Cervical duct cannulation in sheep for collection of afferent lymph dendritic cells from head tissues.

Abstract:

:Pseudo-afferent cervical lymph-duct cannulation in a sheep model allows large amounts of lymph cells to be collected under physiological conditions, carrying immune signaling information from the head tissues, including oro-nasal mucosae. Importantly, large quantities of dendritic cells (DCs) of several subtypes are obtained (up to 8 million per overnight collection), as well as many other trafficking leukocytes. The technique includes three steps: removal of all head lymph nodes on one side (2 h), catheterization of cervical lymph ducts after 2 months (2-3 h) and collection/purification of lymph-cell subsets (4 h). The approach is challenging (1 in 3 success rate) but fruitful, and can be used to study DC subsets under immunomodulation, in order to assess lymph-cell subset dynamic changes and antigen transportation from oro-nasal tissues. This protocol is directed to experienced postdoctoral researchers.

journal_name

Nat Protoc

journal_title

Nature protocols

authors

Schwartz-Cornil I,Epardaud M,Bonneau M

doi

10.1038/nprot.2006.147

subject

Has Abstract

pub_date

2006-01-01 00:00:00

pages

874-9

issue

2

eissn

1754-2189

issn

1750-2799

pii

nprot.2006.147

journal_volume

1

pub_type

杂志文章
  • Long-read sequencing data analysis for yeasts.

    abstract::Long-read sequencing technologies have become increasingly popular due to their strengths in resolving complex genomic regions. As a leading model organism with small genome size and great biotechnological importance, the budding yeast Saccharomyces cerevisiae has many isolates currently being sequenced with long read...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2018.025

    authors: Yue JX,Liti G

    更新日期:2018-06-01 00:00:00

  • Virtual karyotyping of pluripotent stem cells on the basis of their global gene expression profiles.

    abstract::The genomic instability of stem cells in culture, caused by their routine in vitro propagation or by their genetic manipulation, is deleterious both for their clinical application and for their use in basic research. Frequent evaluation of the genomic integrity of stem cells is thus required, and it is usually perform...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.051

    authors: Ben-David U,Mayshar Y,Benvenisty N

    更新日期:2013-05-01 00:00:00

  • Synthesis of alkyl- and aryl-amino-substituted anthraquinone derivatives by microwave-assisted copper(0)-catalyzed Ullmann coupling reactions.

    abstract::This protocol describes the efficient, generally applicable Ullmann coupling reaction of bromaminic acid with alkyl- or aryl-amines in phosphate buffer under microwave irradiation using elemental copper as a catalyst. The reaction leads to a number of biologically active compounds. As a prototypical example, the synth...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2010.63

    authors: Baqi Y,Müller CE

    更新日期:2010-05-01 00:00:00

  • Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes.

    abstract::Aptamers are single-stranded DNA or RNA molecules that can bind target molecules with high affinity and specificity. The conformation of an aptamer usually changes upon binding to its target analyte, and this property has been used in a wide variety of sensing applications, including detection based on fluorescence in...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.38

    authors: Liu J,Lu Y

    更新日期:2006-01-01 00:00:00

  • Solid-phase synthesis of short α-helices stabilized by the hydrogen bond surrogate approach.

    abstract::Stabilized α-helices and nonpeptidic helix mimetics have emerged as powerful molecular scaffolds for the discovery of protein-protein interaction inhibitors. Protein-protein interactions often involve large contact areas, which are often difficult for small molecules to target with high specificity. The hypothesis beh...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2010.146

    authors: Patgiri A,Menzenski MZ,Mahon AB,Arora PS

    更新日期:2010-11-01 00:00:00

  • Analyzing bacterial extracellular vesicles in human body fluids by orthogonal biophysical separation and biochemical characterization.

    abstract::Gram-negative and Gram-positive bacteria release a variety of membrane vesicles through different formation routes. Knowledge of the structure, molecular cargo and function of bacterial extracellular vesicles (BEVs) is primarily obtained from bacteria cultured in laboratory conditions. BEVs in human body fluids have b...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-019-0236-5

    authors: Tulkens J,De Wever O,Hendrix A

    更新日期:2020-01-01 00:00:00

  • High-throughput ballistic injection nanorheology to measure cell mechanics.

