Name: Absolute DNA-dichtheid in celkernen in kaart brengen met behulp van lokalisatiemicroscopie met één molecuul (Video) | JoVE Journal
Uploaded: 2025-11-11T14:00:00.000+00:00
Duration: 10 min 57 s
Description: Leer absolute DNA-dichtheid meten in celkernen met behulp van Voronoi-tessellatie en single-molecule localisatiemicroscopie. Deze methode biedt inzicht in chromatine-architectuur en ruimtelijke beperkingen.

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>Passarge, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Emil+Heitz+and+the+concept+of+heterochromatin%3A+longitudinal+chromosome+differentiation+was+recognized+fifty+years+ago%5BTitle%5D)+AND+%22American+Journal+of+Human+Genetics%22%5BJournal%5D)">Emil Heitz and the concept of heterochromatin: longitudinal chromosome differentiation was recognized fifty years ago.</a> American Journal of Human Genetics. 31 (2), 106-115 (1979).</li>
<li>Cremer, T., Cremer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chromosome+territories%5BTitle%5D)+AND+%22Cold+Spring+Harbor+Perspectives+in+Biology%22%5BJournal%5D)">Chromosome territories.</a> Cold Spring Harbor Perspectives in Biology. 2 (3), 003889(2010).</li>
<li>Vecchio, L., Solimando, L., Biggiogera, M., Fakan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Use+of+halogenated+precursors+for+simultaneous+DNA+and+RNA+detection+by+means+of+immunoelectron+and+immunofluorescence+microscopy%5BTitle%5D)+AND+%22The+Journal+of+Histochemistry+and+Cytochemistry%22%5BJournal%5D)">Use of halogenated precursors for simultaneous DNA and RNA detection by means of immunoelectron and immunofluorescence microscopy.</a> The Journal of Histochemistry and Cytochemistry. 56 (1), 45-55 (2008).</li>
<li>Solimando, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spatial+organization+of+nucleotide+excision+repair+proteins+after+UV-induced+DNA+damage+in+the+human+cell+nucleus%5BTitle%5D)+AND+%22Journal+of+Cell+Science%22%5BJournal%5D)">Spatial organization of nucleotide excision repair proteins after UV-induced DNA damage in the human cell nucleus.</a> Journal of Cell Science. 122, 83-91 (2009).</li>
<li>Niedojadlo, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Transcribed+DNA+is+preferentially+located+in+the+perichromatin+region+of+mammalian+cell+nuclei%5BTitle%5D)+AND+%22Experimental+Cell+Research%22%5BJournal%5D)">Transcribed DNA is preferentially located in the perichromatin region of mammalian cell nuclei.</a> Experimental Cell Research. 317 (4), 433-444 (2011).</li>
<li>Padeken, J., Heun, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nucleolus+and+nuclear+periphery%3A+velcro+for+heterochromatin%5BTitle%5D)+AND+%22Current+Opinion+in+Cell+Biology%22%5BJournal%5D)">Nucleolus and nuclear periphery: velcro for heterochromatin.</a> Current Opinion in Cell Biology. 28, 54-60 (2014).</li>
<li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Overview+of+histone+modification%5BTitle%5D)+AND+%22Advances+in+Experimental+Medicine+and+Biology%22%5BJournal%5D)">Overview of histone modification.</a> Advances in Experimental Medicine and Biology. 1283, 1-16 (2021).</li>
<li>Ilango, S., Paital, B., Jayachandran, P., Padma, P. R., Nirmaladevi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epigenetic+alterations+in+cancer%5BTitle%5D)+AND+%22Frontiers+in+Bioscience+%28Landmark+Ed%29%22%5BJournal%5D)">Epigenetic alterations in cancer.</a> Frontiers in Bioscience (Landmark Ed). 25 (6), 1058-1109 (2020).</li>
<li>Saul, D., Kosinsky, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epigenetics+of+aging+and+aging-associated+diseases%29%5BTitle%5D)+AND+%22International+Journal+of+Molecular+Sciences%22%5BJournal%5D)">Epigenetics of aging and aging-associated diseases).</a> International Journal of Molecular Sciences. 22 (1), 401(2021).</li>
<li>Eckersley-Maslin, M. A., Alda-Catalinas, C., Reik, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dynamics+of+the+epigenetic+landscape+during+the+maternal-to-zygotic+transition%5BTitle%5D)+AND+%22Nature+Reviews%2C+Molecular+Cell+Biology%22%5BJournal%5D)">Dynamics of the epigenetic landscape during the maternal-to-zygotic transition.</a> Nature Reviews, Molecular Cell Biology. 19 (7), 436-450 (2018).</li>
