Name: İçsel Kontrast kullanarak DNA moleküllerinin spektroskopik süper çözünürlüklü görüntülemesi (Video) | JoVE Journal
Uploaded: 2026-03-06T13:25:00.000+00:00
Duration: 9 min 19 s
Description: Öğrenin, DNA moleküllerinin içsel floresansını kullanarak etiketsiz süper çözünürlüklü spektroskopik görüntüleme protokolünü. Bu yöntem, yerel optik özellikleri koruyarak yüksek çözünürlüklü analiz sağlar.

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<ol><li>Ellis, R. J., Minton, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Join+the+crowd%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Join the crowd.</a> Nature. 425 (6953), 27-28 (2003).</li>
<li>Hell, S. W., Wichmann, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Breaking+the+diffraction+resolution+limit+by+stimulated+emission%3A+stimulated+emission-depletion+fluorescence+microscopy%5BTitle%5D)+AND+%22Opt+Lett%22%5BJournal%5D)">Breaking the diffraction resolution limit by stimulated emission: stimulated emission-depletion fluorescence microscopy.</a> Opt Lett. 19 (11), 780(1994).</li>
<li>Rust, M. J., Bates, M., Zhuang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sub-diffraction-limit+imaging+by+stochastic+optical+reconstruction+microscopy+%28STORM%29%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).</a> Nat Methods. 3 (10), 793-796 (2006).</li>
<li>Betzig, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Imaging+intracellular+fluorescent+proteins+at+nanometer+resolution%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Imaging intracellular fluorescent proteins at nanometer resolution.</a> Science. 313 (5793), 1642-1645 (2006).</li>
<li>Gustafsson, M. G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nonlinear+structured-illumination+microscopy%3A+wide-field+fluorescence+imaging+with+theoretically+unlimited+resolution%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution.</a> Proc Natl Acad Sci U S A. 102 (37), 13081-13086 (2005).</li>
<li>Hell, S. W., Kroug, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ground-state-depletion+fluorescence+microscopy%3A+a+concept+for+breaking+the+diffraction+resolution+limit%5BTitle%5D)+AND+%22Appl+Phys+B%22%5BJournal%5D)">Ground-state-depletion fluorescence microscopy: a concept for breaking the diffraction resolution limit.</a> Appl Phys B. 60 (5), 495-497 (1995).</li>
<li>Fölling, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fluorescence+nanoscopy+by+ground-state+depletion+and+single-molecule+return%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">Fluorescence nanoscopy by ground-state depletion and single-molecule return.</a> Nat Methods. 5 (11), 943-945 (2008).</li>
<li>Ricci, M., Manzo, C., García-Parajo, M. F., Lakadamyali, M., Cosma, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Chromatin+fibers+are+formed+by+heterogeneous+groups+of+nucleosomes+in+vivo%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Chromatin fibers are formed by heterogeneous groups of nucleosomes in vivo.</a> Cell. 160 (6), 1145-1158 (2015).</li>
<li>Bintu, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Super-resolution+chromatin+tracing+reveals+domains+and+cooperative+interactions+in+single+cells%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Super-resolution chromatin tracing reveals domains and cooperative interactions in single cells.</a> Science. 362 (6413), eaau1783(2018).</li>
<li>Matsuda, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Condensed+mitotic+chromosome+structure+at+nanometer+resolution+using+PALM+and+EGFP-histones%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Condensed mitotic chromosome structure at nanometer resolution using PALM and EGFP-histones.</a> PLoS One. 5 (9), e12768(2010).</li>
<li>Bohn, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Localization+microscopy+reveals+expression-dependent+parameters+of+chromatin+nanostructure%5BTitle%5D)+AND+%22Biophys+J%22%5BJournal%5D)">Localization microscopy reveals expression-dependent parameters of chromatin nanostructure.</a> Biophys J. 99 (5), 1358-1367 (2010).</li>
