Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structure
Immunostaining and fluorescence in situ hybridization (FISH) are well established methods for specific labelling of chromatin in the cell nucleus. COMBO-FISH (combinatorial oligonucleotide fluorescence in situ hybridization) is a FISH method using computer designed oligonucleotide probes specificall...
Gespeichert in:
| Hauptverfasser: | , , |
|---|---|
| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
7 May 2017
|
| In: |
International journal of molecular sciences
Year: 2017, Jahrgang: 18, Heft: 5 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms18051005 |
| Online-Zugang: | Verlag, kostenfrei, Volltext: http://dx.doi.org/10.3390/ijms18051005 Verlag, kostenfrei, Volltext: http://www.mdpi.com/1422-0067/18/5/1005 
|
| Verfasserangaben: | Matthias Krufczik, Aaron Sievers, Annkathrin Hausmann, Jin-Ho Lee, Georg Hildenbrand, Wladimir Schaufler and Michael Hausmann |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1560088648 | ||
| 003 | DE-627 | ||
| 005 | 20230428071243.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 170622s2017 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.3390/ijms18051005 |2 doi | |
| 035 | |a (DE-627)1560088648 | ||
| 035 | |a (DE-576)490088643 | ||
| 035 | |a (DE-599)BSZ490088643 | ||
| 035 | |a (OCoLC)1263875883 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 29 |2 sdnb | ||
| 100 | 1 | |a Krufczik, Matthias |d 1986- |e VerfasserIn |0 (DE-588)1076034802 |0 (DE-627)834101076 |0 (DE-576)444809643 |4 aut | |
| 245 | 1 | 0 | |a Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structure |c Matthias Krufczik, Aaron Sievers, Annkathrin Hausmann, Jin-Ho Lee, Georg Hildenbrand, Wladimir Schaufler and Michael Hausmann |
| 264 | 1 | |c 7 May 2017 | |
| 300 | |a 14 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Gesehen am 22.06.2017 | ||
| 520 | |a Immunostaining and fluorescence in situ hybridization (FISH) are well established methods for specific labelling of chromatin in the cell nucleus. COMBO-FISH (combinatorial oligonucleotide fluorescence in situ hybridization) is a FISH method using computer designed oligonucleotide probes specifically co-localizing at given target sites. In combination with super resolution microscopy which achieves spatial resolution far beyond the Abbe Limit, it allows new insights into the nano-scaled structure and organization of the chromatin of the nucleus. To avoid nano-structural changes of the chromatin, the COMBO-FISH labelling protocol was optimized omitting heat treatment for denaturation of the target. As an example, this protocol was applied to ALU elements—dispersed short stretches of DNA which appear in different kinds in large numbers in primate genomes. These ALU elements seem to be involved in gene regulation, genomic diversity, disease induction, DNA repair, etc. By computer search, we developed a unique COMBO-FISH probe which specifically binds to ALU consensus elements and combined this DNA-DNA labelling procedure with heterochromatin immunostainings in formaldehyde-fixed cell specimens. By localization microscopy, the chromatin network-like arrangements of ALU oligonucleotide repeats and heterochromatin antibody labelling sites were simultaneously visualized and quantified. This novel approach which simultaneously combines COMBO-FISH and immunostaining was applied to chromatin analysis on the nanoscale after low-linear-energy-transfer (LET) radiation exposure at different doses. Dose-correlated curves were obtained from the amount of ALU representing signals, and the chromatin re-arrangements during DNA repair after irradiation were quantitatively studied on the nano-scale. Beyond applications in radiation research, the labelling strategy of immunostaining and COMBO-FISH with localization microscopy will also offer new potentials for analyses of subcellular elements in combination with other specific chromatin targets. | ||
| 650 | 4 | |a ALU-repeats | |
