PMID: 11933012Apr 5, 2002Paper

Discrimination of DNA and RNA in cells by a vital fluorescent probe: lifetime imaging of SYTO13 in healthy and apoptotic cells

Cytometry
Marc A M J van ZandvoortDick W Slaaf

Abstract

Of the few vital DNA and RNA probes, the SYTO dyes are the most specific for nucleic acids. However, they show no spectral contrast upon DNA or RNA binding. We show that fluorescence lifetime imaging using two-photon excitation of SYTO13 allows differential and simultaneous imaging of DNA and RNA in living cells, as well as sequential and repetitive assessment of staining patterns. Two-photon imaging of SYTO13 is combined with lifetime contrast, using time-gated detection. We focus on distinguishing DNA and RNA in healthy and apoptotic Chinese hamster ovary cells. In healthy cells, SYTO13 has a fluorescence lifetime of 3.4 +/- 0.2 ns when associated with nuclear DNA. Bound to RNA, its lifetime is 4.1 +/- 0.1 ns. After induction of apoptosis, clusters of SYTO13 with fluorescence lifetime of 3.4 +/- 0.2 ns become apparent in the cytoplasm. They are identified as mitochondrial DNA on the basis of colocalization experiments with the DNA-specific dye, DRAQ5, and the mitochondrial-specific dye, CMXRos. Upon progression of apoptosis, the lifetime of SYTO13 attached to DNA shortens significantly, which is indicative of changes in the molecular environment of the dye. We have characterized SYTO13 as a vital lifetime probe, allowing repe...Continue Reading

References

Nov 13, 1992·Cell·T N Davis
May 1, 1992·Analytical Biochemistry·J R LakowiczM Johnson
Mar 1, 1992·Cell Calcium·J R LakowiczM L Johnson
Apr 6, 1990·Science·W DenkW W Webb
Aug 1, 1994·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·R M WilliamsW W Webb
May 1, 1997·Experimental Cell Research·M YoshidaS Horinouchi
Mar 21, 1998·Methods in Cell Biology·T FrenchE Gratton
Jun 26, 1998·Current Opinion in Cell Biology·T Misteli, D L Spector
Dec 16, 1998·The Journal of Cell Biology·H MaR Berezney
Mar 30, 1999·Analytical Biochemistry·H J LinJ R Lakowicz
Jun 25, 1999·Family Practice·W J de GrauwC van Weel
Oct 26, 1999·Current Biology : CB·F S Wouters, P I Bastiaens
Jan 29, 2000·Critical Reviews in Eukaryotic Gene Expression·H LeonhardtM C Cardoso
Jan 29, 2000·Journal of Microscopy·A SquireP I Bastiaens
Feb 24, 2000·The Journal of Pathology·M M Kockx, M W Knaapen
Mar 29, 2000·Biophysical Journal·P J VerveerP I Bastiaens
Mar 29, 2000·Biophysical Journal·G H Patterson, D W Piston
Jun 27, 2000·Journal of Molecular Recognition : JMR·O KirschsteinL Kittler
Jun 28, 2000·Analytical Biochemistry·S DespaM Ameloot
Oct 19, 2000·Experimental Cell Research·H D HalickaZ Darzynkiewicz
Nov 23, 2000·European Journal of Cell Biology·M StraubS W Hell
Nov 23, 2000·Genes & Development·B K KennedyE Harlow
Dec 7, 2000·Journal of Microscopy·K König
Oct 1, 1995·Sensors and Actuators. B, Chemical·Henryk Szmacinski, Joseph R Lakowicz

❮ Previous
Next ❯

Citations

Oct 9, 2002·Current Opinion in Neurobiology·Fritjof Helmchen, Winfried Denk
Nov 6, 2007·Applied and Environmental Microbiology·Petr WalczyskoThomas R Neu
May 26, 2004·BMC Biology·Francisco J IborraPeter R Cook
Feb 1, 2014·Reproductive Medicine and Biology·Katsumi KasashimaHitoshi Endo
Oct 14, 2006·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Tomohiro UchimuraTotaro Imasaka
Dec 17, 2003·Microscopy Research and Technique·W BeckerC Biskup
Dec 21, 2005·Applied Optics·Kai HasslerTheo Lasser
Aug 6, 2005·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Robert M MartinM Cristina Cardoso
Feb 22, 2003·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·H Helen CuiHarry A Crissman
Mar 30, 2007·Microscopy Research and Technique·Wolfgang BeckerChristoph Biskup
Mar 17, 2007·Microscopy Research and Technique·A RückR Steiner
Feb 9, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Donald WlodkowicZbigniew Darzynkiewicz
Jun 6, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Iva SlaninováEva Táborská
Sep 14, 2006·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Kerenza L NjohPaul J Smith
Jan 4, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Donald WlodkowicJukka Pelkonen
May 25, 2012·Journal of Microscopy·W Becker
Sep 22, 2011·Journal of Microscopy·F FereidouniH C Gerritsen
Jan 22, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·G Warnes, S Martins
Mar 29, 2014·Electrophoresis·Wenyan LiJessica P Houston
Jan 28, 2010·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Anirudh J UllalCharles F Reich
Sep 17, 2005·Journal of Immunological Methods·Rosemary L Sparrow, Emma Tippett
Apr 4, 2012·FEMS Immunology and Medical Microbiology·Alexandra S PennJeremy S Webb
Feb 26, 2010·FEMS Microbiology Ecology·Thomas R NeuJohn R Lawrence
Jan 25, 2017·Lab on a Chip·Andrew C HodgsonKevin J Chalut
Jan 1, 2011·Pharmaceutics·Beverley IsherwoodNeil O Carragher
Jan 18, 2005·Physiology·Grace E Stutzmann, Ian Parker
Mar 27, 2009·American Journal of Physiology. Cell Physiology·R X FariaL A Alves
Sep 5, 2008·Current Protocols in Cytometry·Donald Wlodkowic, Joanna Skommer
Jun 18, 2015·Methods and Applications in Fluorescence·Satoshi KawanabeTotaro Imasaka
Aug 29, 2018·Journal of X-ray Science and Technology·Kunio ShinoharaShigenobu Toné
Apr 21, 2018·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Lei WangJinqing Qu
Sep 5, 2008·Current Protocols in Cytometry·Paul J SmithRachel J Errington
Apr 14, 2021·Langmuir : the ACS Journal of Surfaces and Colloids·Aloke BapliDebabrata Seth

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

© 2022 Meta ULC. All rights reserved