<li>Acosta, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Three-color+single-molecule+localization+microscopy+in+chromatin%5BTitle%5D)+AND+%22Light+Sci+Appl%22%5BJournal%5D)">Three-color single-molecule localization microscopy in chromatin.</a> Light Sci Appl. 14, 7(2025).</li>
<li>Almassalha, L. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Label-free+imaging+of+the+native%2C+living+cellular+nanoarchitecture+using+partial-wave+spectroscopic+microscopy%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Label-free imaging of the native, living cellular nanoarchitecture using partial-wave spectroscopic microscopy.</a> Proc Natl Acad Sci U S A. 113 (42), E6372-E6381 (2016).</li>
<li>Beerman, T. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+analogs+of+the+DNA+minor+groove+binder+Hoechst+33258+on+topoisomerase+II+and+I+mediated+activities%5BTitle%5D)+AND+%22Biochim+Biophys+Acta%22%5BJournal%5D)">Effects of analogs of the DNA minor groove binder Hoechst 33258 on topoisomerase II and I mediated activities.</a> Biochim Biophys Acta. 1131 (1), 53-61 (1992).</li>
<li>Cadet, J., Anselmino, C., Douki, T., Voituriez, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+trends+in+photobiology%5BTitle%5D)+AND+%22J+Photochem+Photobiol+B%22%5BJournal%5D)">New trends in photobiology.</a> J Photochem Photobiol B. 15 (4), 277-298 (1992).</li>
<li>Dong, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Superresolution+intrinsic+fluorescence+imaging+of+chromatin+utilizing+native%2C+unmodified+nucleic+acids+for+contrast%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Superresolution intrinsic fluorescence imaging of chromatin utilizing native, unmodified nucleic acids for contrast.</a> Proc Natl Acad Sci U S A. 113 (35), 9716-9721 (2016).</li>
<li>Vayá, I., Gustavsson, T., Miannay, F., Douki, T., Markovitsi, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fluorescence+of+natural+DNA%3A+from+the+femtosecond+to+the+nanosecond+time+scales%5BTitle%5D)+AND+%22J+Am+Chem+Soc%22%5BJournal%5D)">Fluorescence of natural DNA: from the femtosecond to the nanosecond time scales.</a> J Am Chem Soc. 132 (34), 11834-11835 (2010).</li>
<li>Anders, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DNA+fluorescence+at+room+temperature+excited+by+means+of+a+dye+laser%5BTitle%5D)+AND+%22Chem+Phys+Lett%22%5BJournal%5D)">DNA fluorescence at room temperature excited by means of a dye laser.</a> Chem Phys Lett. 81 (2), 270-272 (1981).</li>
<li>Plessow, R., Brockhinke, A., Eimer, W., Kohse-Höinghaus, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intrinsic+time-+and+wavelength-resolved+fluorescence+of+oligonucleotides%3A+a+systematic+investigation+using+a+novel+picosecond+laser+approach%5BTitle%5D)+AND+%22J+Phys+Chem+B%22%5BJournal%5D)">Intrinsic time- and wavelength-resolved fluorescence of oligonucleotides: a systematic investigation using a novel picosecond laser approach.</a> J Phys Chem B. 104 (15), 3695-3704 (2000).</li>
<li>Takaya, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((UV+excitation+of+single+DNA+and+RNA+strands+produces+high+yields+of+exciplex+states+between+two+stacked+bases%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">UV excitation of single DNA and RNA strands produces high yields of exciplex states between two stacked bases.</a> Proc Natl Acad Sci U S A. 105 (30), 10285-10290 (2008).</li>
<li>Bancaud, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+fractal+model+for+nuclear+organization%3A+current+evidence+and+biological+implications%5BTitle%5D)+AND+%22Nucleic+Acids+Res%22%5BJournal%5D)">A fractal model for nuclear organization: current evidence and biological implications.</a> Nucleic Acids Res. 40 (18), 8783-8792 (2012).</li>
<li>Daban, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High+concentration+of+DNA+in+condensed+chromatin%5BTitle%5D)+AND+%22Biochem+Cell+Biol%22%5BJournal%5D)">High concentration of DNA in condensed chromatin.</a> Biochem Cell Biol. 81 (3), 91-99 (2003).</li>
