Development of a nanobody-based, slide-free approach for 3D-Histological analysis of the spatial tumor microenvironment using lightsheet imaging

Introduction

For histopathological analysis and subsequent diagnosis of diseased tissues, a rapid, accurate histopathological and genetic analysis of biopsied tissues is essential. However, traditional 2-dimensional slide-based histopathological analysis does not represent spatial tissue structures and molecular targets sufficiently. This limitation aggravates an accurate and spatial analysis of tissue, a detailed description of the tumor microenvironment, as well as patient stratification.

Proposed Solution

To overcome these limitations, I propose an innovative, nanobody-based 3D-histopathology approach using non-destructive volumetric lightsheet microscopy. This approach bears the potential to revolutionize the way pathological analysis of tumor samples is conducted.

Advantages of 3D-Histopathology

In comparison to conventional 2D-pathology, non-destructive 3D-histopathology allows:

1. Rapid slide-free histological imaging of an entire tissue sample.
2. Volumetric analysis of diagnostic relevant structures such as immune cells and vessels.
3. Improved spatial analysis of cell distribution relevant to studying the microenvironment.

Methodology

To achieve this, we will generate nanobodies for improved wholemount tissue staining of the immune system as well as tumor-relevant markers. By labeling nanobodies with cleavable fluorescent dyes using click chemistry and integrating linkers for stoichiometric and directed labeling, we will be able to perform several rounds of multiplex staining of the identical sample. This will provide cutting-edge diagnostic phenotyping of the specimen.

Non-Destructive Nature

Due to the non-destructive nature of the optical-sectioning methodology, the sample will remain available for downstream applications such as molecular diagnostics of the previously imaged specimen.

Conclusion

In summary, this innovative project holds great potential to revolutionize cancer histology by providing a spatial and detailed description of the tumor microenvironment, improving the diagnosis of tumor samples, and consequently enhancing patient stratification and outcome.