FLIPPER, a combinatorial probe for correlated live imaging and electron microscopy, allows identification and quantitative analysis of various cells and organelles.

Ultrastructural examination of cells and tissues by electron microscopy (EM) yields detailed information on subcellular structures. However, EM is typically restricted to small fields of view at high magnification; this makes quantifying events in multiple large-area sample sections extremely difficult. Even when combining light microscopy (LM) with EM (correlated LM and EM: CLEM) to find areas of interest, the labeling of molecules is still a challenge.

Prolonged cigarette smoke exposure alters mitochondrial structure and function in airway epithelial cells.

Background: Cigarette smoking is the major risk factor for COPD, leading to chronic airway inflammation. We hypothesized that cigarette smoke induces structural and functional changes of airway epithelial mitochondria, with important implications for lung inflammation and COPD pathogenesis.

Methods: We studied changes in mitochondrial morphology and in expression of markers for mitochondrial capacity, damage/biogenesis and fission/fusion in the human bronchial epithelial cell line BEAS-2B upon 6-months from ex-smoking COPD GOLD stage IV patients to age-matched smoking and never-smoking controls.

Destruction of tissue, cells and organelles in type 1 diabetic rats presented at macromolecular resolution.

Finding alternatives for insulin therapy and making advances in etiology of type 1 diabetes benefits from a full structural and functional insight into Islets of Langerhans. Electron microscopy (EM) can visualize Islet morphology at the highest possible resolution, however, conventional EM only provides biased snapshots and lacks context. We developed and employed large scale EM and compiled a resource of complete cross sections of rat Islets during immuno-destruction to provide unbiased structural insight of thousands of cells at macromolecular resolution.

Immunolabeling artifacts and the need for live-cell imaging.

Fluorescent fusion proteins have revolutionized examination of proteins in living cells. Still, studies using these proteins are met with criticism because proteins are modified and ectopically expressed, in contrast to immunofluorescence studies. However, introducing immunoreagents inside cells can cause protein extraction or relocalization, not reflecting the in vivo situation.

Stwl modifies chromatin compaction and is required to maintain DNA integrity in the presence of perturbed DNA replication.

Hydroxyurea, a well-known DNA replication inhibitor, induces cell cycle arrest and intact checkpoint functions are required to survive DNA replication stress induced by this genotoxic agent. Perturbed DNA synthesis also results in elevated levels of DNA damage. It is unclear how organisms prevent accumulation of this type of DNA damage that coincides with hampered DNA synthesis.

Cerebellar heterokaryon formation increases with age and after irradiation.

Hematopoietic cells have been demonstrated to survive in many nonhematopoietic tissues after transplantation. Apparent "bone marrow-derived" cerebellar Purkinje cells in fact result from fusion events and it has been suggested that fusion may be a natural physiological phenomenon to rescue dysfunctioning cells. Here, we show that fusion of transplanted bone marrow cells with resident Purkinje cells is age-dependent and is strongly enhanced when Purkinje cells are damaged by high-dose irradiation.

Polarised asymmetric inheritance of accumulated protein damage in higher eukaryotes.

Disease-associated misfolded proteins or proteins damaged due to cellular stress are generally disposed via the cellular protein quality-control system. However, under saturating conditions, misfolded proteins will aggregate. In higher eukaryotes, these aggregates can be transported to accumulate in aggresomes at the microtubule organizing center.

Early hypergravity exposure effects calbindin-D28k and inositol-3-phosphate expression in Purkinje cells.

In this study the effects of hypergravity were analyzed on cerebellar Purkinje cells during early development in rats. The cerebellum is a key structure in the control and the adaptation of posture and anti-gravity activities. This holds particularly when external conditions are modified.

Functional and anatomic alterations in the gentamicin-damaged vestibular system in the guinea pig.

Hypothesis: The purpose of this study was to investigate the expected functional and morphologic effect of gentamicin on the vestibular system simultaneously by measurement of vestibular evoked potentials and electron microscopic evaluation.

Background: Vestibular short-latency evoked potentials to linear acceleration have been shown to be a useful parameter of vestibular function. In gentamicin-treated animals, the morphologic damage has been well documented, although this has seldom been quantified.