<li>Bohrmann, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Concentration+evaluation+of+chromatin+in+unstained+resin-embedded+sections+by+means+of+low-dose+ratio-contrast+imaging+in+STEM%5BTitle%5D)+AND+%22Ultramicroscopy%22%5BJournal%5D)">Concentration evaluation of chromatin in unstained resin-embedded sections by means of low-dose ratio-contrast imaging in STEM.</a> Ultramicroscopy. 49 (1-4), 235-251 (1993).</li>
<li>Dong, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Stochastic+fluorescence+switching+of+nucleic+acids+under+visible+light+illumination%5BTitle%5D)+AND+%22Opt+Express%22%5BJournal%5D)">Stochastic fluorescence switching of nucleic acids under visible light illumination.</a> Opt Express. 25 (7), 7929-7944 (2017).</li>
<li>Gong, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DNA+spectroscopic+intrinsic-contrast+photon-localization+optical+nanoscopy%5BTitle%5D)+AND+%22Proc+SPIE%22%5BJournal%5D)">DNA spectroscopic intrinsic-contrast photon-localization optical nanoscopy.</a> Proc SPIE. 13326, 133260F(2025).</li>
<li>Wang, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Label-free+DNA+spectroscopic+photon-localization+nanoscopy%5BTitle%5D)+AND+%22Optica+Biophotonic+Congress%22%5BJournal%5D)">Label-free DNA spectroscopic photon-localization nanoscopy.</a> Optica Biophotonic Congress. , (2025).</li>
<li>Eshein, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sub-10-nm+imaging+of+nucleic+acids+using+spectroscopic+intrinsic-contrast+photon-localization+optical+nanoscopy+%28SICLON%29%5BTitle%5D)+AND+%22Opt+Lett%22%5BJournal%5D)">Sub-10-nm imaging of nucleic acids using spectroscopic intrinsic-contrast photon-localization optical nanoscopy (SICLON).</a> Opt Lett. 43 (23), 5817-5820 (2018).</li>
<li>Song, K. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Monolithic+dual-wedge+prism-based+spectroscopic+single-molecule+localization+microscopy%5BTitle%5D)+AND+%22Nanophotonics%22%5BJournal%5D)">Monolithic dual-wedge prism-based spectroscopic single-molecule localization microscopy.</a> Nanophotonics. 11 (8), 1527-1535 (2022).</li>
<li>Song, K. H., Sun, C., Zhang, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Design+strategy+for+a+dual-wedge+prism+imaging+spectrometer+in+spectroscopic+nanoscopy%5BTitle%5D)+AND+%22Rev+Sci+Instrum%22%5BJournal%5D)">Design strategy for a dual-wedge prism imaging spectrometer in spectroscopic nanoscopy.</a> Rev Sci Instrum. 94 (2), 023702(2023).</li>
<li>Yeo, W. H., Sun, C., Zhang, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Physically+informed+Monte+Carlo+simulation+of+dual-wedge+prism-based+spectroscopic+single-molecule+localization+microscopy%5BTitle%5D)+AND+%22J+Biomed+Opt%22%5BJournal%5D)">Physically informed Monte Carlo simulation of dual-wedge prism-based spectroscopic single-molecule localization microscopy.</a> J Biomed Opt. 29 (S1), S11501(2023).</li>
<li>Davis, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((RainbowSTORM%3A+an+open-source+ImageJ+plug-in+for+spectroscopic+single-molecule+localization+microscopy+%28sSMLM%29+data+analysis+and+image+reconstruction%5BTitle%5D)+AND+%22Bioinformatics%22%5BJournal%5D)">RainbowSTORM: an open-source ImageJ plug-in for spectroscopic single-molecule localization microscopy (sSMLM) data analysis and image reconstruction.</a> Bioinformatics. 36 (19), 4972-4974 (2020).</li>
<li>Davis, J. L., et al. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aDavis%2C+JL+%22RainbowSTORM+v2+%28MATLAB+app%29%22">RainbowSTORM v2 (MATLAB app)</a>. , https://github.com/FOIL-NU/sSMLM-protocol (2020).</li>
<li>Brenner, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Implementation+and+calibration+of+spectroscopic+single-molecule+localization+microscopy%5BTitle%5D)+AND+%22BMC+Methods%22%5BJournal%5D)">Implementation and calibration of spectroscopic single-molecule localization microscopy.</a> BMC Methods. 2 (1), 2(2025).</li>
<li>sSMLM for DNA intrinsic fluorescence (Python scripts). , Backman Lab. 
