Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients

The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins...

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Autores principales:	Ingrid Lajdova, Livia Ovsonkova, Viera Spustova, Adrian Oksa, Dusan Chorvat, Anton Mateasik, Alzbeta Marcek Chorvatova
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	22-NBD-cholesterol CKD—chronic kidney disease FLIM—fluorescence lifetime imaging microscopy PBMC—peripheral blood mononuclear cells spectrally resolved confocal microscopy statins Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/6fd3c0554fec4552b56d530de68fb990
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spelling	oai:doaj.org-article:6fd3c0554fec4552b56d530de68fb9902021-11-25T18:27:20ZSpectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients10.3390/molecules262268001420-3049https://doaj.org/article/6fd3c0554fec4552b56d530de68fb9902021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6800https://doaj.org/toc/1420-3049The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins. Blood samples from healthy volunteers (HV) and CKD patients, either treated or untreated with statins, were compared. Spectral imaging was done using confocal microscopy at 16 spectral channels in response to 458 nm excitation. Time-resolved imaging was achieved by time-correlated single photon counting (TCSPC) following excitation at 475 nm. The fluorescence of 22-NBD-cholesterol was mostly integrated into plasmatic membrane and/or intracellular membrane but was missing from the nuclear region. The presence of two distinct spectral forms of 22-NBD-cholesterol was uncovered, with significant variations between studied groups. In addition, two fluorescence lifetime components were unmasked, changing in CKD patients treated with statins. The gathered results indicate that 22-NBD-cholesterol may serve as a tool to study changes in the lipid metabolism of patients with CKD to monitor the effect of statin treatment.Ingrid LajdovaLivia OvsonkovaViera SpustovaAdrian OksaDusan ChorvatAnton MateasikAlzbeta Marcek ChorvatovaMDPI AGarticle22-NBD-cholesterolCKD—chronic kidney diseaseFLIM—fluorescence lifetime imaging microscopyPBMC—peripheral blood mononuclear cellsspectrally resolved confocal microscopystatinsOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6800, p 6800 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	22-NBD-cholesterol CKD—chronic kidney disease FLIM—fluorescence lifetime imaging microscopy PBMC—peripheral blood mononuclear cells spectrally resolved confocal microscopy statins Organic chemistry QD241-441
spellingShingle	22-NBD-cholesterol CKD—chronic kidney disease FLIM—fluorescence lifetime imaging microscopy PBMC—peripheral blood mononuclear cells spectrally resolved confocal microscopy statins Organic chemistry QD241-441 Ingrid Lajdova Livia Ovsonkova Viera Spustova Adrian Oksa Dusan Chorvat Anton Mateasik Alzbeta Marcek Chorvatova Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
description	The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins. Blood samples from healthy volunteers (HV) and CKD patients, either treated or untreated with statins, were compared. Spectral imaging was done using confocal microscopy at 16 spectral channels in response to 458 nm excitation. Time-resolved imaging was achieved by time-correlated single photon counting (TCSPC) following excitation at 475 nm. The fluorescence of 22-NBD-cholesterol was mostly integrated into plasmatic membrane and/or intracellular membrane but was missing from the nuclear region. The presence of two distinct spectral forms of 22-NBD-cholesterol was uncovered, with significant variations between studied groups. In addition, two fluorescence lifetime components were unmasked, changing in CKD patients treated with statins. The gathered results indicate that 22-NBD-cholesterol may serve as a tool to study changes in the lipid metabolism of patients with CKD to monitor the effect of statin treatment.
format	article
author	Ingrid Lajdova Livia Ovsonkova Viera Spustova Adrian Oksa Dusan Chorvat Anton Mateasik Alzbeta Marcek Chorvatova
author_facet	Ingrid Lajdova Livia Ovsonkova Viera Spustova Adrian Oksa Dusan Chorvat Anton Mateasik Alzbeta Marcek Chorvatova
author_sort	Ingrid Lajdova
title	Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
title_short	Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
title_full	Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
title_fullStr	Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
title_full_unstemmed	Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
title_sort	spectrally and time-resolved fluorescence imaging of 22-nbd-cholesterol in human peripheral blood mononuclear cells in chronic kidney disease patients
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/6fd3c0554fec4552b56d530de68fb990
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Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients

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