| 650 | 4 | |a cell nucleus | |
| 650 | 4 | |a chromatin arrangement | |
| 650 | 4 | |a combinatorial oligonucleotide FISH (COMBO-FISH) | |
| 650 | 4 | |a combined immuno-staining | |
| 650 | 4 | |a localization microscopy | |
| 650 | 4 | |a low LET radiation exposure | |
| 700 | 1 | |a Hildenbrand, Georg Lars |d 1972- |e VerfasserIn |0 (DE-588)1068900059 |0 (DE-627)821002562 |0 (DE-576)428273017 |4 aut | |
| 700 | 1 | |a Hausmann, Michael |d 1966- |e VerfasserIn |0 (DE-588)120185733 |0 (DE-627)696450747 |0 (DE-576)292085257 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t International journal of molecular sciences |d Basel : Molecular Diversity Preservation International, 2000 |g 18(2017,5) Artikel-Nummer 1005, 16 Seiten |h Online-Ressource |w (DE-627)316340715 |w (DE-600)2019364-6 |w (DE-576)281194653 |x 1422-0067 |7 nnas |a Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structure |
| 773 | 1 | 8 | |g volume:18 |g year:2017 |g number:5 |g extent:14 |a Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structure |
| 856 | 4 | 0 | |u http://dx.doi.org/10.3390/ijms18051005 |x Verlag |x Resolving-System |z kostenfrei |3 Volltext |
| 856 | 4 | 0 | |u http://www.mdpi.com/1422-0067/18/5/1005 |x Verlag |z kostenfrei |3 Volltext |
| 951 | |a AR | ||
| 992 | |a 20170622 | ||
| 993 | |a Article | ||
| 994 | |a 2017 | ||
| 998 | |g 120185733 |a Hausmann, Michael |m 120185733:Hausmann, Michael |d 130000 |d 130700 |e 130000PH120185733 |e 130700PH120185733 |k 0/130000/ |k 1/130000/130700/ |p 7 |y j | ||
| 998 | |g 1068900059 |a Hildenbrand, Georg Lars |m 1068900059:Hildenbrand, Georg Lars |d 130000 |d 130700 |e 130000PH1068900059 |e 130700PH1068900059 |k 0/130000/ |k 1/130000/130700/ |p 5 | ||
| 998 | |g 1076034802 |a Krufczik, Matthias |m 1076034802:Krufczik, Matthias |d 130000 |d 130700 |e 130000PK1076034802 |e 130700PK1076034802 |k 0/130000/ |k 1/130000/130700/ |p 1 |x j | ||
| 999 | |a KXP-PPN1560088648 |e 2972303415 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"note":["Gesehen am 22.06.2017"],"title":[{"title_sort":"Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structure","title":"Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structure"}],"id":{"eki":["1560088648"],"doi":["10.3390/ijms18051005"]},"recId":"1560088648","language":["eng"],"origin":[{"dateIssuedKey":"2017","dateIssuedDisp":"7 May 2017"}],"type":{"media":"Online-Ressource","bibl":"article-journal"},"relHost":[{"type":{"media":"Online-Ressource","bibl":"periodical"},"origin":[{"dateIssuedDisp":"2000-","publisherPlace":"Basel","dateIssuedKey":"2000","publisher":"Molecular Diversity Preservation International"}],"language":["eng"],"physDesc":[{"extent":"Online-Ressource"}],"part":{"extent":"14","year":"2017","text":"18(2017,5) Artikel-Nummer 1005, 16 Seiten","issue":"5","volume":"18"},"title":[{"title":"International journal of molecular sciences","title_sort":"International journal of molecular sciences"}],"id":{"zdb":["2019364-6"],"eki":["316340715"],"issn":["1422-0067","1661-6596"]},"disp":"Combining low temperature fluorescence DNA-hybridization, immunostaining, and super-resolution localization microscopy for nano-structure analysis of ALU elements and their influence on chromatin structureInternational journal of molecular sciences","note":["Gesehen am 17.09.20"],"recId":"316340715","titleAlt":[{"title":"IJMS"}],"pubHistory":["1.2000 -"]}],"name":{"displayForm":["Matthias Krufczik, Aaron Sievers, Annkathrin Hausmann, Jin-Ho Lee, Georg Hildenbrand, Wladimir Schaufler and Michael Hausmann"]},"person":[{"given":"Matthias","display":"Krufczik, Matthias","role":"aut","family":"Krufczik","roleDisplay":"VerfasserIn"},{"roleDisplay":"VerfasserIn","family":"Hildenbrand","given":"Georg Lars","display":"Hildenbrand, Georg Lars","role":"aut"},{"given":"Michael","display":"Hausmann, Michael","role":"aut","roleDisplay":"VerfasserIn","family":"Hausmann"}],"physDesc":[{"extent":"14 S."}]} | ||
| SRT | |a KRUFCZIKMACOMBININGL7201 | ||