Flavin fluorescence lifetime imaging of living peripheral blood mononuclear cells on micro and nano-structured surfaces

International audienceFabricated micro-and nano-structured surfaces were evaluated for use with living cells. Metabolic state was tested by means of endogenous flavin fluorescence of living peripheral blood mononuclear cells (PBMC) positioned on a coverslip, non-covered, or covered with micro-or nano-structured surfaces (OrmoComp polymer structures produced by 2-photon photopolymerisation, or Zinc Oxide (ZnO) layer fabricated by pulsed laser deposition). Confocal microscopy and Fluorescence Lifetime Imaging Microscopy (FLIM) were employed to gather flavin fluorescence lifetime images of living PBMC on structured surfaces. Gathered data are the first step towards monitoring of the live cell interaction with different micro/nano-structured surfaces and thus evaluate their potential applicability in the biomedical field

Estimation of area at risk in myocardial infarction

This study presents a new method for estimation and imaging of the area at risk (AaR) in myocardial infarction (MI). The values of the ST-segment deviations of 12-lead ECG signal were used as input parameters. Based on DECARTO model, the spherical surface was chosen as a reference surface to approximate the ventricular wall. On this surface, the spatial ST vector was projected. The center of AaR was defined as an intersection of the spatial ST vector with spherical surface; the size of the AaR was set to be proportional to the number of electrical leads with ST- segment deviations. The method was tested using data of 10 patients with acute MI. The visual comparison showed good agreement with the AaRECG estimates based originally on the Selvester QRS scoring as well as with a non- electrocardiographic imaging method (SPECT)

Characterization of the Interaction of Hypericin with Protein Kinase C in U-87 MG Human Glioma Cells

A fluorescence imaging technique was used to monitor intracellular localization of protein kinase C (PKC) in U-87 MG human glioma cells in the presence of hypericin (Hyp) and phorbol 12-myristate-13-acetate (PMA). It is shown that PKC localization, which reflects its activity, is influenced by Hyp and this influence is different from that observed for PMA which acts as PKC activator. Fluorescence binding experiments were used to determine the binding constants of Hyp to several isoforms of PKC. The obtained values of K(d)s (approximately 100 nM) suggest that Hyp binds with high affinity to PKC. Finally, molecular modeling was used to compare structural models of the interaction of C1B domain of PKC (PKC isoforms alpha, delta, gamma) with Hyp and our previously published model of the (C1B domain PKCgamma)/PMA complex. The influence of Hyp on PKC translocation in U-87 MG cells in comparison with PMA, colocalization fluorescence pattern of Hyp and PKC, the higher binding affinity of Hyp to PKC in comparison with known binding constants of phorbol esters, as well as the binding mode of Hyp and PMA to the C1B domain of PKC suggested by molecular modeling, support the idea that Hyp and PMA might competitively bind to the regulatory domain of PKC