Tutorial: Measurement of fluorescence spectra and determination of relative fluorescence quantum yields of transparent samples

Methods and Applications in Fluorescence
Marcia Levitus

Abstract

The measurement of fluorescence spectra and the determination of fluorescence quantum yields in transparent samples are conceptually simple tasks, but these procedures are subject to several pitfalls that can lead to significant errors. Available technical reports and protocols often assume that the reader possesses a solid theoretical background in spectroscopy and has ample experience with fluorescence instrumentation, but this is often not the case given the many applications of fluorescence in diverse fields of science. The goal of this tutorial is to provide a didactic treatment of the topic that will hopefully be accessible to readers without extensive expertise in the field of fluorescence. The article covers the theoretical background needed to understand the origins of the most common artifacts researchers can expect. Possible artifacts are illustrated with examples to help readers avoid them or identify them if present. A step-by-step example of a fluorescence quantum yield determination in solution is provided with detailed experimental information to help readers understand how to design and analyze experiments.

References

Jan 1, 1978·Annual Review of Biochemistry·L Stryer
Apr 7, 2007·The Review of Scientific Instruments·Paul C DeRoseGary W Kramer
Aug 28, 2007·The Journal of Physical Chemistry. B·Matthew E SanbornMarcia Levitus
Jan 6, 2009·Nursing·Cara M HudsonCathy C Harro
Oct 16, 2010·Photochemistry and Photobiology·R Bryan SearsClaudia Turro
Jan 22, 2011·Analytical Chemistry·Knut Rurack, Monika Spieles
Jul 30, 2011·Chemical Society Reviews·Yuning HongBen Zhong Tang
Jul 23, 2013·Nature Protocols·Christian WürthUte Resch-Genger
Oct 22, 2013·Chemical Society Reviews·Elana M S StennettMarcia Levitus
Jan 1, 2008·Journal of Research of the National Institute of Standards and Technology·Adolfas K Gaigalas, Lili Wang
Feb 10, 2018·Methods and Applications in Fluorescence·J VangindertaelK P F Janssen
Mar 26, 2019·Analytical Chemistry·Krzysztof Nawara, Jacek Waluk
Sep 26, 2019·Methods and Applications in Fluorescence·Shivaji N TayadeKiran Kumar K Sharma

❮ Previous
Next ❯

Citations

Nov 26, 2020·Methods and Applications in Fluorescence·David J S BirchYves Mely
Mar 18, 2021·Journal of the American Society for Mass Spectrometry·Eliise TammekiviIvo Leito

❮ 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.