<li>Stevens, T. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((3D+structures+of+individual+mammalian+genomes+studied+by+single-cell+Hi-C%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">3D structures of individual mammalian genomes studied by single-cell Hi-C.</a> Nature. 544 (7648), 59-64 (2017).</li>
<li>Lieberman-Aiden, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comprehensive+mapping+of+long-range+interactions+reveals+folding+principles+of+the+human+genome%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Comprehensive mapping of long-range interactions reveals folding principles of the human genome.</a> Science. 326 (5950), 289-293 (2009).</li>
<li>Cremer, C., Masters, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Resolution+enhancement+techniques+in+microscopy%5BTitle%5D)+AND+%22The+European+Physical+Journal+H%22%5BJournal%5D)">Resolution enhancement techniques in microscopy.</a> The European Physical Journal H. 38 (3), 281-344 (2013).</li>
<li>Lelek, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Single-molecule+localization+microscopy%5BTitle%5D)+AND+%22Nature+Reviews+Methods+Primers%22%5BJournal%5D)">Single-molecule localization microscopy.</a> Nature Reviews Methods Primers. 1 (1), 39(2021).</li>
<li>Huang, B., Wang, W., Bates, M., Zhuang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Three-dimensional+super-resolution+imaging+by+stochastic+optical+reconstruction+microscopy%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy.</a> Science. 319 (5864), 810-813 (2008).</li>
<li>Szczurek, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Super-resolution+binding+activated+localization+microscopy+through+reversible+change+of+DNA+conformation%5BTitle%5D)+AND+%22Nucleus%22%5BJournal%5D)">Super-resolution binding activated localization microscopy through reversible change of DNA conformation.</a> Nucleus. 9 (1), 182-189 (2018).</li>
<li>Szczurek, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Imaging+chromatin+nanostructure+with+binding-activated+localization+microscopy+based+on+DNA+structure+fluctuations%5BTitle%5D)+AND+%22Nucleic+Acids+Research%22%5BJournal%5D)">Imaging chromatin nanostructure with binding-activated localization microscopy based on DNA structure fluctuations.</a> Nucleic Acids Research. 45 (8), 56(2017).</li>
<li>Levet, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SR-Tesseler%3A+a+method+to+segment+and+quantify+localization-based+super-resolution+microscopy+data%5BTitle%5D)+AND+%22Nature+Methods%22%5BJournal%5D)">SR-Tesseler: a method to segment and quantify localization-based super-resolution microscopy data.</a> Nature Methods. 12 (11), 1065-1071 (2015).</li>
<li>Roukos, V., Pegoraro, G., Voss, T. C., Misteli, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cell+cycle+staging+of+individual+cells+by+fluorescence+microscopy%5BTitle%5D)+AND+%22Nature+Protocols%22%5BJournal%5D)">Cell cycle staging of individual cells by fluorescence microscopy.</a> Nature Protocols. 10 (2), 334-348 (2015).</li>
<li>Stirling, D. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CellProfiler+4%3A+improvements+in+speed%2C+utility+and+usability%5BTitle%5D)+AND+%22BMC+Bioinformatics%22%5BJournal%5D)">CellProfiler 4: improvements in speed, utility and usability.</a> BMC Bioinformatics. 22 (1), 433(2021).</li>
<li>Team, R. C. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aTeam%2C+RC+%22R%3A+A+Language+and+Environment+for+Statistical+Computing%22">R: A Language and Environment for Statistical Computing</a>. , (2016).</li>
<li>Thompson, R. E., Larson, D. R., Webb, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Precise+nanometer+localization+analysis+for+individual+fluorescent+probes%5BTitle%5D)+AND+%22Biophysical+Journal%22%5BJournal%5D)">Precise nanometer localization analysis for individual fluorescent probes.</a> Biophysical Journal. 82 (5), 2775-2783 (2002).</li>
