Name: Infant Immuniteit in kaart brengen met minimale input: Integratieve single-cell en multiomic profiling (Video) | JoVE Journal
Uploaded: 2026-04-03T12:35:00.000+00:00
Duration: 10 min 29 s
Description: Leer hoe je hoog-dimensionale immuunprofilering uitvoert bij pasgeborenen met minimale bloedvolumes via single-cell en multiomic technieken. Deze methode ondersteunt longitudinaal onderzoek naar vroege immuunontwikkeling.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

<ol><li>Kollmann, T. R., Kampmann, B., Mazmanian, S. K., Marchant, A., Levy, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protecting+the+newborn+and+young+infant+from+infectious+diseases%3A+lessons+from+immune+ontogeny%5BTitle%5D)+AND+%22Immunity%22%5BJournal%5D)">Protecting the newborn and young infant from infectious diseases: lessons from immune ontogeny.</a> Immunity. 46 (3), 350-363 (2017).</li>
<li>Jain, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+early-life+education+of+the+immune+system%3A+moms%2C+microbes+and+%28missed%29+opportunities%5BTitle%5D)+AND+%22Gut+Microbes%22%5BJournal%5D)">The early-life education of the immune system: moms, microbes and (missed) opportunities.</a> Gut Microbes. 12 (1), 1824564(2020).</li>
<li>Konnikova, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fetal+and+neonatal+T+cell+immunity%3A+unique+features+and+challenges%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">Fetal and neonatal T cell immunity: unique features and challenges.</a> Front Immunol. 11, 594939(2020).</li>
<li>Pieren, D. K. J., Boer, M. C., De Wit, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+adaptive+immune+system+in+early+life%3A+the+shift+makes+it+count%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">The adaptive immune system in early life: the shift makes it count.</a> Front Immunol. 13, 1031924(2022).</li>
<li>Brodsky, N. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Early-life+human+CD8%2B+T+cells+exhibit+rapid%2C+short-lived+effector+responses+and+a+unique+transcription+factor+landscape%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Early-life human CD8+ T cells exhibit rapid, short-lived effector responses and a unique transcription factor landscape.</a> Proc Natl Acad Sci U S A. 122 (31), e2421106122(2025).</li>
<li>Olaloye, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+single-cell+atlas+of+circulating+immune+cells+over+the+first+2+months+of+age+in+extremely+premature+infants%5BTitle%5D)+AND+%22Sci+Transl+Med%22%5BJournal%5D)">A single-cell atlas of circulating immune cells over the first 2 months of age in extremely premature infants.</a> Sci Transl Med. 17, eadr0942(2025).</li>
<li>Watson, N. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+gene+regulatory+basis+of+bystander+activation+in+CD8%2B+T+cells%5BTitle%5D)+AND+%22Sci+Immunol%22%5BJournal%5D)">The gene regulatory basis of bystander activation in CD8+ T cells.</a> Sci Immunol. 9 (92), eadf8776(2024).</li>
<li>Maymí, V. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neonatal+CD8%2B+T+cells+resist+exhaustion+during+chronic+infection%5BTitle%5D)+AND+%22J+Immunol%22%5BJournal%5D)">Neonatal CD8+ T cells resist exhaustion during chronic infection.</a> J Immunol. 212 (5), 834-843 (2024).</li>
<li>Kandasamy, R., Kollmann, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Challenges+in+neonatal+immunology%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">Challenges in neonatal immunology.</a> Front Immunol. 8, 1756(2017).</li>
<li>Howie, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Blood+sample+volumes+in+child+health+research%3A+review+of+safe+limits%5BTitle%5D)+AND+%22Bull+World+Health+Organ%22%5BJournal%5D)">Blood sample volumes in child health research: review of safe limits.</a> Bull World Health Organ. 89 (1), 46-53 (2011).</li>
<li>Gross, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Researching+COVID+to+enhance+recovery+%28RECOVER%29+pediatric+study+protocol%3A+rationale%2C+objectives+and+design%5BTitle%5D)+AND+%22medRxiv%22%5BJournal%5D)">Researching COVID to enhance recovery (RECOVER) pediatric study protocol: rationale, objectives and design.</a> medRxiv. , (2023).</li>