    abstract::High-throughput ballistic injection nanorheology is a method for the quantitative study of cell mechanics. Cell mechanics are measured by ballistic injection of submicron particles into the cytoplasm of living cells and tracking the spontaneous displacement of the particles at high spatial resolution. The trajectories...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2011.436

    authors: Wu PH,Hale CM,Chen WC,Lee JS,Tseng Y,Wirtz D

    更新日期:2012-01-05 00:00:00

  • The power of multiplexed functional analysis of genetic variants.

    abstract::New technologies have recently enabled saturation mutagenesis and functional analysis of nearly all possible variants of regulatory elements or proteins of interest in single experiments. Here we discuss the past, present, and future of such multiplexed (functional) assays for variant effects (MAVEs). MAVEs provide de...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.135

    authors: Gasperini M,Starita L,Shendure J

    更新日期:2016-10-01 00:00:00

  • Oxidative bisulfite sequencing of 5-methylcytosine and 5-hydroxymethylcytosine.

    abstract::To uncover the function of and interplay between the mammalian cytosine modifications 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), new techniques and advances in current technology are needed. To this end, we have developed oxidative bisulfite sequencing (oxBS-seq), which can quantitatively locate 5mC an...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.115

    authors: Booth MJ,Ost TW,Beraldi D,Bell NM,Branco MR,Reik W,Balasubramanian S

    更新日期:2013-10-01 00:00:00

  • A general purpose method for extracting RNA from Dictyostelium cells.

    abstract::Here we present a protocol for the extraction of RNA from Dictyostelium discoideum. Dictyostelium is a social amoeba that undergoes a basic developmental program, and therefore analysis of RNA levels over a time course is a commonly used technique. This procedure is similar to other guanidine thiocyanate-based methods...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.191

    authors: Pilcher KE,Gaudet P,Fey P,Kowal AS,Chisholm RL

    更新日期:2007-01-01 00:00:00

  • Fluorescence resonance energy transfer microscopy as demonstrated by measuring the activation of the serine/threonine kinase Akt.

    abstract::This protocol describes procedures for performing fluorescence resonance energy transfer (FRET) microscopy analysis by three different methods: acceptor photobleaching, sensitized emission and spectral imaging. We also discuss anisotropy and fluorescence lifetime imaging microscopy-based FRET techniques. By using the ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2012.147

    authors: Broussard JA,Rappaz B,Webb DJ,Brown CM

    更新日期:2013-02-01 00:00:00

  • Characterization of bacterial spore germination using phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers.

    abstract::This protocol describes a method combining phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers to characterize the germination of single bacterial spores. The characterization consists of the following steps: (i) loading heat-activated dormant spores into a temperature-controlled micros...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2011.307

    authors: Kong L,Zhang P,Wang G,Yu J,Setlow P,Li YQ

    更新日期:2011-05-01 00:00:00

  • Hydrodynamic size-based separation and characterization of protein aggregates from total cell lysates.

    abstract::Herein we describe a protocol that uses hollow-fiber flow field-flow fractionation (FFF) coupled with multiangle light scattering (MALS) for hydrodynamic size-based separation and characterization of complex protein aggregates. The fractionation method, which requires 1.5 h to run, was successfully modified from the a...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.009

    authors: Tanase M,Zolla V,Clement CC,Borghi F,Urbanska AM,Rodriguez-Navarro JA,Roda B,Zattoni A,Reschiglian P,Cuervo AM,Santambrogio L

    更新日期:2015-01-01 00:00:00

  • Use of luminescent bacteria for rapid screening and characterization of short cationic antimicrobial peptides synthesized on cellulose using peptide array technology.