 <a target="_blank" href="https://github.com/BackmanLab/sSMLM-for-DNA-intrinsic-fluorescence">https://github.com/BackmanLab/sSMLM-for-DNA-intrinsic-fluorescence</a> (2025).</li>
<li>N-STORM protocol. , Molecular Vista. Available from: 
 <a target="_blank" href="https://www.mvi-inc.com/wp-content/uploads/N-STORM+Protocol.pdf">https://www.mvi-inc.com/wp-content/uploads/N-STORM+Protocol.pdf</a> (2014).</li>
<li>Abdelsayed, V., Boukhatem, H., Olivier, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+optimized+buffer+for+repeatable+multicolor+STORM%5BTitle%5D)+AND+%22ACS+Photonics%22%5BJournal%5D)">An optimized buffer for repeatable multicolor STORM.</a> ACS Photonics. 9 (12), 3926-3934 (2022).</li>
<li>Zumbusch, A., Holtom, G. R., Xie, X. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Three-dimensional+vibrational+imaging+by+coherent+anti-Stokes+Raman+scattering%5BTitle%5D)+AND+%22Phys+Rev+Lett%22%5BJournal%5D)">Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering.</a> Phys Rev Lett. 82 (20), 4142(1999).</li>
<li>Benninger, R. K. P., Piston, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Two%E2%80%90photon+excitation+microscopy+for+the+study+of+living+cells+and+tissues%5BTitle%5D)+AND+%22Curr+Protoc+Cell+Biol%22%5BJournal%5D)">Two‐photon excitation microscopy for the study of living cells and tissues.</a> Curr Protoc Cell Biol. 59 (1), 4-11 (2013).</li>
<li>Zhang, C., Zhang, D., Cheng, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coherent+Raman+scattering+microscopy+in+biology+and+medicine%5BTitle%5D)+AND+%22Ann+Rev+Biomed+Eng%22%5BJournal%5D)">Coherent Raman scattering microscopy in biology and medicine.</a> Ann Rev Biomed Eng. 17 (1), 415-445 (2015).</li>
<li>Song, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SNR+enhanced+high-speed+two-photon+microscopy+using+a+pulse+picker+and+time+gating+detection%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">SNR enhanced high-speed two-photon microscopy using a pulse picker and time gating detection.</a> Sci Rep. 13 (1), 14244(2023).</li>
<li>Luu, P., Fraser, S. E., Schneider, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((More+than+double+the+fun+with+two-photon+excitation+microscopy%5BTitle%5D)+AND+%22Comm+Biol%22%5BJournal%5D)">More than double the fun with two-photon excitation microscopy.</a> Comm Biol. 7 (1), 364(2024).</li>
<li>Van de Linde, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Direct+stochastic+optical+reconstruction+microscopy+with+standard+fluorescent+probes%5BTitle%5D)+AND+%22Nat+Protoc%22%5BJournal%5D)">Direct stochastic optical reconstruction microscopy with standard fluorescent probes.</a> Nat Protoc. 6 (7), 991-1009 (2011).</li>
<li>Wang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Research+progress+on+the+luminescence+of+biomacromolecules%5BTitle%5D)+AND+%22J+Mater+Sci+Technol%22%5BJournal%5D)">Research progress on the luminescence of biomacromolecules.</a> J Mater Sci Technol. 76, 60-75 (2021).</li>
<li>Varejão, N., Reverter, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Using+intrinsic+fluorescence+to+measure+protein+stability+upon+thermal+and+chemical+denaturation%5BTitle%5D)+AND+%22Methods+Mol+Biol%22%5BJournal%5D)">Using intrinsic fluorescence to measure protein stability upon thermal and chemical denaturation.</a> Methods Mol Biol. 2581, 229-241 (2023).</li>
<li>Sindrewicz, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intrinsic+tryptophan+fluorescence+spectroscopy+reliably+determines+galectin-ligand+interactions%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Intrinsic tryptophan fluorescence spectroscopy reliably determines galectin-ligand interactions.</a> Sci Rep. 9 (1), 11851(2019).</li>
<li>Ho, C. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rapid+identification+of+pathogenic+bacteria+using+Raman+spectroscopy+and+deep+learning%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Rapid identification of pathogenic bacteria using Raman spectroscopy and deep learning.</a> Nat Commun. 10 (1), 4927(2019).</li>
</ol>

