Name: Optische fotothermische infrarood-fluorescentie in situ hybridisatie (OPTIR-FISH) (Video) | JoVE Journal
Uploaded: 2024-02-23T14:20:00.000+00:00
Duration: 4 min 7 s
Description: Leer OPTIR-FISH voor het identificeren van individuele cellen en analyse van metabolisme op enkelcellig niveau. Deze methode biedt inzicht in celinteracties binnen complexe biologische systemen.

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<meta charset="utf8"></head><body>Uitgebreid protocol voor celidentificatie en metabole analyse met behulp van OPTIR-FISH

<ol><li>Evers, T. M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deciphering+metabolic+heterogeneity+by+single-cell+analysis%5BTitle%5D)+AND+%22Anal+Chem%22%5BJournal%5D)">Deciphering metabolic heterogeneity by single-cell analysis.</a> Anal Chem. 91 (21), 13314-13323 (2019).</li>
<li>Overmann, J., Abt, B., Sikorski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Present+and+future+of+culturing+bacteria%5BTitle%5D)+AND+%22Annu+Rev+Microbiol%22%5BJournal%5D)">Present and future of culturing bacteria.</a> Annu Rev Microbiol. 71, 711-730 (2017).</li>
<li>Epstein, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+phenomenon+of+microbial+uncultivability%5BTitle%5D)+AND+%22Curr+Opin+Microbiol%22%5BJournal%5D)">The phenomenon of microbial uncultivability.</a> Curr Opin Microbiol. 16 (5), 636-642 (2013).</li>
<li>Su, C., Lei, L., Duan, Y., Zhang, K. -Q., Yang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Culture-independent+methods+for+studying+environmental+microorganisms%3A+methods%2C+application%2C+and+perspective%5BTitle%5D)+AND+%22Appl+Microbiol+Biotechnol%22%5BJournal%5D)">Culture-independent methods for studying environmental microorganisms: methods, application, and perspective.</a> Appl Microbiol Biotechnol. 93 (3), 993-1003 (2012).</li>
<li>Musat, N., Foster, R., Vagner, T., Adam, B., Kuypers, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Detecting+metabolic+activities+in+single+cells%2C+with+emphasis+on+nanoSIMS%5BTitle%5D)+AND+%22FEMS+Microbiol+Rev%22%5BJournal%5D)">Detecting metabolic activities in single cells, with emphasis on nanoSIMS.</a> FEMS Microbiol Rev. 36 (2), 486-511 (2012).</li>
<li>Jiang, H., Kilburn, M. R., Decelle, J., Musat, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((NanoSIMS+chemical+imaging+combined+with+correlative+microscopy+for+biological+sample+analysis%5BTitle%5D)+AND+%22Curr+Opin+Biotechnol%22%5BJournal%5D)">NanoSIMS chemical imaging combined with correlative microscopy for biological sample analysis.</a> Curr Opin Biotechnol. 41, 130-135 (2016).</li>
<li>Porta Siegel, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mass+spectrometry+imaging+and+integration+with+other+imaging+modalities+for+greater+molecular+understanding+of+biological+tissues%5BTitle%5D)+AND+%22Mol+Imaging+Biol%22%5BJournal%5D)">Mass spectrometry imaging and integration with other imaging modalities for greater molecular understanding of biological tissues.</a> Mol Imaging Biol. 20 (6), 888-901 (2018).</li>
<li>Lagunov, O., Drenchev, N., Chakarova, K., Panayotov, D., Hadjiivanov, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isotopic+labelling+in+vibrational+spectroscopy%3A+a+technique+to+decipher+the+structure+of+surface+species%5BTitle%5D)+AND+%22Top+Catal%22%5BJournal%5D)">Isotopic labelling in vibrational spectroscopy: a technique to decipher the structure of surface species.</a> Top Catal. 60, 1486-1495 (2017).</li>
<li>Bai, Y., Guo, Z., Pereira, F. C., Wagner, M., Cheng, J. -X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mid-infrared+photothermal-fluorescence+in+situ+hybridization+for+functional+analysis+and+genetic+identification+of+single+cells%5BTitle%5D)+AND+%22Anal+Chem%22%5BJournal%5D)">Mid-infrared photothermal-fluorescence in situ hybridization for functional analysis and genetic identification of single cells.</a> Anal Chem. 95 (4), 2398-2405 (2023).</li>
