ELI-Beamlines (ELI-BL), one of the three pillars of the Extreme Light Infrastructure endeavour, will be in a unique position to perform research in high-energy-density-physics (HEDP), plasma physics and ultra-high intensity (UHI) (1022W/cm2) laser–plasma interaction. Recently the need for HED laboratory physics was identified and the P3 (plasma physics platform) installation under construction in ELI-BL will be an answer. The ELI-BL 10 PW laser makes possible fundamental research topics from high-field physics to new extreme states of matter such as radiation-dominated ones, high-pressure quantum ones, warm dense matter (WDM) and ultra-relativistic plasmas. HEDP is of fundamental importance for research in the field of laboratory astrophysics and inertial confinement fusion (ICF). Reaching such extreme states of matter now and in the future will depend on the use of plasma optics for amplifying and focusing laser pulses. This article will present the relevant technological infrastructure being built in ELI-BL for HEDP and UHI, and gives a brief overview of some research under way in the field of UHI, laboratory astrophysics, ICF, WDM, and plasma optics.

SummaryFluorescence resonance energy transfer (FRET) by acceptor photobleaching is a simple but effective tool for measurements of protein-protein interactions. Until recently, it has been restricted to qualitative or relative assessments owing to the spectral bleed-through contamination resulting from fluorescence overlap between the donor and the acceptor. In this paper, we report a quantitative algorithm that combines the spectral unmixing technique with FRET by acceptor photobleaching. By spectrally unmixing the emissions before and after photobleaching, it is possible to resolve the spectral bleed-through and retrieve the FRET efficiency/interaction distance quantitatively. Using a human keratinocyte cell line transfected with cyan fluorescent protein (CFP)-and yellow fluorescent protein (YFP)-tagged Cx26 connexins as an example, FRET information at homotypic gap junctions is measured and compared with well-established methods. Results indicate that the new approach is sensitive, flexible, instrument independent and solely FRET dependent. It can achieve FRET estimations similar to that from a sensitized emission FRET method. This approach has a great advantage in providing the relative concentrations of the donor and the acceptor; this is, for example, very important in the comparative study of cell populations with variable expression levels.

Multiple connexins, the major proteins of gap junctions, have overlapping expression in the human epidermis and are postulated to have a key role in keratinocyte differentiation and homeostasis. The functional importance of connexins in the epidermis is emphasised by the association of mutations in four human connexins with various hyperproliferative skin disorders. As immunohistochemistry demonstrated overlapping expression of specific connexins in keratinocytes, we performed colocalisation analyses and applied a modified FRET methodology to assess possible heteromeric interactions between different combinations of four wild-type (wt) and mutant connexins. The data generated indicate that there is evidence for multiple connexin interactions at the plasma membrane between (wt)Cx26, (wt)Cx30 and (wt)Cx31 in keratinocytes and thus, the potential for the formation of a large number of different channel types each with different channel properties. In addition, we demonstrate that the inherent in vitro trafficking defect of the skin disease mutations (D50N)Cx26 and (G11R)Cx30 can be overcome partially by the coexpression of different wild-type connexins but this rescue does not result in large gap junction aggregates at the plasma membrane. These data indicate that skin disease associated Cx26 or Cx30 mutations are likely to disrupt a number of different channel types important in distinct aspects of keratinocyte biology.