<table><tbody><tr><td>görüntüleme için 488 nm emisyon filtresi</td><td>Chroma Technology Corp.</td><td>ET525/50M</td><td></td></tr><tr><td>Kalibrasyon için 532 nm bant geçirici filtre</td><td>Thorlabs</td><td>FL532-3</td><td></td></tr><tr><td>görüntüleme için 532 nm emisyon filtresi</td><td>Chroma Technology Corp.</td><td>ET620/60M</td><td></td></tr><tr><td>Kalibrasyon için 580 nm bant geçişli filtre</td><td>Thorlabs</td><td>FB580-10</td><td></td></tr><tr><td>Kalibrasyon için 633 nm bant geçişli filtre</td><td>Thorlabs</td><td>FL632.8-3</td><td></td></tr><tr><td>Kalibrasyon için 680 nm bant geçişli filtre</td><td>Thorlabs</td><td>FB680-10</td><td></td></tr><tr><td>Kalibrasyon için 750 nm bant geçişli filtre</td><td>Thorlabs</td><td>FB750-10</td><td></td></tr><tr><td>Beta-merkaptoetanol&nbsp;</td><td>Sigma</td><td>M6250</td><td></td></tr><tr><td>BME görüntüleme tamponu</td><td>NA</td><td>NA</td><td>7 &mu; L GLOX, 7 &mu; L Beta-merkaptoetanol ve 690 &mu; L glikoz tamponu</td></tr><tr><td>Sığır karaciğerinden katalaz</td><td>Sigma</td><td>C100</td><td></td></tr><tr><td>DABCO (1,4-di-azobisicyclo-(2.2.2.)-oktan)</td><td>Sigma</td><td>D27802</td><td></td></tr><tr><td>DABCO görüntüleme tamponu</td><td>NA</td><td>NA</td><td>65 mM DABCO+30 mM Sodyum sülfit + 30 mM DTT 1 M&nbsp; DNAse'de, RNAse serbest deionize su. Sodyum sülfit PBS ile 10 kez çözünmelidir; görüntüleme tamponunu oluşturmadan önce 1 M'ye kadar devam eder.</td></tr><tr><td>Spektroskopik SMLM için DNA deplexi</td><td>IDT</td><td>NA</td><td>20 tabanlı çift çift zincirli GC_alter, GC_conti, AT_alter ve AT_conti. Örneğin GC_alter her iki iplik de 5' olarak - GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCCGCGCGCCGCGCGCGCGCGCGCG GC_conti bir ipliği 5' uzunluğunda - GGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG diğeri ise 5' olarak - CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC - 3' Tüm örnekler HPLC standardındadır.</td></tr><tr><td>DNA oligonukleotid/çift zincirli DNA</td><td>IDT</td><td>NA</td><td></td></tr><tr><td>Spektroskopik SMLM için DNA oligoları</td><td>IDT</td><td>NA</td><td>20 tabanlı A, G, C, T; 40 tabanlı tek zincirli AC zinciri (A1C1, A5C5, A10C10, A20C20); &nbsp; tek zincirli 5-, 8-, 12-, 16-bazlı Guanine. Örneğin A10C10 is 5'- AAAAAAAACCCCC CCCCCAAAAAAAA AACCCCCCCCCC - 3' Tüm örnekler HPLC standardındadır.</td></tr><tr><td>DNaz/RNase-Serbest Damıtılmış Su</td><td>Invitrogen</td><td>10977015</td><td></td></tr><tr><td>DTT (DL-Ditiothreitol)</td><td>Sigma</td><td>43816</td><td></td></tr><tr><td>GLOX</td><td>NA</td><td>NA</td><td>14 mg glikoz oksidaz, 50 &mu; L katalaz (17 mg/ml) + 200 &mu; L tampon A, yani 10 mM Tris (pH 8.0) ve 50 mM NaCl.&nbsp;</td></tr><tr><td>Glikoz tamponu</td><td>NA</td><td>NA</td><td>50 mM Tris (pH 8.0), 50 mM NaCl ve %10 glikoz.&nbsp;</td></tr><tr><td>Glukoz oksidaz</td><td>Sigma</td><td>G7141</td><td></td></tr><tr><td>Glikoz, toz</td><td>Thermo Fisher</td><td>15023021</td><td></td></tr><tr><td>PBS (10X)</td><td>Thermo Fisher</td><td>J62036. K3</td><td></td></tr><tr><td>PBS (1X)</td><td>Gibco</td><td>10010023</td><td></td></tr><tr><td>Sodyum klorür</td><td>Thermo Fisher</td><td>424290010</td><td></td></tr><tr><td>Sodyum sülfit&nbsp;</td><td>Sigma</td><td>S0505</td><td></td></tr><tr><td>Tris (1M) pH 8.0</td><td>Invitrogen</td><td>AM9856</td><td></td></tr></tbody></table>