<li>Manz, W., Amann, R., Ludwig, W., Wagner, M., Schleifer, K. -H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Phylogenetic+oligodeoxynucleotide+probes+for+the+major+subclasses+of+proteobacteria%3A+problems+and+solutions%5BTitle%5D)+AND+%22System+Appl+Microbiol%22%5BJournal%5D)">Phylogenetic oligodeoxynucleotide probes for the major subclasses of proteobacteria: problems and solutions.</a> System Appl Microbiol. 15 (4), 593-600 (1992).</li>
<li>Manz, W., Amann, R., Ludwig, W., Vancanneyt, M., Schleifer, K. -H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Application+of+a+suite+of+16S+rRNA-specific+oligonucleotide+probes+designed+to+investigate+bacteria+of+the+phylum+cytophaga-flavobacter-bacteroides+in+the+natural+environment%5BTitle%5D)+AND+%22Microbiology%22%5BJournal%5D)">Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment.</a> Microbiology. 142, 1097-1106 (1996).</li>
<li>Daims, H., Stoecker, K., Wagner, M. Fluorescence In Situ Hybridization for the Detection of Prokaryotes. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aDaims%2C+H+%22Molecular+Microbial+Ecology%22">Molecular Microbial Ecology</a>. , Taylor & Francis. London. (2004).</li>
<li>Bridger, J. M., Volpi, E. V. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aBridger%2C+JM+%22Fluorescence+In+Situ+Hybridization+%28FISH%29%3A+Protocols+and+Applications%22">Fluorescence In Situ Hybridization (FISH): Protocols and Applications</a>. , Springer, Humana. Totowa, NJ. (2010).</li>
<li>Wang, D. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+quick+and+simple+FISH+protocol+with+hybridization-sensitive+fluorescent+linear+oligodeoxynucleotide+probes%5BTitle%5D)+AND+%22RNA%22%5BJournal%5D)">A quick and simple FISH protocol with hybridization-sensitive fluorescent linear oligodeoxynucleotide probes.</a> RNA. 18 (1), 166-175 (2012).</li>
<li>Greuter, D., Loy, A., Horn, M., Rattei, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((probeBase-an+online+resource+for+rRNA-targeted+oligonucleotide+probes+and+primers%3A+new+features+2016%5BTitle%5D)+AND+%22Nucleic+Acids+Res%22%5BJournal%5D)">probeBase-an online resource for rRNA-targeted oligonucleotide probes and primers: new features 2016.</a> Nucleic Acids Res. 44, D586-D589 (2016).</li>
<li>Yilmaz, L. S., Parnerkar, S., Noguera, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((mathFISH%2C+a+web+tool+that+uses+thermodynamics-based+mathematical+models+for+in+silico+evaluation+of+oligonucleotide+probes+for+fluorescence+in+situ+hybridization%5BTitle%5D)+AND+%22Appl+Environ+Microbiol%22%5BJournal%5D)">mathFISH, a web tool that uses thermodynamics-based mathematical models for in silico evaluation of oligonucleotide probes for fluorescence in situ hybridization.</a> Appl Environ Microbiol. 77 (3), 1118-1122 (2011).</li>
<li>Lima, C., Muhamadali, H., Xu, Y., Kansiz, M., Goodacre, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Imaging+isotopically+labeled+bacteria+at+the+single-cell+level+using+high-resolution+optical+infrared+photothermal+spectroscopy%5BTitle%5D)+AND+%22Anal+Chem%22%5BJournal%5D)">Imaging isotopically labeled bacteria at the single-cell level using high-resolution optical infrared photothermal spectroscopy.</a> Anal Chem. 93 (6), 3082-3088 (2021).</li>
<li>Yin, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nanosecond-resolution+photothermal+dynamic+imaging+via+MHZ+digitization+and+match+filtering%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Nanosecond-resolution photothermal dynamic imaging via MHZ digitization and match filtering.</a> Nat Commun. 12 (1), 7097(2021).</li>
<li>Bai, Y., Yin, J., Cheng, J. -X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bond-selective+imaging+by+optically+sensing+the+mid-infrared+photothermal+effect%5BTitle%5D)+AND+%22Sci+Adv%22%5BJournal%5D)">Bond-selective imaging by optically sensing the mid-infrared photothermal effect.</a> Sci Adv. 7 (20), (2021).</li>
