Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture. To avoid tedious and often painful manual dissociations the gentleMACS Dissociator allows one to dissociate tissue very efficiently under controlled and reproducible conditions. The gentleMACS Dissociator can optimally dissociate mouse spleen, combining timesaving and standardization with user-safety. This video describes how to dissociate mouse spleens using the gentleMACS Dissociator, an automated bench-top device that can mechanically disrupt tissues using special tubes to produce viable cell suspensions. Following dissociation, spleens are filtered, centrifuged, and resuspended for further applications

Generation of Single-Cell Suspensions from Mouse Neural Tissue

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a repertoire of cell surface molecules, or molecular determinants, through which it can be identified and characterized. Currently, robust cell identification and separation technologies require single-cell preparations to be generated while simultaneously limiting cell death and destruction of characteristic surface protein. The gentleMACS Dissociator, when used in combination with trypsin or papain-based dissociation kits, can effectively and gently dissociate brain tissue while preserving antigen epitopes and limiting cell loss. Standardized preparation of single-cell suspensions is achieved using C Tubes and optimized, preset gentleMACS Programs. Once generated, single-cell suspensions can be treated with monoclonal conjugates like Anti-Prominin-1 MicroBeads, which identify neural progenitors, or purified further using Myelin Removal Beads

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of dissociating whole organs requires specific parameters in order to obtain a high number of viable cells in a reproducible manner. The gentleMACS Dissociator optimizes this task with a simple, practical protocol. The instrument contains pre-programmed settings that are optimized for the efficient but gentle dissociation of a variety of tissue types, including mouse lungs. In this publication the use of the gentleMACS Dissociator on lung tissue derived from mice is demonstrated

Strukturiertes Komposit aus Matrixmaterial und Nanopartikeln

The invention relates to a composite, which can be produced by photostructuring a photostructurable matrix material in a composite formulation, a structured matrix with nanoparticles thus being formed, wherein the index of refraction of the composite with nanoparticles differs from the index of refraction of the composite without nanoparticles at a wavelength selected from the range of 150 nm to 2000 nm by less than 0.5, the composite being hierarchically structured and comprising at least one structural unit (I) having a selected thickness (i) and structural units (II) deviating from the structural unit (I) and having a selected thickness (ii), the thickness (ii) at the branch-off points being at most half of the thickness (i). The invention further relates to an improved method for producing a composite that comprises photostructured matrix material and nanoparticles contained therein, and to the use of the composite

Freeform direct laser writing of versatile topological 3D scaffolds enabled by intrinsic support hydrogel

In this study, a novel approach to create arbitrarily shaped 3D hydrogel objects is presented, wherein freeform two-photon polymerization (2PP) is enabled by the combination of a photosensitive hydrogel and an intrinsic support matrix. This way, topologies without physical contact such as a highly porous 3D network of concatenated rings were realized, which are impossible to manufacture with most current 3D printing technologies. Micro-Raman and nanoindentation measurements show the possibility to control water uptake and hence tailor the Young's modulus of the structures via the light dosage, proving the versatility of the concept regarding many scaffold characteristics that makes it well suited for cell specific cell culture as demonstrated by cultivation of human induced pluripotent stem cell derived cardiomyocytes.Peer reviewe

Case Management zur Erhaltung von Arbeits- und Ausbildungsverhaeltnissen behinderter Menschen (CMB) Zweiter Sachstandsbericht der wissenschaftlichen Begleitung einer Modellinitiative der Bundesarbeitsgemeinschaft fuer Rehabilitation