<li>Bourguignon, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Processing+of+blood+samples+influences+PBMC+viability+and+outcome+of+cell-mediated+immune+responses+in+antiretroviral+therapy-na%C3%AFve+HIV-1-infected+patients%5BTitle%5D)+AND+%22J+Immunol+Methods%22%5BJournal%5D)">Processing of blood samples influences PBMC viability and outcome of cell-mediated immune responses in antiretroviral therapy-naïve HIV-1-infected patients.</a> J Immunol Methods. 414, 1-10 (2014).</li>
<li>Urbansky, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rapid+and+effective+enrichment+of+mononuclear+cells+from+blood+using+acoustophoresis%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Rapid and effective enrichment of mononuclear cells from blood using acoustophoresis.</a> Sci Rep. 7 (1), 17161(2017).</li>
<li>Najar, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Proliferative+and+phenotypical+characteristics+of+human+adipose+tissue-derived+stem+cells%3A+comparison+of+purification+methods%5BTitle%5D)+AND+%22Cytotherapy%22%5BJournal%5D)">Proliferative and phenotypical characteristics of human adipose tissue-derived stem cells: comparison of purification methods.</a> Cytotherapy. 16 (9), 1220-1228 (2014).</li>
<li>Abebe, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Diagnostic+significance+of+complete+blood+cell+count+and+hemogram-derived+markers+for+neonatal+sepsis%5BTitle%5D)+AND+%22World+J+Clin+Pediatr%22%5BJournal%5D)">Diagnostic significance of complete blood cell count and hemogram-derived markers for neonatal sepsis.</a> World J Clin Pediatr. 13 (2), 1-13 (2024).</li>
<li>Martino, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DNA+methylation+signatures+underpinning+blood+neutrophil+to+lymphocyte+ratio+during+first+week+of+human+life%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">DNA methylation signatures underpinning blood neutrophil to lymphocyte ratio during first week of human life.</a> Nat Commun. 15, 8157(2024).</li>
<li>Jones, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DNA+methylation+differences+between+cord+and+adult+white+blood+cells+reflect+postnatal+immune+cell+maturation%5BTitle%5D)+AND+%22Commun+Biol%22%5BJournal%5D)">DNA methylation differences between cord and adult white blood cells reflect postnatal immune cell maturation.</a> Commun Biol. 8, 237(2025).</li>
<li>Lee, A. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dynamic+molecular+changes+during+the+first+week+of+human+life+follow+a+robust+developmental+trajectory%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Dynamic molecular changes during the first week of human life follow a robust developmental trajectory.</a> Nat Commun. 10, 1092(2019).</li>
<li>Mithal, L. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evolution+of+the+umbilical+cord+blood+proteome+across+gestational+development%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Evolution of the umbilical cord blood proteome across gestational development.</a> Sci Rep. 15 (1), 2233(2025).</li>
<li>Olin, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Stereotypic+immune+system+development+in+newborn+children%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Stereotypic immune system development in newborn children.</a> Cell. 174 (5), 1277-1292.e14 (2018).</li>
<li>Spasic, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spectral+flow+cytometry+methods+and+pipelines+for+comprehensive+immunoprofiling%5BTitle%5D)+AND+%22Cancer+Res+Commun%22%5BJournal%5D)">Spectral flow cytometry methods and pipelines for comprehensive immunoprofiling.</a> Cancer Res Commun. 4 (3), 895-910 (2024).</li>
<li>Park, L. M., Lannigan, J., Jaimes, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((OMIP-069%3A+forty-color+full+spectrum+flow+cytometry+panel%5BTitle%5D)+AND+%22Cytometry+A%22%5BJournal%5D)">OMIP-069: forty-color full spectrum flow cytometry panel.</a> Cytometry A. 97 (10), 1044-1051 (2020).</li>
<li>Sahir, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Development+of+a+43-color+panel+for+comprehensive+immune+cell+characterization%5BTitle%5D)+AND+%22Cytometry+A%22%5BJournal%5D)">Development of a 43-color panel for comprehensive immune cell characterization.</a> Cytometry A. , (2020).</li>
<li>Tang, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((mRNA-seq+whole-transcriptome+analysis+of+a+single+cell%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">mRNA-seq whole-transcriptome analysis of a single cell.</a> Nat Methods. 6, 377-382 (2009).</li>
