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In this Protocol, Barna et al. describe how to use VividSTORM software to correlate the cellular information obtained from confocal imaging with single-molecule localization information obtained from superresolution STORM imaging.
This protocol describes the use of fluorescently quenched activity-based probes (qABPs) that target cysteine cathepsins. This approach allows for tracking of protease activity in tissue samples, which can be used for the detection of tumor resection margins.
This Protocol describes the CRIS-PITCh and TAL-PITCh systems for MMEJ-based gene targeting using CRISPR-Cas or TALENs. The approach may be particularly useful in systems where HR- or NHEJ-mediated targeting is inefficient.
Metal-organic frameworks (MOFs) are porous, crystalline materials with well-defined structures that can be used in many applications, from gas storage to catalysis and drug storage. This protocol is for the preparation of the MOF NU-1000.
Viable circulating tumor cells are isolated using easily fabricated microfluidic devices. The hydrodynamic forces present in curvilinear microfluidic channels separate cells on the basis of size.
This protocol describes assays for assessing heart rate fluctuations in mice or tissue isolated from mice, including by using in vivo telemetry and in vitro electrophysiology of intact sinoatrial network preparations or isolated single sinoatrial node cells.
This protocol for enhancer scanning to locate regulatory regions in genomic loci enables the identification of candidate functional SNPs within a locus during functional analysis of genome-wide association studies.
This protocol describes a Cre-loxP method to measure uptake of extracellular vesicles (EVs). Uptake of EVs released from cells that express Cre recombinase is detected in reporter cell lines as a switch from DsRed to eGFP expression.
This virus-free approach for mRNA knockdown in vivo uses the siRNA carrier peptide protamine, chemically coupled to a cell surface receptor internalizing antibodies via sulfo-SMCC, to provide protection and guidance for the targeted application of siRNA.
In this protocol, the construction and use of an operationally simple photochemical microreactor for visible light gas-liquid photoredox catalysis is described. The procedure includes details on how to set up and use the microreactor.
It is important to check whether an expressed membrane protein is successfully secreted to the cell surface. Extracellular protease digestion of intact cells followed by immunoblotting is easy and informative, but it requires an often-omitted control.
Live imaging of regeneration of adult zebrafish tissues has been hampered by the inability to continuously anaesthetize fish for longer than 2 h. Xu et al. describe an intubation-based approach that extends that period to 2 d.
This protocol uses micropatterned co-cultures comprising 2D islands of primary human hepatocytes surrounded by supportive fibroblast cells to model liver infection by the hepatitis B and C viruses or Plasmodium pathogens in vitro.
Having demonstrated that brain endothelial cells cross-present parasite antigen in mouse experimental cerebral malaria, the authors present ex vivo and in vitro protocols to measure this antigen presentation, generalizable to other disease contexts.
This protocol describes use of the Exomiser suite, a collection of algorithms that allow for prioritization of genes and variants from exome sequencing data for disease-gene discovery.
Imaging FCS and FCCS generate high-resolution quantitative data over large numbers of pixels that can be used to measure a range of molecular characteristics, including concentration, diffusion rates and interactions.