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Seeing nanoscale organization through a STED doughnut. Nanoscale membrane heterogeneity is investigated using a combination of super-resolution STED microscopy and fluorescence correlation spectroscopy.
Brodkorb et al. provide a standardized static in vitro protocol for the study of gastrointestinal food digestion and the analysis of digestion products.
Sonoporation is a minimally invasive gene delivery method mediated by ultrasound. This protocol describes step-by-step procedures for sonoporation in pigs and mice.
Zhang and Lyden describe a protocol for asymmetric-flow field-flow fractionation (AF4) to separate and characterize extracellular nanoparticles for investigation of their biogenesis, function and potential in molecular diagnostics and therapeutics.
This protocol describes how to measure nanoscale diffusion dynamics of proteins and lipids in cell membranes using STED–FCS. The protocol contains procedures for calibration, performing point and scanning STED–FCS measurements, and data analysis.
This protocol describes the experimental design and procedures for bioluminescence resonance energy transfer (BRET) imaging. The authors discuss choices of energy donors and acceptors, luminescent substrates, microscope setup and cameras.
Amide bond formation and aromatic nitro-group reductions are common reactions that are environmentally harmful. An alternative approach is to perform the reactions in water at ambient temperatures using small amounts of a designer surfactant.
This protocol describes the fabrication and practical applications of molybdenum disulfide (MoS2) nanopores. The procedure contains different methods for the transfer of monolayer MoS2, nanopore creation, and data acquisition and analysis.
This protocol describes how to perform nanoscale chemical imaging using tip-enhanced Raman spectroscopy (TERS). The procedure details the preparation of plasmonically active TERS probes, alignment of a TERS system, and various example procedures.
The human cochlea is a rich source of endogenous DNA. This protocol describes how to use a sandblaster to isolate the cochlea from ancient skeletal remains and prepare bone powder for downstream DNA extraction.
The particle-swarm optimization–enhanced thromboSeq pipeline enables RNA sequencing and cancer classification from blood platelet RNA, using self-learning and swarm intelligence–enhanced bioinformatics algorithms.
This protocol describes how to injure and trace dorsal column sensory and corticospinal axons and visualize axon degeneration and regeneration by two-photon microscopy. Axon density and regeneration are assessed with Imaris and ImageJ software.
Human pluripotent stem cells are differentiated first into enteric neural crest cells and then into functional enteric neurons using a defined culture system.
This protocol describes intracellular temperature imaging with a cationic linear fluorescent polymeric thermometer (FPT). Step-by-step procedures are provided for sensor calibration and imaging by fluorescence lifetime imaging microscopy (FLIM).