<li>Wolf, F. A., Angerer, P., Theis, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SCANPY%3A+large-scale+single-cell+gene+expression+data+analysis%5BTitle%5D)+AND+%22Genome+Biol%22%5BJournal%5D)">SCANPY: large-scale single-cell gene expression data analysis.</a> Genome Biol. 19, 15(2018).</li>
<li>Wolock, S. L., Lopez, R., Klein, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Scrublet%3A+computational+identification+of+cell+doublets%5BTitle%5D)+AND+%22Cell+Syst%22%5BJournal%5D)">Scrublet: computational identification of cell doublets.</a> Cell Syst. 8 (4), 281-291.e9 (2019).</li>
<li>Polanski, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((BBKNN%3A+fast+batch+alignment+of+single-cell+transcriptomes%5BTitle%5D)+AND+%22Bioinformatics%22%5BJournal%5D)">BBKNN: fast batch alignment of single-cell transcriptomes.</a> Bioinformatics. 36 (3), 964-965 (2020).</li>
<li>Li, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DMSO-free+cryopreservation+improves+recovery+and+function+of+human+T+and+NK+cells%5BTitle%5D)+AND+%22Cell+Transplant%22%5BJournal%5D)">DMSO-free cryopreservation improves recovery and function of human T and NK cells.</a> Cell Transplant. 31, 1-12 (2022).</li>
<li>Meryman, H. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cryopreservation+of+living+cells%3A+principles+and+practice%5BTitle%5D)+AND+%22Transfusion%22%5BJournal%5D)">Cryopreservation of living cells: principles and practice.</a> Transfusion. 47 (5), 935-945 (2007).</li>
<li>Wohnhaas, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((RNA+integrity+in+cryopreserved+immune+cells+enables+robust+single-cell+transcriptomics%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">RNA integrity in cryopreserved immune cells enables robust single-cell transcriptomics.</a> Front Immunol. 10, 204(2019).</li>
<li>Daae, L. N. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comparison+of+haematological+parameters+in+capillary+and+venous+blood+samples+from+hospitalized+children%5BTitle%5D)+AND+%22Scand+J+Clin+Lab+Invest%22%5BJournal%5D)">Comparison of haematological parameters in capillary and venous blood samples from hospitalized children.</a> Scand J Clin Lab Invest. 51 (7), 651-654 (1991).</li>
<li>Meites, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skin-puncture+and+blood-collecting+technique+for+infants%5BTitle%5D)+AND+%22Clin+Chem%22%5BJournal%5D)">Skin-puncture and blood-collecting technique for infants.</a> Clin Chem. 34 (9), 1890-1894 (1988).</li>
<li>Grüner, N., Stambouli, O., Ross, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dried+blood+spots%3A+preparing+and+processing%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Dried blood spots: preparing and processing.</a> J Vis Exp. (97), e52619(2015).</li>
<li>Waaijer, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spectral+flow+cytometry+panel+for+comprehensive+immunophenotyping%5BTitle%5D)+AND+%22Cytometry+A%22%5BJournal%5D)">Spectral flow cytometry panel for comprehensive immunophenotyping.</a> Cytometry A. 107 (4), 226-232 (2025).</li>
<li>Thieme-Ehlert, M. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((OMIP-114%3A+a+36-color+spectral+flow+cytometry+panel%5BTitle%5D)+AND+%22Cytometry+A%22%5BJournal%5D)">OMIP-114: a 36-color spectral flow cytometry panel.</a> Cytometry A. 107 (6), 364-371 (2025).</li>
<li>Yamawaki, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Systematic+comparison+of+high-throughput+single-cell+RNA-seq+methods%5BTitle%5D)+AND+%22BMC+Genomics%22%5BJournal%5D)">Systematic comparison of high-throughput single-cell RNA-seq methods.</a> BMC Genomics. 22, 66(2021).</li>
<li>Stoeckius, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Simultaneous+epitope+and+transcriptome+measurement+in+single+cells%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">Simultaneous epitope and transcriptome measurement in single cells.</a> Nat Methods. 14 (9), 865-868 (2017).</li>
<li>Grandi, F. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chromatin+accessibility+profiling+by+ATAC-seq%5BTitle%5D)+AND+%22Nat+Protoc%22%5BJournal%5D)">Chromatin accessibility profiling by ATAC-seq.</a> Nat Protoc. 17 (6), 1518-1552 (2022).</li>
<li>Kim, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Perspectives+on+single-nucleus+RNA+sequencing%5BTitle%5D)+AND+%22J+Pathol+Transl+Med%22%5BJournal%5D)">Perspectives on single-nucleus RNA sequencing.</a> J Pathol Transl Med. 57 (1), 52-59 (2023).</li>
</ol>