<li>Xia, Q., Yin, J., Guo, Z., Cheng, J. -X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mid-infrared+photothermal+microscopy%3A+principle%2C+instrumentation%2C+and+applications%5BTitle%5D)+AND+%22J+Phys+Chemi+B%22%5BJournal%5D)">Mid-infrared photothermal microscopy: principle, instrumentation, and applications.</a> J Phys Chemi B. 126 (43), 8597-8613 (2022).</li>
<li>Du, J., Wang, H., Wei, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bringing+vibrational+imaging+to+chemical+biology+with+molecular+probes%5BTitle%5D)+AND+%22ACS+Chem+Biol%22%5BJournal%5D)">Bringing vibrational imaging to chemical biology with molecular probes.</a> ACS Chem Biol. 17 (7), 1621-1637 (2022).</li>
<li>Miao, Y., Shi, L., Hu, F., Min, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Probe+design+for+super-multiplexed+vibrational+imaging%5BTitle%5D)+AND+%22Phys+Biol%22%5BJournal%5D)">Probe design for super-multiplexed vibrational imaging.</a> Phys Biol. 16 (4), 041003(2019).</li>
<li>Shi, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mid-infrared+metabolic+imaging+with+vibrational+probes%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">Mid-infrared metabolic imaging with vibrational probes.</a> Nat Methods. 17 (8), 844-851 (2020).</li>
<li>Bai, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Single-cell+mapping+of+lipid+metabolites+using+an+infrared+probe+in+human-derived+model+systems%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Single-cell mapping of lipid metabolites using an infrared probe in human-derived model systems.</a> Nat Commun. 15 (1), 350(2024).</li>
<li>Zhang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spectral+tracing+of+deuterium+for+imaging+glucose+metabolism%5BTitle%5D)+AND+%22Nat+Biomed+Eng%22%5BJournal%5D)">Spectral tracing of deuterium for imaging glucose metabolism.</a> Nat Biomed Eng. 3 (5), 402-413 (2019).</li>
<li>Ge, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SRS-FISH%3A+A+high-throughput+platform+linking+microbiome+metabolism+to+identity+at+the+single-cell+level%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">SRS-FISH: A high-throughput platform linking microbiome metabolism to identity at the single-cell level.</a> Proc Natl Acad Sci U S A. 119 (26), e2203519119(2022).</li>
<li>Bai, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ultrafast+chemical+imaging+by+widefield+photothermal+sensing+of+infrared+absorption%5BTitle%5D)+AND+%22Sci+Adv%22%5BJournal%5D)">Ultrafast chemical imaging by widefield photothermal sensing of infrared absorption.</a> Sci Adv. 5 (7), 7127(2019).</li>
<li>Zhang, J., Zhao, J., Lin, H., Tan, Y., Cheng, J. -X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High-speed+chemical+imaging+by+dense-net+learning+of+femtosecond+stimulated+Raman+scattering%5BTitle%5D)+AND+%22J+Phys+Chem+Lett%22%5BJournal%5D)">High-speed chemical imaging by dense-net learning of femtosecond stimulated Raman scattering.</a> J Phys Chem Lett. 11 (20), 8573-8578 (2020).</li>
<li>Lin, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microsecond+fingerprint+stimulated+Raman+spectroscopic+imaging+by+ultrafast+tuning+and+spatial-spectral+learning%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Microsecond fingerprint stimulated Raman spectroscopic imaging by ultrafast tuning and spatial-spectral learning.</a> Nat Commun. 12 (1), 3052(2021).</li>
<li>Fu, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Super-resolution+imaging+of+non-fluorescent+molecules+by+photothermal+relaxation+localization+microscopy%5BTitle%5D)+AND+%22Nat+Photon%22%5BJournal%5D)">Super-resolution imaging of non-fluorescent molecules by photothermal relaxation localization microscopy.</a> Nat Photon. 17, 330-337 (2023).</li>
<li>Tamamitsu, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mid-infrared+widefield+nanoscopy%5BTitle%5D)+AND+%22arXiv%22%5BJournal%5D)">Mid-infrared widefield nanoscopy.</a> arXiv. , (2023).</li>
</ol>

