Three-dimensional single-cell imaging for the analysis of RNA and protein expression in intact tumour biopsies

Nobuyuki Tanaka*, Shigeaki Kanatani, Dagmara Kaczynska, Keishiro Fukumoto, Lauri Louhivuori, Tomohiro Mizutani, Oded Kopper, Pauliina Kronqvist, Stephanie Robertson, Claes Lindh, Lorand Kis, Robin Pronk, Naoya Niwa, Kazuhiro Matsumoto, Mototsugu Oya, Ayako Miyakawa, Anna Falk, Johan Hartman, Cecilia Sahlgren, Hans CleversPer Uhlén

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

17 Citations (Scopus)


Microscopy analysis of tumour samples is commonly performed on fixed, thinly sectioned and protein-labelled tissues. However, these examinations do not reveal the intricate three-dimensional structures of tumours, nor enable the detection of aberrant transcripts. Here, we report a method, which we name DIIFCO (for diagnosing in situ immunofluorescence-labelled cleared oncosamples), for the multimodal volumetric imaging of RNAs and proteins in intact tumour volumes and organoids. We used DIIFCO to spatially profile the expression of diverse coding RNAs and non-coding RNAs at the single-cell resolution in a variety of cancer tissues. Quantitative single-cell analysis revealed spatial niches of cancer stem-like cells, and showed that the niches were present at a higher density in triple-negative breast cancer tissue. The improved molecular phenotyping and histopathological diagnosis of cancers may lead to new insights into the biology of tumours of patients.

Original languageEnglish
Pages (from-to)875-888
Number of pages14
JournalNature Biomedical Engineering
Issue number9
Publication statusPublished - 2020
MoE publication typeA1 Journal article-refereed


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