<table><tbody><tr><td>0.5 M EDTA, pH 8.0</td><td>Thermo Fisher Scientific&nbsp;</td><td>15575-038&nbsp;</td><td></td></tr><tr><td>1x RBC Lysis Buffer&nbsp;</td><td>Invitrogen&nbsp;</td><td>3177165&nbsp;</td><td></td></tr><tr><td>BCA Protein Assay Kit &nbsp;</td><td>Abcam&nbsp;</td><td>ab287853&nbsp;</td><td></td></tr><tr><td>BD Microtainer MAP Microtube</td><td>Fisher Scientific&nbsp;</td><td>22-253-270</td><td></td></tr><tr><td>Bel-Art Cryo-Safe -1&deg;C Freeze Controller&nbsp;</td><td>Millipore Sigma&nbsp;</td><td>41122800&nbsp;</td><td></td></tr><tr><td>Bovine Serum Albumin (BSA)</td><td>Sigma-Aldrich&nbsp;</td><td>A9647-100G&nbsp;</td><td></td></tr><tr><td>Brilliant Ultra Violet 496 Anti-Human CD3&nbsp;</td><td>Thermo Fisher Scientific&nbsp;</td><td>364-0038-42&nbsp;</td><td></td></tr><tr><td>Brilliant Ultra Violet 661 anti-Human CD11c&nbsp;</td><td>BD BioScience&nbsp;</td><td>612968&nbsp;</td><td></td></tr><tr><td>Brilliant Vil750 Anti-Human CD14&nbsp;</td><td>BioLegend&nbsp;</td><td>367135&nbsp;</td><td></td></tr><tr><td>Brilliant Violet 570 Anti-Human CD8a&nbsp;</td><td>BioLegend&nbsp;</td><td>301038&nbsp;</td><td></td></tr><tr><td>Brilliant Violet 711 Anti-Human CD56&nbsp;</td><td>BioLegend&nbsp;</td><td>318336&nbsp;</td><td></td></tr><tr><td>cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail</td><td>Millipore Sigma&nbsp;</td><td>11836170001&nbsp;</td><td></td></tr><tr><td>Dead Cell Removal Kit&nbsp;</td><td>Miltenyi Biotec&nbsp;</td><td>130-090-101&nbsp;</td><td></td></tr><tr><td>Dimethyl Sulfoxide (DMSO)&nbsp;</td><td>Thermo Fisher Scientific&nbsp;</td><td>BP231-100&nbsp;</td><td></td></tr><tr><td>Dulbecco&rsquo;s Phosphate buffered saline (DPBS), no calcium, no magnesium&nbsp;</td><td>Thermo Fisher Scientific&nbsp;</td><td>14190144&nbsp;</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS)&nbsp;</td><td>Gibco&nbsp;</td><td>A5256801&nbsp;</td><td></td></tr><tr><td>Fisherbrand Digital Vortex Mixer&nbsp;</td><td>Thermo Fisher Scientific&nbsp;</td><td>02215418&nbsp;</td><td></td></tr><tr><td>Fixation Buffer&nbsp;</td><td>BioLegend&nbsp;</td><td>420801&nbsp;</td><td></td></tr><tr><td>GlutaMAX Supplement</td><td>Gibco&nbsp;</td><td>35050061&nbsp;</td><td></td></tr><tr><td>MACS