A recent study by Ravian van Ineveld and Michiel Kleinnijenhuis et al. in Nature Biotechnology shows the development of a new method for determining molecular, spatial and morphological features of cells in 3D tissues.
The research group led by dr. Anne Rios (Princess Máxima Center for pediatric oncology, Oncode Institute) developed an eight-color, multispectral, Large-scale Single-cell Resolution 3D (mLSR-3D) imaging technology, accompanied by a deep learning–based image analysis software for the parallelized segmentation of large numbers of single cells in tissues (STAPL-3D). These methodologies revealed an unprecedented level of detail on the molecular, spatial and morphological features of cells in Wilms tumor and healthy human fetal kidney.
To classify the different cell types, the authors selected 5 cell type- and 3 general markers out of a list of 60 antibodies and dyes. QVQ’s anti-NCAM VHH molecule (product Q55) was one of these 5 cell type markers. Using the proprietary C-Direct tag at the C-terminus of the VHH, QVQ directionally labeled Q55 with either HiLyte Fluor 488 or HiLyte Fluor 555. Of these, Q55c-555 showed good signal-to-noise ratio, excellent tissue penetration and compatibility with linear spectral unmixing.
This study again emphasizes that VHHs serve well for 3D tissue imaging. Their small size and relatively high affinity ensure excellent tissue penetration and retention. By using direct fluorescently labeled VHH, mixtures of differentially labeled VHHs could provide a one-pot approach for the simultaneous staining of different markers.
This work by van Ineveld and Kleinnijenhuis et al. can be accessed via: https://www.nature.com/articles/s41587-021-00926-3.