DNA‐Barcoded Fluorescence Microscopy for Spatial Omics

Innovation in genomics, transcriptomics, and proteomics research has created a plethora of state‐of‐the‐art techniques such as nucleic acid sequencing and mass‐spectrometry‐based proteomics with paramount impact in the life sciences. While current approaches yield quantitative abundance analysis of biomolecules on an almost routine basis, coupling this high content to spatial information in a single cell and tissue context is challenging. Here, current implementations of spatial omics are discussed and recent developments in the field of DNA‐barcoded fluorescence microscopy are reviewed. Light is shed on the potential of DNA‐based imaging techniques to provide a comprehensive toolbox for spatial genomics and transcriptomics and discuss current challenges, which need to be overcome on the way to spatial proteomics using high‐resolution fluorescence microscopy.

[1]  Suliana Manley,et al.  Waveguide-PAINT offers an open platform for large field-of-view super-resolution imaging , 2019, Nature Communications.

[2]  Guillaume Charras,et al.  Automating multimodal microscopy with NanoJ-Fluidics , 2018, bioRxiv.

[3]  P. Schwille,et al.  Flat-top TIRF illumination boosts DNA-PAINT imaging and quantification , 2019, Nature Communications.

[4]  F. Simmel,et al.  Isothermal assembly of DNA origami structures using denaturing agents. , 2008, Journal of the American Chemical Society.

[5]  Guo-Cheng Yuan,et al.  Transcriptome-scale super-resolved imaging in tissues by RNA seqFISH+ , 2019, Nature.

[6]  S. Hell,et al.  Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. , 1994, Optics letters.

[7]  Maximilian T. Strauss,et al.  Site-Specific Labeling of Affimers for DNA-PAINT Microscopy. , 2018, Angewandte Chemie.

[8]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[9]  Peng Yin,et al.  Optical visualisation of individual biomolecules in densely packed clusters , 2016 .

[10]  Timur Zhiyentayev,et al.  Single-cell in situ RNA profiling by sequential hybridization , 2014, Nature Methods.

[11]  Rithy K. Roth,et al.  Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays , 2000, Nature Biotechnology.

[12]  Emma Lundberg,et al.  Spatial proteomics: a powerful discovery tool for cell biology , 2019, Nature Reviews Molecular Cell Biology.

[13]  Ralf Jungmann,et al.  Up to 100-fold speedup and multiplexing in optimized DNA-PAINT , 2020, Nature Methods.

[14]  Ewert Bengtsson,et al.  Sequential immunofluorescence staining and image analysis for detection of large numbers of antigens in individual cell nuclei. , 2002, Cytometry.

[15]  Jean-Marie Rouillard,et al.  Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes , 2012, Proceedings of the National Academy of Sciences.

[16]  Mark Bates,et al.  Super-resolution fluorescence microscopy. , 2009, Annual review of biochemistry.

[17]  Nicholas A. Sinnott-Armstrong,et al.  Super-resolution chromatin tracing reveals domains and cooperative interactions in single cells , 2018, Science.

[18]  Chee-Huat Linus Eng,et al.  Profiling the transcriptome by RNA SPOTs , 2018 .

[19]  F. Simmel,et al.  Single-molecule kinetics and super-resolution microscopy by fluorescence imaging of transient binding on DNA origami. , 2010, Nano letters.

[20]  Salil S. Bhate,et al.  Deep Profiling of Mouse Splenic Architecture with CODEX Multiplexed Imaging , 2017, Cell.

[21]  Peng Yin,et al.  DNA-barcoded labeling probes for highly multiplexed Exchange-PAINT imaging† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6sc05420j Click here for additional data file. , 2017, Chemical science.

[22]  Peng Yin,et al.  Single-molecule super-resolution imaging of chromosomes and in situ haplotype visualization using Oligopaint FISH probes , 2015, Nature Communications.

[23]  P. Rothemund Folding DNA to create nanoscale shapes and patterns , 2006, Nature.

[24]  Sandro Santagata,et al.  Highly multiplexed immunofluorescence imaging of human tissues and tumors using t-CyCIF and conventional optical microscopes , 2018, eLife.

[25]  Bernd Bodenmiller,et al.  Multiplexed Epitope-Based Tissue Imaging for Discovery and Healthcare Applications. , 2016, Cell systems.

[26]  Peng Yin,et al.  Universal Super-Resolution Multiplexing by DNA Exchange. , 2017, Angewandte Chemie.

[27]  Maximilian T. Strauss,et al.  An order of magnitude faster DNA-PAINT imaging by optimized sequence design and buffer conditions , 2019, Nature Methods.

[28]  Johannes B. Woehrstein,et al.  Multiplexed 3D Cellular Super-Resolution Imaging with DNA-PAINT and Exchange-PAINT , 2014, Nature Methods.

[29]  Samir Kumar-Singh,et al.  Antibody Elution Method for Multiple Immunohistochemistry on Primary Antibodies Raised in the Same Species and of the Same Subtype , 2009, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[30]  G. von Heijne,et al.  Tissue-based map of the human proteome , 2015, Science.