İçsel kontrast kullanarak dna moleküllerinin spektroskopik süper çözünürlüklü görüntülemesi

Spectroscopic Super-resolution Imaging of DNA Molecules using Intrinsic Contrast

İçsel Kontrast kullanarak DNA moleküllerinin spektroskopik süper çözünürlüklü görüntülemesi

Super-resolution imaging has revolutionized biological research by revealing structural details at the nanoscale. Most optical super-resolution methods ...

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Bioengineering

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Bu videoda, tek moleküllü görüntüleme kullanarak belirli lokuslarda DNA molekülleri üzerinde füzyon proteini yoğunlaşmalarının montajını görselleştirmek için bir yöntem açıklanmaktadır. Bu kondensatlar, RNA polimeraz II'nin işe alınmasına ve gelişmiş gen transkripsiyonuna izin veren LCD-LCD ve LCD-DBD etkileşimleri nedeniyle oluşur.

DNA perdeleri üzerinde füzyon proteini kondensatlarının montajını görselleştirmek için tek moleküllü görüntüleme

Biz bakteriyel FtsZ-ring yapısal organizasyonu, hücre bölünmesi için gerekli bir aparat soruşturma için görüntüleme yöntemi bir süper-çözünürlük tarif. Bu yöntem, fotoaktive yerelleştirme mikroskobu (PALM) kantitatif görüntü analizi dayanır ve diğer bakteri sitoskeletal proteinleri ile uygulanabilir.

Bakteriyel Bölümü Makine Super çözünürlüklü Görüntüleme

Yeni 5-metilsitozin (oksi-mCs), 5-arasında bakterilerde görülen (5hmC), 5-formylcytosine (5 fC) oksitlenmiş formları bulunan ve 5-carboxylcytosine (5caC) farklı DNA değişiklikleri benzersiz işlevsel roller ile temsil edebilir. Yarı nicel bir iş akışı görselleştirme oksi-mCs kayma dağıtım, sinyal şiddeti profil oluşturma ve colocalization için burada kullanılmıştır.

