Tracking the molecular signature of developing skeletal tissues.

We isolated cells from their native in vivo microenvironment using the Laser Capture Micro dissection (LCM). Bone and cartilage tissues were studied from mouse embryonic (18dpc) processed by cry sections enabled the cell isolation from anatomical complexity of skeletal tissues using the LCM technique. RNA was purified from the isolated cells and followed with amplification stage to hybridize on gene array for high through (HT) put analysis to profile the tissues gene expression.

Dielectric dispersion of suspended cells using 3D reconstructed morphology model.

In the framework of this study, novel method for dispersion analysis of cellular suspensions is presented. The method is fundamentally based on the ability to reconstruct the exact 3D morphology of a given cell with resolution accuracy of few nanometers using AFM imaging. By applying a reverse engineering approach, the morphology of the cell is constructed based on a set of measured spatial points that describes its geometry.

Application of the laser capture microdissection technique for molecular definition of skeletal cell differentiation in vivo.

Laser capture microdissection (LCM) method allows selection of individual or clustered cells from intact tissues. This technology enables one to pick cells from tissues that are difficult to study individually, sort the anatomical complexity of these tissues, and make the cells available for molecular analyses. Following the cells' extraction, the nucleic acids and proteins can be isolated and used for multiple applications that provide an opportunity to uncover the molecular control of cellular fate in the natural microenvironment.

Cell-based screening for membranal and cytoplasmatic markers using dielectric spectroscopy.

Dielectric spectroscopy (DS) of living biological cells is based on the analysis of the complex dielectric permittivity of cells suspended in a physiological medium. It provides knowledge on the polarization-relaxation response of cells to external electric field as function of the excitation frequency. This response is strongly affected by both structural and molecular properties of cells and therefore, can reveal rare insights on cell physiology and behaviour.

Site localization of membrane-bound proteins on whole cell level using atomic force microscopy.

This study presents molecular recognition method, which is based on specific force measurements between modified AFM (atomic force microscopy) tip and mammalian cell. The presented method allows recognition of specific cell surface proteins and receptor sites by nanometer accuracy level. Here we demonstrate specific recognition of membrane-bound Osteopontin (OPN) sites on preosteogenic cell membrane.

Laser capture microdissection and laser pressure catapulting as tools to study gene expression in individual cells of a complex tissue.

Laser capture microdissection (LCM) method allows the selection of individual or clustered cells from intact tissues. LCM enables to pick cells from tissues that are difficult to study individually, to sort the anatomical complexity of tissues, and to make the cells available for molecular analyses. This technology provides an opportunity to uncover the molecular control of cellular fate in the natural microenvironment.

Dexamethasone regulation of cFos mRNA in osteoprogenitors.

The glucocorticoid, dexamethasone (Dex), has a beneficial effect on osteogenesis by increasing the activation and differentiation of osteoblastic cells. We investigated the effect of Dex on osteoblasts and detected changes in pattern of expression of cFos mRNA. cFos is an early response gene that is expressed as unspliced and spliced mRNAs.