<li>Schneider, C. A., Rasband, W. S., Eliceiri, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((NIH+Image+to+ImageJ%3A+25+years+of+image+analysis%5BTitle%5D)+AND+%22Nature+Methods%22%5BJournal%5D)">NIH Image to ImageJ: 25 years of image analysis.</a> Nature Methods. 9 (7), 671-675 (2012).</li>
<li>Ovesny, M., Krizek, P., Borkovec, J., Svindrych, Z., Hagen, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ThunderSTORM%3A+a+comprehensive+ImageJ+plug-in+for+PALM+and+STORM+data+analysis+and+super-resolution+imaging%5BTitle%5D)+AND+%22Bioinformatics%22%5BJournal%5D)">ThunderSTORM: a comprehensive ImageJ plug-in for PALM and STORM data analysis and super-resolution imaging.</a> Bioinformatics. 30 (16), 2389-2390 (2014).</li>
<li>Schindelin, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fiji%3A+an+open-source+platform+for+biological-image+analysis%5BTitle%5D)+AND+%22Nature+Methods%22%5BJournal%5D)">Fiji: an open-source platform for biological-image analysis.</a> Nature Methods. 9 (7), 676-682 (2012).</li>
<li>Maeshima, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+physical+size+of+transcription+factors+is+key+to+transcriptional+regulation+in+chromatin+domains%5BTitle%5D)+AND+%22Journal+of+Physics.+Condensed+Matter%22%5BJournal%5D)">The physical size of transcription factors is key to transcriptional regulation in chromatin domains.</a> Journal of Physics. Condensed Matter. 27 (6), 064116(2015).</li>
<li>Itoh, Y., Woods, E. J., Minami, K., Maeshima, K., Collepardo-Guevara, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Liquid-like+chromatin+in+the+cell%3A+What+can+we+learn+from+imaging+and+computational+modeling%5BTitle%5D)+AND+%22Current+Opinion+in+Structural+Biology%22%5BJournal%5D)">Liquid-like chromatin in the cell: What can we learn from imaging and computational modeling.</a> Current Opinion in Structural Biology. 71, 123-135 (2021).</li>
<li>Strickfaden, H., Xu, Z. Z., Hendzel, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Visualization+of+miniSOG+tagged+DNA+repair+proteins+in+combination+with+Electron+Spectroscopic+Imaging+%28ESI%29%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Visualization of miniSOG tagged DNA repair proteins in combination with Electron Spectroscopic Imaging (ESI).</a> Journal of Visualized Experiments. (103), e52893(2015).</li>
<li>Xie, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((BRD2+compartmentalizes+the+accessible+genome%5BTitle%5D)+AND+%22Nature+Genetics%22%5BJournal%5D)">BRD2 compartmentalizes the accessible genome.</a> Nature Genetics. 54 (4), 481-491 (2022).</li>
<li>Heo, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nuclear+softening+expedites+interstitial+cell+migration+in+fibrous+networks+and+dense+connective+tissues%5BTitle%5D)+AND+%22Science+Advances%22%5BJournal%5D)">Nuclear softening expedites interstitial cell migration in fibrous networks and dense connective tissues.</a> Science Advances. 6 (25), (2020).</li>
<li>Gelléri, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((True-to-scale+DNA-density+maps+correlate+with+major+accessibility+differences+between+active+and+inactive+chromatin%5BTitle%5D)+AND+%22bioRxiv%22%5BJournal%5D)">True-to-scale DNA-density maps correlate with major accessibility differences between active and inactive chromatin.</a> bioRxiv. , (2022).</li>
<li>Derenzini, M., Olins, A. L., Olins, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chromatin+structure+in+situ%3A+the+contribution+of+DNA+ultrastructural+cytochemistry%5BTitle%5D)+AND+%22European+Journal+of+Histochemistry%22%5BJournal%5D)">Chromatin structure in situ: the contribution of DNA ultrastructural cytochemistry.</a> European Journal of Histochemistry. 58 (1), 2307(2014).</li>
<li>Cremer, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+4D+nucleome%3A+Evidence+for+a+dynamic+nuclear+landscape+based+on+co-aligned+active+and+inactive+nuclear+compartments%5BTitle%5D)+AND+%22FEBS+Letters%22%5BJournal%5D)">The 4D nucleome: Evidence for a dynamic nuclear landscape based on co-aligned active and inactive nuclear compartments.</a> FEBS Letters. 589, 2931-2943 (2015).</li>