DNA değişiklikler için Immunostaining: Confocal görüntülerin sayısal analiz

Bu metodoloji genel amacı en iyi deneysel koşullar numune hazırlama resim alma ve yeniden yapılanma için 2D çift renk dSTORM görüntülerini mikrotübüller ve ara filamentler sabit hücrelerde gerçekleştirmek için...

Ara filamentler ve mikrotübüller 2 boyutlu doğrudan Stokastik optik imar mikroskobu ile görüntüleme

Ayrıntılı bir talimat nasıl son derece eğimli kurmak için kiremit (HIST) mikroskop ve onun kullanım için tek molekül düşsel süpürüldü açıklanmıştır.

Son derece eğimli oluşturmak için bir kılavuz döşeme mikroskop genişletilmiş görünümü alan tek molekül görüntüleme için süpürüldü.

3D çok renkli DNA FISH, 3B korunmuş çekirdekler içinde birden fazla genomik loci görselleştirmek için bir araç temsil eder, kesin olarak tek bir hücre düzeyinde nükleer alan içinde karşılıklı etkileşimleri ve lokalizasyon tanımlayan. Burada, adım protokol bir adım insan birincil hücrelerin geniş bir yelpazede için açıklanmıştır.

3D Çok Renkli DNA FISH Aracı İnsan Birincil Hücrelerde Nükleer Mimari Çalışma

Mevcut protokol, tek moleküllü lokalizasyon mikroskobu verilerinin, bilinen hacmin, genom boyutunun ve hücre döngüsü aşamasının Voronoi mozaiklemesini kullanarak yapışık hücre çekirdekleri içindeki mutlak DNA yoğunluklarını ölçen bir yöntemi açıklamaktadır.

Tek Molekül Lokalizasyon Mikroskobu Kullanılarak Hücre Çekirdeğinde Mutlak DNA Yoğunluğunun Haritalanması

Bu protokol, CMG sarmal ile çözülen DNA'nın canlı görselleştirilmesi için tek moleküllü bir testin gerçekleştirilmesini gösterir. (1) bir DNA substratının hazırlanmasını, (2) floresan etiketli Drosophila melanogaster CMG helikazının saflaştırılmasını, (3) toplam iç yansıma floresan (TIRF) mikroskobu için bir mikroakışkan akış hücresinin hazırlanmasını ve (4) tek moleküllü DNA çözme tahlilini açıklar.

CMG Helikaz ile DNA Çözmenin Tek Moleküllü Gerçek Zamanlı Görselleştirilmesi

Bu protokol, optik foton yeniden atamalı olan dönen disk konfokal mikroskobu kullanarak derin, tam hücre tek moleküllü lokalizasyon mikroskopisi (SMLM) uygulanma prosedürünü özetler; böylece özel optik veya karmaşık aydınlatmaya ihtiyaç olmadan DNA-PAINT görüntüleme mümkün olur.

Optik foton yeniden atamalı dönen disk konfokal üzerinde DNA-PAINT ile tüm hücre süper çözünürlüklü görüntüleme

"E-DNA" sensörleri, reagentless, kan ve diğer karmaşık matrisler doğrudan meydan bile iyi bir performans elektrokimyasal biyosensörler, nükleik asit, protein ve küçük molekül analitler geniş bir algılama için adapte edilmiştir. Burada bu tür sensörlerin üretimi ve kullanımı için genel bir prosedür mevcut.

Nükleik Asitler, Proteinler ve Küçük Moleküller Reagentless Algılama Elektrokimyasal DNA Biyosensörler Fabrikasyon

Öğrenin, DNA moleküllerinin içsel floresansını kullanarak etiketsiz süper çözünürlüklü spektroskopik görüntüleme protokolünü. Bu yöntem, yerel optik özellikleri koruyarak yüksek çözünürlüklü analiz sağlar.

İçsel Kontrast kullanarak DNA moleküllerinin spektroskopik süper çözünürlüklü görüntülemesi

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