    abstract::The increasing multi-resistance of pathogenic bacteria requires the development of novel classes of antibiotics. Antimicrobial host defense peptides represent one promising class. Here we describe a protocol for screening large numbers of peptides against any microbe of interest. Peptides synthesized on a cellulose su...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.203

    authors: Hilpert K,Hancock RE

    更新日期:2007-01-01 00:00:00

  • Micropatterning of living cells by laser-guided direct writing: application to fabrication of hepatic-endothelial sinusoid-like structures.

    abstract::Here, we describe a simple protocol for the design and construction of a laser-guided direct writing (LGDW) system able to micropattern the self-assembly of liver sinusoid-like structures with micrometer resolution in vitro. To the best of our knowledge, LGDW is the only technique able to pattern cells "on the fly" wi...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.386

    authors: Nahmias Y,Odde DJ

    更新日期:2006-01-01 00:00:00

  • Germline transgenesis in pigs by cytoplasmic microinjection of Sleeping Beauty transposons.

    abstract::The pig has emerged as an important large animal model in biomedical and pharmaceutical research. We describe a protocol for high-efficiency germline transgenesis and sustained transgene expression in pigs by using the Sleeping Beauty (SB) transposon system. The protocol is based on co-injection of a plasmid encoding ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2014.010

    authors: Ivics Z,Garrels W,Mátés L,Yau TY,Bashir S,Zidek V,Landa V,Geurts A,Pravenec M,Rülicke T,Kues WA,Izsvák Z

    更新日期:2014-04-01 00:00:00

  • In vivo rapid gene delivery into postmitotic neocortical neurons using iontoporation.

    abstract::This protocol describes a method for directing the expression of genes of interest into postmitotic neocortical neurons in vivo. Microinjection of a DNA plasmid-amphiphilic molecule mix into the neocortex followed by delivery of an ad hoc electric pulse protocol during the first few days of life in mice allows rapid, ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.001

    authors: De la Rossa A,Jabaudon D

    更新日期:2015-01-01 00:00:00

  • Long term non-invasive imaging of embryonic stem cells using reporter genes.

    abstract::Development of non-invasive and accurate methods to track cell fate after delivery will greatly expedite transition of embryonic stem (ES) cell therapy to the clinic. In this protocol, we describe the in vivo monitoring of stem cell survival, proliferation and migration using reporter genes. We established stable ES c...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2009.100

    authors: Sun N,Lee A,Wu JC

    更新日期:2009-01-01 00:00:00

  • Vibrational spectroscopic image analysis of biological material using multivariate curve resolution-alternating least squares (MCR-ALS).

    abstract::Raman and Fourier transform IR (FTIR) microspectroscopic images of biological material (tissue sections) contain detailed information about their chemical composition. The challenge lies in identifying changes in chemical composition, as well as locating and assigning these changes to different conditions (pathology, ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.008

    authors: Felten J,Hall H,Jaumot J,Tauler R,de Juan A,Gorzsás A

    更新日期:2015-02-01 00:00:00

  • Characterizing peptides in individual mammalian cells using mass spectrometry.

    abstract::Cell-to-cell chemical signaling plays multiple roles in coordinating the activity of the functional elements of an organism, with these elements ranging from a three-neuron reflex circuit to the entire animal. In recent years, single-cell mass spectrometry (MS) has enabled the discovery of cell-to-cell signaling molec...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.277

    authors: Rubakhin SS,Sweedler JV

    更新日期:2007-01-01 00:00:00

  • Synthesis and site-specific incorporation of a simple fluorescent pyrimidine.

    abstract::We describe procedures for the synthesis of a fluorescent pyrimidine analog and its site-specific incorporation into a DNA oligomer. The 5'-protected and 3'-activated nucleoside 4 is synthesized in three steps with an overall yield of 40%. Site-specific incorporation into a DNA oligomer occurs with greater than 88% co...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.464

    authors: Greco NJ,Tor Y

    更新日期:2007-01-01 00:00:00

  • Identifying and quantifying proteolytic events and the natural N terminome by terminal amine isotopic labeling of substrates.

    abstract::Analysis of the sequence and nature of protein N termini has many applications. Defining the termini of proteins for proteome annotation in the Human Proteome Project is of increasing importance. Terminomics analysis of protease cleavage sites in degradomics for substrate discovery is a key new application. Here we de...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2011.382

    authors: Kleifeld O,Doucet A,Prudova A,auf dem Keller U,Gioia M,Kizhakkedathu JN,Overall CM