<li>Shah, F. A., Johansson, B. R., Thomsen, P., Palmquist, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ultrastructural+evaluation+of+shrinkage+artefacts+induced+by+fixatives+and+embedding+resins+on+osteocyte+processes+and+pericellular+space+dimensions%5BTitle%5D)+AND+%22Journal+of+Biomedical+Materials+Research+A%22%5BJournal%5D)">Ultrastructural evaluation of shrinkage artefacts induced by fixatives and embedding resins on osteocyte processes and pericellular space dimensions.</a> Journal of Biomedical Materials Research A. 103 (4), 1565-1576 (2015).</li>
<li>Gavelis, G. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dinoflagellate+nucleus+contains+an+extensive+endomembrane+network%2C+the+nuclear+net%5BTitle%5D)+AND+%22Scientific+Reports%22%5BJournal%5D)">Dinoflagellate nucleus contains an extensive endomembrane network, the nuclear net.</a> Scientific Reports. 9 (1), 839(2019).</li>
<li>Rouquette, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Revealing+the+high-resolution+three-dimensional+network+of+chromatin+and+interchromatin+space%3A+a+novel+electron-microscopic+approach+to+reconstructing+nuclear+architecture%5BTitle%5D)+AND+%22Chromosome+Research%22%5BJournal%5D)">Revealing the high-resolution three-dimensional network of chromatin and interchromatin space: a novel electron-microscopic approach to reconstructing nuclear architecture.</a> Chromosome Research. 17 (6), 801-810 (2009).</li>
<li>Gorisch, S. M., Richter, K., Scheuermann, M. O., Herrmann, H., Lichter, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Diffusion-limited+compartmentalization+of+mammalian+cell+nuclei+assessed+by+microinjected+macromolecules%5BTitle%5D)+AND+%22Experimental+Cell+Research%22%5BJournal%5D)">Diffusion-limited compartmentalization of mammalian cell nuclei assessed by microinjected macromolecules.</a> Experimental Cell Research. 289 (2), 282-294 (2003).</li>
<li>Lenart, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nuclear+envelope+breakdown+in+starfish+oocytes+proceeds+by+partial+NPC+disassembly+followed+by+a+rapidly+spreading+fenestration+of+nuclear+membranes%5BTitle%5D)+AND+%22The+Journal+of+Cell+Biology%22%5BJournal%5D)">Nuclear envelope breakdown in starfish oocytes proceeds by partial NPC disassembly followed by a rapidly spreading fenestration of nuclear membranes.</a> The Journal of Cell Biology. 160 (7), 1055-1068 (2003).</li>
<li>Verschure, P. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Condensed+chromatin+domains+in+the+mammalian+nucleus+are+accessible+to+large+macromolecules%5BTitle%5D)+AND+%22EMBO+Reports%22%5BJournal%5D)">Condensed chromatin domains in the mammalian nucleus are accessible to large macromolecules.</a> EMBO Reports. 4 (9), 861-866 (2003).</li>
<li>Strickfaden, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Condensed+chromatin+behaves+like+a+solid+on+the+mesoscale+in+vitro+and+in+living+cells%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Condensed chromatin behaves like a solid on the mesoscale in vitro and in living cells.</a> Cell. 183 (7), 1772-1784 (2020).</li>
<li>Lin, Y. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((M33+condenses+chromatin+through+nuclear+body+formation+and+methylation+of+both+histone+H3+lysine+9+and+lysine+27%5BTitle%5D)+AND+%22Biochimica+et+Biophysica+Acta.+Molecular+Cell+Research%22%5BJournal%5D)">M33 condenses chromatin through nuclear body formation and methylation of both histone H3 lysine 9 and lysine 27.</a> Biochimica et Biophysica Acta. Molecular Cell Research. 1868 (11), 119100(2021).</li>
<li>Strickfaden, H., Sharma, A. K., Hendzel, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+charge-dependent+phase+transition+determines+interphase+chromatin+organization%5BTitle%5D)+AND+%22bioRxiv%22%5BJournal%5D)">A charge-dependent phase transition determines interphase chromatin organization.</a> bioRxiv. , (2019).</li>
</ol>

