J male mice aged 7–9 weeks were used.
Digital Object Identifier (DOI)
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