[31]  Andrew D Ellington,et al.  Aptamers as potential tools for super-resolution microscopy , 2012, Nature Methods.

[32]  F. Simmel,et al.  Determination of DNA melting temperatures in diffusion-generated chemical gradients. , 2007, Analytical chemistry.

[33]  Chenglong Xia,et al.  Spatial transcriptome profiling by MERFISH reveals subcellular RNA compartmentalization and cell cycle-dependent gene expression , 2019, Proceedings of the National Academy of Sciences.

[34]  Marie-Lena I. E. Harwardt,et al.  Single-Molecule Super-Resolution Microscopy Reveals Heteromeric Complexes of MET and EGFR upon Ligand Activation , 2020, International journal of molecular sciences.

[35]  H. Ewers,et al.  A simple, versatile method for GFP-based super-resolution microscopy via nanobodies , 2012, Nature Methods.

[36]  Qing Li,et al.  Highly multiplexed single-cell analysis of formalin-fixed, paraffin-embedded cancer tissue , 2013, Proceedings of the National Academy of Sciences.

[37]  Peng Yin,et al.  Submicrometre geometrically encoded fluorescent barcodes self-assembled from DNA. , 2012, Nature chemistry.

[38]  Sean C. Bendall,et al.  Multiplexed ion beam imaging of human breast tumors , 2014, Nature Medicine.

[39]  Peter K. Sorger,et al.  Highly multiplexed imaging of single cells using a high-throughput cyclic immunofluorescence method , 2015, Nature Communications.

[40]  J. Elf,et al.  Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes , 2016, Science.

[41]  George M. Church,et al.  Highly Multiplexed Subcellular RNA Sequencing in Situ , 2014, Science.

[42]  M. Heilemann,et al.  Single-Molecule Localization Microscopy in Eukaryotes. , 2017, Chemical reviews.

[43]  Ruedi Aebersold,et al.  Mass-spectrometric exploration of proteome structure and function , 2016, Nature.

[44]  Amina A. Qutub,et al.  Multiplexed in situ immunofluorescence using dynamic DNA complexes. , 2012, Angewandte Chemie.

[45]  Erik K. Malm,et al.  A Human Protein Atlas for Normal and Cancer Tissues Based on Antibody Proteomics* , 2005, Molecular & Cellular Proteomics.

[46]  S. Hell,et al.  Fluorescence nanoscopy in cell biology , 2017, Nature Reviews Molecular Cell Biology.

[47]  Mike Heilemann,et al.  Automated highly multiplexed super-resolution imaging of protein nano-architecture in cells and tissues , 2020, Nature Communications.

[48]  Ryan M. Schweller,et al.  Programming in Situ Immunofluorescence Intensities through Interchangeable Reactions of Dynamic DNA Complexes , 2012, Chembiochem : a European journal of chemical biology.

[49]  Maximilian T. Strauss,et al.  124-Color Super-resolution Imaging by Engineering DNA-PAINT Blinking Kinetics , 2019, Nano letters (Print).

[50]  Michael R. Diehl,et al.  Configuring robust DNA strand displacement reactions for in situ molecular analyses , 2011, Nucleic acids research.

[51]  Ahmet F. Coskun,et al.  Dense transcript profiling in single cells by image correlation decoding , 2016, Nature Methods.

[52]  Maximilian T. Strauss,et al.  Modified aptamers enable quantitative sub-10-nm cellular DNA-PAINT imaging , 2018, Nature Methods.

[53]  Long Cai,et al.  Single cell systems biology by super-resolution imaging and combinatorial labeling , 2012, Nature Methods.

[54]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[55]  Yu Wang,et al.  Immuno-SABER enables highly multiplexed and amplified protein imaging in tissues , 2019, Nature Biotechnology.

[56]  Ulrike Endesfelder,et al.  A peptide tag-specific nanobody enables high-quality labeling for dSTORM imaging , 2018, Nature Communications.

[57]  J. Buhmann,et al.  Highly multiplexed imaging of tumor tissues with subcellular resolution by mass cytometry , 2014, Nature Methods.

[58]  L. Cai,et al.  In Situ Transcription Profiling of Single Cells Reveals Spatial Organization of Cells in the Mouse Hippocampus , 2016, Neuron.

[59]  Christian Schlötterer,et al.  Gene expression profiling by massively parallel sequencing. , 2007, Genome research.

[60]  X. Zhuang,et al.  Spatially resolved, highly multiplexed RNA profiling in single cells , 2015, Science.

[61]  A. Turberfield,et al.  A DNA-fuelled molecular machine made of DNA , 2022 .

[62]  Maximilian T. Strauss,et al.  Super-resolution microscopy with DNA-PAINT , 2017, Nature Protocols.

[63]  Anthony J. Manzo,et al.  Do-it-yourself guide: how to use the modern single-molecule toolkit , 2008, Nature Methods.

[64]  Pavel Vesely,et al.  Handbook of Biological Confocal Microscopy, 3rd ed. By James B. Pawley, Editor. Springer Science + Business Media, LLC, New York (2006). ISBN 10: 0‐387‐25921‐X; ISBN 13: 987‐0387‐25921‐5; hardback; 28 + 985 pages , 2007 .