<table><tbody><tr><td>Celcultuur</td><td></td><td></td><td></td></tr><tr><td>&micro;-Schaal 35 mm, hoge Grid-500 Glasbodem</td><td>Ibidi</td><td>81168</td><td></td></tr><tr><td>C3H 10T1/2</td><td>IMB (Niehrs Lab)</td><td></td><td></td></tr><tr><td>DMEM</td><td>ThermoFisher</td><td>12320032</td><td></td></tr><tr><td>dPBS</td><td>ThermoFisher</td><td>14190144</td><td></td></tr><tr><td>FBS</td><td>Life Technologies</td><td>16000-044</td><td></td></tr><tr><td>HeLa</td><td>Microscopy Core Facility (IMB)</td><td></td><td></td></tr><tr><td>HFB</td><td>IMB (Kubben Lab)</td><td></td><td></td></tr><tr><td>L-Glutamine</td><td>Sigma-Aldrich</td><td>G7513</td><td></td></tr><tr><td>Niet-essentiële aminozuren en vitamines voor HFB</td><td></td><td></td><td></td></tr><tr><td>Natriumpyruvaat</td><td>S8636</td><td></td><td></td></tr><tr><td>Steekproefvoorbereiding</td><td></td><td></td><td></td></tr><tr><td>Catalase</td><td>Merck</td><td>2593710</td><td></td></tr><tr><td>Glucose</td><td>ThermoFisher</td><td>241922500</td><td></td></tr><tr><td>Glucose-oxidase</td><td>Merck</td><td>49180</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma-Aldrich</td><td>158127</td><td></td></tr><tr><td>RNase Cocktail</td><td>ThermoFisher</td><td>AM2286</td><td></td></tr><tr><td>SYTOX Orange</td><td>ThermoFisher</td><td>S11368</td><td></td></tr><tr><td>TetraSpeck Fluorescent Microspheres Sampler Kit</td><td>ThermoFisher</td><td>T7284</td><td></td></tr><tr><td>Triton X-100</td><td>ThermoFisher</td><td>327372500</td><td></td></tr><tr><td>Software</td><td></td><td></td><td></td></tr><tr><td>BioFormats</td><td>OpenMicroscopy.org</td><td></td><td>open source software https://www.openmicroscopy.org/bio-formats/</td></tr><tr><td>CellProfiler v4.2.1</td><td>CellProfiler.org</td><td></td><td>open source software https://cellprofiler.org</td></tr><tr><td>FiJI</td><td>nih.gov</td><td></td><td>open source software https://imagej.net/software/fiji/?Downloads</td></tr><tr><td>LAND</td><td>nih.gov</td><td></td><td>open source software https://github.com/Jan-NM/LAND</td></tr><tr><td>MatLab 2021</td><td>Math Works</td><td></td><td>commerciële software - vereist "Image Processing Toolbox"</td></tr><tr><td>R v.4..0.3</td><td>r-project.org</td><td></td><td>open source software https://www.r-project.org</td></tr><tr><td>ThunderSTORM v1.3</td><td></td><td></td><td>open source software https://zitmen.github.io/thunderstorm/</td></tr><tr><td>Microscopen:</td><td></td><td></td><td></td></tr><tr><td>AF 7000</td><td>Leica</td><td></td><td></td></tr><tr><td>Leica GSD</td><td>Leica</td><td></td><td></td></tr><tr><td>SMLM Microscoop</td><td>Cremer Lab</td><td></td><td>op maat gebouwd door Dr. S-Y. Chen</td></tr></tbody></table>