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optische fotothermische infrarood-fluorescentie in situ hybridisatie (optir-fish)

Optical Photothermal Infrared-Fluorescence In Situ Hybridization (OPTIR-FISH)

Optische fotothermische infrarood-fluorescentie in situ hybridisatie (OPTIR-FISH)

Understanding the metabolic activities of individual cells within complex communities is critical for unraveling their role in human disease. Here, we ...

optical-photothermal-infrared-fluorescence-situ-hybridization-optir

Research

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Bioengineering

Bio-engineering

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Dubbele Fluorescentie In situ Hybridisatie in Fresh Brain secties

Detectie van virale RNA door fluorescentie<em&gt; In situ</em&gt; Hybridisatie (FISH)

Hier beschrijven we een geheel gemonteerde...

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Fluorescentie<em&gt; In situ</em&gt; Hybridisaties (FISH) voor de lokalisatie van Virussen en Endosymbiotic Bacteriën in Plant en Insect Tissues

Het beschreven RNA in situ hybridisatie protocol staat toe de opsporing van RNA in hele Drosophila embryo's of ontleed weefsels. Met behulp van 96-wells microtiterplaat platen en tyramide signaal versterking, kunnen afschriften worden gedetecteerd op hoge resolutie, gevoeligheid en doorvoer, en tegen een relatief lage prijs.

Hoge resolutie fluorescentie In Situ hybridisatie in Drosophila embryo's en weefsels met behulp van Tyramide signaal versterking

Dit manuscript presenteert een gedetailleerde methode voor het genereren van X-chromosoom arm sondes en presterende fluorescentie in situ hybridisatie (FISH) te onderzoeken van de Braziliaanse deelstaat zus chromatide cohesie in prometaphase en metafase ik gearresteerd  Drosophila eicellen. Dit protocol is geschikt om te bepalen of cohesie Meiotische arm intact of verstoord in verschillende genotypen is.

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Expansie-ondersteunde iteratieve fluorescentie in situ hybridisatie (EASI-FISH) is een robuuste techniek die expansiemicroscopie combineert met fluorescentie in situ hybridisatie (FISH) om de expressie van meerdere genen in dik weefsel te detecteren. Dit protocol schetst recente vorderingen in de methode en de aanpassing ervan voor het labelen van het intacte Drosophila centrale zenuwstelsel..

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Optische resolutie fotoakoestische microscopie (OR-PAM) is een opkomende technologie die in staat van beeldvorming optische absorptie...

Drie-dimensionale Optische resolutie Fotoakoestisch Microscopie

Leer OPTIR-FISH voor het identificeren van individuele cellen en analyse van metabolisme op enkelcellig niveau. Deze methode biedt inzicht in celinteracties binnen complexe biologische systemen.

Optische fotothermische infrarood-fluorescentie in situ hybridisatie (OPTIR-FISH)

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