Linked Life Data

10670771

Source:http://linkedlifedata.com/resource/pubmed/id/10670771

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
2000-3-2
pubmed:abstractText
The intracellular pH (pHin) of Saccharomyces cerevisiae was measured employing fluorescence ratio imaging microscopy (FRIM). The yeast cells were fluorescently labeled with the pH dependent probe 5(and-6)-carboxyfluorescein (cF) or 5(and-6)-carboxyfluorescein succinimidyl ester (cFSE), and subsequently attached to ferric nitrate pretreated glass slides. The labeled and adhered cells could still divide and were metabolically active. Measurement of the pHin was performed during continuous perfusion of the cells with buffer or medium. Cells labeled with cF are highly fluorescent and in non-energized cells the pHin could be easily measured. However, in energized yeast cells cF was accumulated in the vacuoles and/or exported to the extracellular environment, most likely by an energy-dependent transport system, thus limiting the time period over which the pHin can be effectively measured. Therefore, cFSE (which conjugates with aliphatic amines in the cytoplasm) was applied to prevent translocation of fluorescent probe to the vacuole and/or extracellular environment. The continuous perfusion in combination with the cFSE labeling of the immobilized cells was successfully applied to determine the effect of low and high pHin and addition of glucose on the pHin of individual yeast cells over a long time period.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0167-7012
pubmed:author
pubmed:issnType
Print
pubmed:volume
39
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
253-64
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
2000
pubmed:articleTitle
Assessment of the intracellular pH of immobilized and continuously perfused yeast cells employing fluorescence ratio imaging analysis.
pubmed:affiliation
Department of Food Technology and Nutritional Sciences, Wageningen University and Research Center, The Netherlands. pieter.breeuwer@micro.fdsci.wau.nl
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't