absolute dna-dichtheid in celkernen in kaart brengen met behulp van lokalisatiemicroscopie met één molecuul

Mapping Absolute DNA Density in Cell Nuclei using Single-molecule Localization Microscopy

Absolute DNA-dichtheid in celkernen in kaart brengen met behulp van lokalisatiemicroscopie met één molecuul

Within the cell nucleus, silent genes are generally located in chromatin areas of high density called heterochromatin, whereas active genes can be mostly ...

mapping-absolute-dna-density-cell-nuclei-using-single-molecule

Research

JoVE Journal

Biology

Biologie

Enkel molecuul microscopie Approach die nieuwe inzichten naar nucleaire transport.

Single-molecule beeldvorming van de nucleaire transporten

We beschrijven een robuuste en veelzijdige protocol voor de analyse nucleaire architectuur door 3D DNA FISH met direct gelabelde fluorescerende probes.

Robuuste 3D DNA FISH behulp direct gemerkt Probes

Fotogeactiveerde lokalisatie microscopie (PALM) in combinatie met single-molecule-tracking maakt directe observatie en kwantificering van eiwit-DNA-interacties in levende Escherichia coli cellen.

Visualiseren eiwit-DNA-interacties in levende bacteriële cellen met behulp Photoactivated Enkelmolecuul Tracking

Gepresenteerd is een flexibele informatica workflow waardoor multiplex-image-based analyse van fluorescent gelabelde cellen. De workflow kwantificeert nucleaire en cytoplasmatische markers en berekent marker translocatie tussen deze compartimenten. Procedures zijn voorzien verstoring van cellen met siRNA en betrouwbare methode voor het merkerdetectie door indirecte immunofluorescentie in 96-well formaat.

Workflow voor High-inhoud, individuele cel Kwantificering van fluorescente merkers van Universal Microscoop gegevens, Ondersteund door Open Source Software

Onlangs ontdekte geoxideerde vormen van 5-methylcytosine (oxi-mCs), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) en 5-carboxylcytosine (5caC) kan verschillende DNA wijzigingen met unieke functionele rollen. Hier wordt een semi-kwantitatieve workflow voor visualisatie van oxi-mCs ruimtelijke verdeling, signaal intensiteit profilering en colocalization omschreven.

Immunokleuring voor DNA wijzigingen: computationele analyse van Confocal beelden

3D single-molecuul lokalisatie microscopie wordt gebruikt om de ruimtelijke posities en bewegings trajecten van fluorescently gelabelde eiwitten in levende bacteriële cellen te sonde. Het experimentele en data-analyse protocol dat hierin wordt beschreven, bepaalt het heersende diffusieve gedrag van cytosolische eiwitten op basis van gegroepeerde enkelvoudige molecuul trajecten.

Single-molecule tracking microscopie-een hulpmiddel voor het bepalen van de Diffusieve toestanden van cytosolische moleculen

3D multicolor DNA FISH vertegenwoordigt een hulpmiddel om meerdere genomische loci binnen 3D bewaarde kernen te visualiseren, ondubbelzinnig definiëren d.m.v. hun wederzijdse interacties en lokalisatie binnen de nucleaire ruimte op één celniveau. Hier wordt een stap voor stap protocol beschreven voor een breed spectrum van menselijke primaire cellen.

3D Multicolor DNA FISH Tool om nucleaire architectuur in menselijke primaire cellen te bestuderen

Dit protocol beschrijft single-antibody labeling (SAL) om de nanoschaal ruimtelijke organisatie van plasmamembraaneiwitten op te lossen. Door cumulatieve antilichaam-epitoopinteracties op het niveau van één molecuul te benutten, brengt membraan SAL (mSAL) lokale epitoopverdelingen in kaart terwijl het tegelijkertijd het bindingsgedrag van antilichamen in de natuurlijke cellulaire omgeving vastlegt.

Single-molecule localisatiemicroscopie van membraaneiwitten met single-antibody labeling

We presenteren een driekleurig chromatine single molecule localization microscopy (SMLM) kleurings- en analyseprotocol dat reproduceerbare mapping van euchromatine, heterochromatine en RNAP II mogelijk maakt voor ruimtelijke analyse. Dit protocol maakt efficiënte meerkleurige etikettering mogelijk in dichte nucleaire omgevingen, inclusief chromatine-geassocieerde doelen, waardoor betrouwbare gelijktijdige detectie mogelijk is.

Een multilabel Single Molecule Localization Microscopieprotocol voor het Onderzoek van Chromatine in de dichte nucleaire omgeving

Dit protocol biedt gedetailleerde bewerkingen voor monstervoorbereiding, gegevensverzameling en analyse van DNA intrinsieke fluorescentiespectra.

Spectroscopische superresolutiebeeldvorming van DNA-moleculen met intrinsiek contrast

Dit protocol biedt een efficiënte workflow voor single-molecule localisatie microscopiebeeldvorming van geïsoleerde Arabidopsis-kernen, waarbij geoptimaliseerde isolatie, fixatie en labeling wordt gebruikt om betrouwbare nanoschaalvisualisatie van chromatine en RNA Polymerase II te bereiken. Deze benadering kan gemakkelijk worden aangepast aan andere chromatine-geassocieerde eiwitten of histonmodificaties voor beeldvorming met hoge resolutie.

Een gestroomlijnd protocol voor single-molecule localisatiemicroscopie in Arabidopsis-kernen

Leer absolute DNA-dichtheid meten in celkernen met behulp van Voronoi-tessellatie en single-molecule localisatiemicroscopie. Deze methode biedt inzicht in chromatine-architectuur en ruimtelijke beperkingen.

Absolute DNA-dichtheid in celkernen in kaart brengen met behulp van lokalisatiemicroscopie met één molecuul

Meer video's verkennen