    更新日期:2011-09-22 00:00:00

  • Isolation of DNA from small amounts of elephant ivory.

    abstract::This protocol describes a method for the extraction of DNA from elephant ivory. These techniques are being used to assign geographic origin to poached ivory by comparing the ivory genotype to a geographic-based gene frequency map, developed separately. The method has three components: ivory pulverization, decalcificat...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.318

    authors: Mailand C,Wasser SK

    更新日期:2007-01-01 00:00:00

  • A real-time fluorometric method for the simultaneous detection of cell death type and rate.

    abstract::Several cell death assays have been developed based on a single biochemical parameter such as caspase activation or plasma membrane permeabilization. Our fluorescent apoptosis/necrosis (FAN) assay directly measures cell death and distinguishes between caspase-dependent apoptosis and caspase-independent necrosis of cel...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.085

    authors: Grootjans S,Hassannia B,Delrue I,Goossens V,Wiernicki B,Dondelinger Y,Bertrand MJ,Krysko DV,Vuylsteke M,Vandenabeele P,Vanden Berghe T

    更新日期:2016-08-01 00:00:00

  • Analysis of neurotransmitter release mechanisms by photolysis of caged Ca²⁺ in an autaptic neuron culture system.

    abstract::Neurotransmitter release is triggered by membrane depolarization, Ca(2+) influx and Ca(2+) sensing by the release machinery, causing synaptic vesicle (SV) fusion with the plasma membrane. Interlinked is a complex membrane cycle in which vesicles are tethered to the release site, primed, fused and recycled. As many of ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2012.074

    authors: Burgalossi A,Jung S,Man KN,Nair R,Jockusch WJ,Wojcik SM,Brose N,Rhee JS

    更新日期:2012-06-21 00:00:00

  • Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression.

    abstract::Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.35

    authors: Nelson CM,Inman JL,Bissell MJ

    更新日期:2008-01-01 00:00:00

  • Isolation of plasma membrane-associated membranes from rat liver.

    abstract::Dynamic interplay between intracellular organelles requires a particular functional apposition of membrane structures. The organelles involved come into close contact, but do not fuse, thereby giving rise to notable microdomains; these microdomains allow rapid communication between the organelles. Plasma membrane-asso...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2014.016

    authors: Suski JM,Lebiedzinska M,Wojtala A,Duszynski J,Giorgi C,Pinton P,Wieckowski MR

    更新日期:2014-02-01 00:00:00

  • Radiolabeling of HYNIC-annexin V with technetium-99m for in vivo imaging of apoptosis.

    abstract::Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.17

    authors: Blankenberg FG,Vanderheyden JL,Strauss HW,Tait JF

    更新日期:2006-01-01 00:00:00

  • A comprehensive pipeline for translational top-down proteomics from a single blood draw.

    abstract::Top-down proteomics (TDP) by mass spectrometry (MS) is a technique by which intact proteins are analyzed. It has become increasingly popDesalting and concentrating GELFrEEular in translational research because of the value of characterizing distinct proteoforms of intact proteins. Compared to bottom-up proteomics (BUP...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-018-0085-7

    authors: Toby TK,Fornelli L,Srzentić K,DeHart CJ,Levitsky J,Friedewald J,Kelleher NL

    更新日期:2019-01-01 00:00:00

  • Intravascular staining for discrimination of vascular and tissue leukocytes.

    abstract::Characterization of the cellular participants in tissue immune responses is crucial to understanding infection, cancer, autoimmunity, allergy, graft rejection and other immunological processes. Previous reports indicate that leukocytes in lung vasculature fail to be completely removed by perfusion. Several studies sug...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2014.005

    authors: Anderson KG,Mayer-Barber K,Sung H,Beura L,James BR,Taylor JJ,Qunaj L,Griffith TS,Vezys V,Barber DL,Masopust D

    更新日期:2014-01-01 00:00:00