Der Forschungsbericht thematisiert zunaechst die Arbeit der Begleitforschung im zweiten Projektjahr und den Stand des CMB-Modellprojekts. Dazu gehoeren zum einen die Aktivitaeten der Begleitforschung im Berichtszeitraum, wie die Bearbeitung von TeilnehmerInnendaten, Arbeitssitzungen, Planungs- und Entwicklungsarbeiten und Oeffentlichkeitsarbeit. Des weiteren werden die Veraenderungen sowie der Stand des CMB-Modellprojekts genannt. Im Anschluss werden die Ergebnisse der Evaluation im Berichtszeitraum praesentiert. Erwaehnung finden dabei (1) die Konzeption von standortuebergreifenden und Regionalveranstaltungen bzw. (2) Ergebnisse des ersten standortuebergreifenden CMB-Workshops zur Netzwerk- und Oeffentlichkeitsarbeit. Daran knuepfen die Ergebnisse der summativen Evaluation im Berichtszeitraum an. Sie gliedern sich in (1) eine Uebersicht ueber wichtige Merkmale der neu in das Modellprojekt aufgenommenen CMB-Stellen, (2) statistische Angaben zu Teilnehmerstruktur, Ablauf und Ergebnis des CMB sowie (3) die Probleme der CMB-Stellen im Berichtszeitraum. Der Text schliesst mit einem Ausblick, der die im Rahmen des Modellprojekts zu erledigenden Routineaufgaben (laufende Sichtung und Analyse themenbezogener Informationen aus Literatur und Internet usw.) umfasst. (ICG2)SIGLEAvailable from http://www.uni-erlangen.de/docs/FAU/anstalten/ifes/pub/pdf/m 5 2003.pdf / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekDEGerman

Ultraflexible Nanowire Array for Label- and Distortion-Free Cellular Force Tracking

Living cells interact with their immediate environment by exerting mechanical forces, which regulate important cell functions. Elucidation of such force patterns yields deep insights into the physics of life. Here we present a top-down nanostructured, ultraflexible nanowire array biosensor capable of probing cell-induced forces. Its universal building block, an inverted conical semiconductor nanowire, greatly enhances both the functionality and the sensitivity of the device. In contrast to existing cellular force sensing architectures, microscopy is performed on the nanowire heads while cells deflecting the nanowires are confined within the array. This separation between the optical path and the cells under investigation excludes optical distortions caused by cell-induced refraction, which can give rise to feigned displacements on the 100 nm scale. The undistorted nanowire displacements are converted into cellular forces via the nanowire spring constant. The resulting distortion-free cellular force transducer realizes a high-resolution and label-free biosenor based on optical microscopy. Its performance is demonstrated in a proof-of-principle experiment with living Dictyostelium discoideum cells migrating through the nanowire array. Cell-induced forces are probed with a resolution of 50 piconewton, while the most flexible nanowires promise to enter the 100 femtonewton realm.publishe

Pyridoxine responsiveness in novel mutations of the PNPO gene

OBJECTIVE: To determine whether patients with pyridoxine-responsive seizures but normal biomarkers for antiquitin deficiency and normal sequencing of the ALDH7A1 gene may have PNPO mutations. METHODS: We sequenced the PNPO gene in 31 patients who fulfilled the above-mentioned criteria. RESULTS: We were able to identify 11 patients carrying 3 novel mutations of the PNPO gene. In 6 families, a homozygous missense mutation p.Arg225His in exon 7 was identified, while 1 family was compound heterozygous for a novel missense mutation p.Arg141Cys in exon 5 and a deletion c.279_290del in exon 3. Pathogenicity of the respective mutations was proven by absence in 100 control alleles and expression studies in CHO-K1 cell lines. The response to pyridoxine was prompt in 4, delayed in 2, on EEG only in 2, and initially absent in another 2 patients. Two unrelated patients homozygous for the p.Arg225His mutation experienced status epilepticus when switched to pyridoxal 5'-phosphate (PLP). CONCLUSIONS: This study challenges the paradigm of exclusive PLP responsiveness in patients with pyridoxal 5'-phosphate oxidase deficiency and underlines the importance of consecutive testing of pyridoxine and PLP in neonates with antiepileptic drug-resistant seizures. Patients with pyridoxine response but normal biomarkers for antiquitin deficiency should undergo PNPO mutation analysis