Identification of single fluorescent labels using spectroscopic microscopy

Applied Spectroscopy
Emily C HeiderJoel M Harris

Abstract

Detection of single, fluorescently labeled biomolecules is providing a powerful approach to measuring molecular transport, biomolecular interactions, and localization in biological systems. Because the biological molecules of interest rarely exhibit sufficient intrinsic fluorescence to allow observation of individual molecules, they are usually labeled with fluorescent dye molecules, fluorescent proteins, semiconductor nanocrystals or quantum dots, or fluorescently doped silica or polymer nanospheres to allow their detection. Differences in the photophysical and spectral properties of different labels allow one to identify individual molecules by distinguishing their corresponding labels. A simple approach to measuring fluorescence spectra of individual fluorescent labels can be implemented in a standard wide-field fluorescence microscope, where a grating or prism is incorporated into the path from the microscope to an imaging detector to disperse the emission spectrum. In this work, principal components and cluster analysis are applied to the identification of fluorescence spectra from single fluorescent labels, with statistical tests of the classification results. Spectra are determined from diffracted images of fluorescent n...Continue Reading

References

Sep 25, 1998·Science·M BruchezA P Alivisatos
Oct 6, 1998·Annual Review of Biochemistry·R Y Tsien
Apr 1, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·J HartstraR A de Zeeuw
Aug 10, 2000·Proceedings of the National Academy of Sciences of the United States of America·T D LacosteS Weiss
Oct 12, 2000·Analytical Chemistry·Y MaE S Yeung
Mar 8, 2003·Methods in Enzymology·James E N JonkmanErnst H K Stelzer
Apr 10, 2004·Optics Letters·Gerhard A BlabThomas Schmidt
Aug 19, 2007·Behavior Research Methods·Otto H MacLin, Colin M Phelan
Apr 9, 2008·Biophysical Journal·Jody L Swift, David T Cramb
Dec 25, 2008·Chemical Reviews·M Sameiro T Gonçalves
Jan 2, 2009·Analytical Chemistry·Joshua R Wayment, Joel M Harris

❮ Previous
Next ❯

Citations

Oct 22, 2016·Analytical and Bioanalytical Chemistry·Emily C HeiderAndres D Campiglia
Dec 10, 2015·Applied Spectroscopy·Nirvani MujumdarAndres D Campiglia
Oct 4, 2011·Analytical Chemistry·Emily C HeiderJoel M Harris
May 18, 2011·Analytical Chemistry·Emily C HeiderJoel M Harris

❮ 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.