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Nature Protocols is an interactive online resource for all laboratory protocols relevant to biological and biomedical research. Nature Protocols includes a free-to-access community site, 'Protocols Network', containing protocols that are posted directly o



 



On-chip human microvasculature assay for visualization and quantification of tumor cell extravasation dynamics

2017-03-30

This protocol describes how to set up and use an in vitro model of the human microcirculation with the capability to recapitulate discrete steps of early metastatic seeding, including tumor cell arrest and transendothelial migration.



Analyzing trapped protein complexes by Virotrap and SFINX

2017-03-30

This protocol enables the study of protein complexes by trapping them in virus-like particles and thereby preserving cellular integrity. The Virotrap protocol is complemented by SFINX, an online data analysis tool for interactomics studies.



Long-read ChIA-PET for base-pair-resolution mapping of haplotype-specific chromatin interactions

2017-03-30

Li et al. provide a protocol for long-read ChIA-PET, a technique for mapping chromatin interactions. The longer paired-end tags, which are generated by tagmentation, provide sufficient coverage to determine haplotype-specific chromatin interactions at single-nucleotide resolution.



Diverse protocols for correlative super-resolution fluorescence imaging and electron microscopy of chemically fixed samples

2017-04-06

Four different techniques for preparing and acquiring super-resolution CLEM data sets on aldehyde-fixed specimens are provided: Tokuyasu cryosectioning; whole-cell mount; cell unroofing and platinum replication; and resin embedding and sectioning.



Antifungal drug testing by combining minimal inhibitory concentration testing with target identification by gas chromatography–mass spectrometry

2017-04-06

By combining minimal inhibitory concentration testing with GC–MS-based pathway analysis, it is possible to identify which enzymes in the ergosterol biosynthesis pathway are being targeted by potential new antifungal substances.



Preparation of molecularly imprinted polymers specific to glycoproteins, glycans and monosaccharides via boronate affinity controllable–oriented surface imprinting

2017-04-06

This protocol describes how to produce molecularly imprinted polymers, synthetic receptors that can be produced with antibody-like binding properties. They are easier and cheaper to produce than antibodies and lectins.



Strategic and practical guidelines for successful structured illumination microscopy

2017-04-13

This protocol describes the preparation of calibration bead slides, their use and additional strategies to reduce artifacts of structured illumination microscopy that will allow researchers to exploit the technique's full potential for biological applications.



Quantitative 3D structured illumination microscopy of nuclear structures

2017-04-13

This protocol describes how to prepare samples for labeling nuclei of cultured mammalian cells for 3D structured illumination microscopy of nuclear structures. Image acquisition, registration and downstream image analysis are also described.



Generation of multipotent induced cardiac progenitor cells from mouse fibroblasts and potency testing in ex vivo mouse embryos

2017-04-20

Expandable and multipotent induced cardiac progenitor cells (iCPCs) are generated from mouse adult fibroblasts using forced expression of Mesp1, Tbx5, Gata4, Nkx2.5 and Baf60c and activation of the Wnt and JAK/STAT signaling pathway. Furthermore, embryonic potency of iCPCs is tested ex vivo.



Modular low-light microscope for imaging cellular bioluminescence and radioluminescence

2017-04-20

Bioluminescence and radioluminescence are both dim phenomena that require a highly sensitive microscope for imaging at the cellular level. This protocol describes how to construct a modular low-light microscope for imaging these events.