MultiStand&nbsp;</td><td>Miltenyi Biotec&nbsp;</td><td>130-042-303&nbsp;</td><td></td></tr><tr><td>MS columns&nbsp;</td><td>Miltenyi Biotec&nbsp;</td><td>130-042-201&nbsp;</td><td></td></tr><tr><td>Name</td><td>Company</td><td>Catalog Number</td><td></td></tr><tr><td>OctoMACS Separator&nbsp;</td><td>Miltenyi Biotec&nbsp;</td><td>130-042-109&nbsp;</td><td></td></tr><tr><td>Pacific Blue Anti-Human CD4&nbsp;</td><td>BioLegend&nbsp;</td><td>344619&nbsp;</td><td></td></tr><tr><td>Penicillin-Streptomycin (10,000 U/mL)</td><td>Thermo Fisher Scientific&nbsp;</td><td>15140-122&nbsp;</td><td></td></tr><tr><td>RB780 Anti-Human CD33&nbsp;</td><td>BD Biosciences&nbsp;</td><td>755606&nbsp;</td><td></td></tr><tr><td>Rierdge 3mm Metal Puncher&nbsp;</td><td>Rierdge&nbsp;</td><td>sdd231031pc-01&nbsp;</td><td></td></tr><tr><td>RPMI 1640 Medium</td><td>Gibco&nbsp;</td><td>11875-093&nbsp;</td><td></td></tr><tr><td>Spark UV 387 Anti-Human CD19&nbsp;</td><td>BioLegend&nbsp;</td><td>302289&nbsp;</td><td></td></tr><tr><td>Tasso Collection Kits</td><td>Tasso&nbsp;</td><td>N/A</td><td></td></tr><tr><td>Thermo Scientific Screw Cap Micro Tubes</td><td>Thermo Fisher Scientific&nbsp;</td><td>14-755-287&nbsp;</td><td></td></tr><tr><td>Whatman 309 Proteinsaver</td><td>Cytiva&nbsp;</td><td>41111700&nbsp;</td><td></td></tr><tr><td>Zombie NIR Fixable Viability Kit&nbsp;</td><td>BioLegend&nbsp;</td><td>423105&nbsp;</td><td></td></tr><tr><td>Software en pakketten</td><td></td><td></td><td></td></tr><tr><td>Adobe Illustrator</td><td>2024</td><td></td><td></td></tr><tr><td>bbknn</td><td>1.5.1</td><td></td><td></td></tr><tr><td>FlowJo</td><td>10.8.1</td><td></td><td></td></tr><tr><td>GraphPad Prism</td><td>9.4.0</td><td></td><td></td></tr><tr><td>Name</td><td>Version</td><td></td><td></td></tr><tr><td>Scanpy</td><td>1.9.1</td><td></td><td></td></tr><tr><td>Srublet</td><td>0.2.3</td><td></td><td></td></tr><tr><td>Commerciële assays</td><td></td><td></td><td></td></tr><tr><td>Chromium Single Cell 5&rsquo; Kit</td><td>10X Genomics</td><td>PN-1000263</td><td></td></tr><tr><td>Olink&nbsp;Reveal or Target 96 Inflammatory panel</td><td>Olink</td><td>https://olink.com/products/olink-target-96</td><td></td></tr></tbody></table>

