Response to "Comments on the Fluorescein Excitation and Emission Polarization Spectra in Living Cells"

Cercek, L; Cercek, Bojan

doi:10.1016/s0006-3495(79)85185-1

Biophysical Journal

1979

DOI: 10.1016/s0006-3495(79)85185-1

|View full text |Cite

Response to "Comments on the Fluorescein Excitation and Emission Polarization Spectra in Living Cells"

L Cercek¹,

Bojan Cercek²

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...

Citation Types

Supporting

Mentioning

Contrasting

Year Published

1981

Publication Types

Select...

Article1

Relationship

Self Cite0

Independent1

Authors

Journals

Cited by 1 publication

(1 citation statement)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Udkoff and Norman (21) concluded that the intracellular fluorescein was partly bound to proteins. Cercek et al ( 4 ) argued that pH, polarity and viscosity vary between different microdomains in the cytoplasm, giving rise to several different solvation states of fluorescein with varying fluorescence characteristics (4). Their application of fluorochromasia in a cancer test (3) has given the nature of intracellular fluorescein binding new importance.…”

mentioning

confidence: 99%

Intracellular binding of fluorescein in lymphocytes

Meisingset

Steen

1981

Cytometry

View full text Add to dashboard Cite

The fluorescence characteristics of intracellular fluorescein, formed by hydrolysis of fluorescein diacetate in peripheral human lymphocytes, were studied by fluorometry on cell suspensions and compared to those of albumin bound and free fluorescein in solution. The absorption and fluorescence spectra of both intracellular fluorescein and fluorescein in aqueous solutions of albumin and glycerol were red shifted by 2-10 nm as compared to the spectra of fluorescein in phosphate buffered saline. The fluorescence polarization (P) of both intracellular fluorescein and a mixture of albumin-bound and free fluorescein showed a decrease towards low emission wavelengths and an increase toward high excitation wavelengths. The results were found to be consistent with a simple model assuming that part of the intracellular fluorescein is dissolved in the aqueous phase of the cytoplasm, giving P < 0.1, while the rest is bound to macromolecules, giving P = 0.33. The fraction of bound intracellular fluorescein was estimated to be about 70%. Fluorescein was found to bind with higher affinity and more rigidly (P = 0.43) to albumin than to intracellular macromolecules in general. In order to elucidate the state of intracellular fluorescein further, we have studied the fluorescence characteristics of intracellular fluorescein and compared them to those of fluorescein in phosphate buffered saline (PBS) and in solutions of glycerol and albumin in PBS, glycerol serving as a medium of lower polarity than water and albumin as a model for fluorescein binding proteins. The results are consistent with the assumption that part of the intracellular fluorescein is bound to macromolecules while the rest is dissolved in intracellular water. TheoryTo make a simple model for intracellular fluorescein binding, we assume that the fluorescence from the cells during FDA hydrolysis consists of two components: ( a ) fluorescence from unbound fluores-272

show abstract

mentioning

confidence: 99%

Intracellular binding of fluorescein in lymphocytes

Meisingset

Steen

1981

Cytometry

View full text Add to dashboard Cite

show abstract

scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.

Contact Info

customersupport@researchsolutions.com

10624 S. Eastern Ave., Ste. A-614

Henderson, NV 89052, USA

This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.

Blog Terms and Conditions API Terms Privacy Policy Contact Cookie Preferences Do Not Sell or Share My Personal Information

Made with 💙 for researchers

Part of the Research Solutions Family.

Response to "Comments on the Fluorescein Excitation and Emission Polarization Spectra in Living Cells"

Cited by 1 publication

References 9 publications

Intracellular binding of fluorescein in lymphocytes

Intracellular binding of fluorescein in lymphocytes

Contact Info

Product

Resources

About