infant immuniteit in kaart brengen met minimale input: integratieve single-cell en multiomic profiling

Mapping Infant Immunity with Minimal Input: Integrative Single-Cell and Multiomic Profiling

Infant Immuniteit in kaart brengen met minimale input: Integratieve single-cell en multiomic profiling

At birth, the neonatal immune system is abruptly confronted with a radically different environment, rich in microbial and environmental stimuli. ...

mapping-infant-immunity-with-minimal-input-integrative-single-cell

Research

JoVE Journal

Immunology and Infection

Immunologie en Infectie

Dit protocol beschrijft hoe de expressie van een groot scala van genen op het klonale niveau te beoordelen. Single-cell RT-qPCR levert zeer betrouwbare resultaten met een sterke gevoeligheid voor honderden monsters en genen.

Eencellige genexpressie met behulp van Multiplex RT-qPCR heterogeniteit van Rare lymfoïde populaties karakteriseren

Hier beschrijven we een eencellige proteomic aanpak om te evalueren van immuun fenotypische en functionele (intracellulaire cytokine inductie) veranderingen in het perifere bloedmonsters geanalyseerd via massale cytometry.

Eencellige analyse van Immunophenotype en Cytokine productie in perifere bloed via massale Cytometry

Bulk gen expressie metingen wolk individuele cel verschillen in heterogene cel populaties. Hier beschrijven we een protocol voor hoe eencellige gen expressie analyse en index Sorteren door bloeien geactiveerd Cell Sorting (FACS) kan worden gecombineerd om te bakenen heterogeniteit en immunophenotypically karakteriseren moleculair afzonderlijke cel populaties.

Een combinatorische eencellige aanpak te karakteriseren de moleculaire en Immunophenotypic heterogeniteit van menselijke stamcellen en voorlopercellen populaties

Kwantitatieve multiplex Immunoprecipitation (QMI) gebruikt Flowcytometrie voor gevoelige detectie van verschillen in de overvloed van gerichte eiwit-eiwit interacties tussen twee monsters. QMI kan worden uitgevoerd met een kleine hoeveelheid biomateriaal, vereist geen genetisch gemanipuleerde Tags en kan worden aangepast voor elk eerder gedefinieerd proteïne-interactie netwerk.

Kwantificering van eiwit interactie netwerk dynamiek met Multiplexed co-immunoprecipitatie

Het gebruik van een RNA-gebaseerde benadering om kwantitatieve immuunprofielen van vaste tumorweefsels te bepalen en klinische cohorten te benutten voor immuun-oncologische biomarkerontdekking wordt beschreven door middel van een moleculaire en informaticaprotocollen.

Voorspellende immuunmodellering van vaste tumoren

Hier beschrijven we een eenvoudige en toegankelijke strategie voor het visualiseren, kwantificeren en in kaart brengen van immuuncellen in formaline-vaste paraffine-ingebedde tumorweefselsecties. Deze methodologie combineert bestaande beeldvormings- en digitale analysetechnieken met als doel de multiplexingcapaciteit en multiparameteranalyse van beeldvormingstesten uit te breiden.

Visualisatie, kwantificering en mapping van immuuncelpopulaties in de tumormicro-omgeving

In het tijdperk van immunotherapie en eencellige genomische profilering vereist kankerbiologie nieuwe in vitro en computationele hulpmiddelen voor het onderzoeken van de tumor-immuuninterface in een juiste spatiotemporale context. We beschrijven protocollen om tumor-immuun microfluïdische co-culturen te exploiteren in 2D- en 3D-instellingen, compatibel met dynamische, multiparametrische monitoring van cellulaire functies.

Microfluïdische co-cultuurmodellen voor het ontleden van de immuunrespons in in vitro tumormicro-omgevingen

Dit protocol combineert flowcytometrie en eencellige RNA-sequencing om microgliale celtoestanden in het cerebellum van vroege postnatale muizenhersenen te isoleren en te karakteriseren. Het maakt gebruik van enzymatische dissociatie, Percoll-centrifugatie en immunokleuring om de heterogeniteit van microglia's te onthullen en het begrip van hun rol in de ontwikkeling en ziekte van cerebellaire patiënten te verbeteren.

Flowcytometrie en eencellige analyse voor het karakteriseren van microglia-activering bij vroege postnatale hersenontwikkeling bij muizen

Een cel dood-gebaseerde test voor het PTI in Nicotiana benthamiana planten is beschreven.

Test voor Pathogeen-geassocieerde Moleculaire Pattern (PAMP)-Triggered Immuniteit (PTI) in Planten

Selectie van Plasmodium falciparum Parasieten voor Cytoadhesion Human Brain endotheelcellen

Leer hoe je hoog-dimensionale immuunprofilering uitvoert bij pasgeborenen met minimale bloedvolumes via single-cell en multiomic technieken. Deze methode ondersteunt longitudinaal onderzoek naar vroege immuunontwikkeling.

Infant Immuniteit in kaart brengen met minimale input: Integratieve single-cell en multiomic profiling

Meer video's verkennen