|Alignment of 1H-NMR data using a Generalized Fuzzy Hough Transform|
Erik Alm, Leonard Csenki, Ralf J.O. Torgrip, K. Magnus Åberg, Lars I. Nord, Ina Schuppe-Koistinen and Johan Lindberg
In metabolic profiling, multivariate data analysis techniques are used to interpret 1D 1H–NMR data. Multivariate data analysis techniques require that peaks are located in the same variables in every spectrum – this requirement is not met in native NMR spectra. Current state-of-the-art alignment algorithms are unable to align peaks when the spatial order of the peaks changes. We present the Fuzzy Generalized Hough Transform alignment which solves the alignment problem.
|1H NMR for the Accurate Quantification of Analytical Reference Standards|
Jan Schripsema, Denise Dagnino and Peter Verhaert
NMR can be used for the quantification of minute quantities of standard compounds. The high specificity of NMR signals permits absolute quantifications independent of the presence of impurities. The feasibility and limits of NMR application are demonstrated through the analysis of algal toxins and polar substances in butter.
|Single Cell Analysis of Voltage-gated Potassium Channels for Electrophysiological Properties of Rat Hypothalamic Paraventricular Nucleus Neurons|
Seul Ki Lee, Sora Lee, Seung Yub Shin, Pan Dong Ryu, So Yeong Lee
The hypothalamic paraventricular nucleus (PVN), an integrating site in the regulation of neuroendocrine and autonomic nervous systems, is composed of heterogeneous neurons, type I and type II, based on their electrophysiological properties. Single cell real-time RT-PCR analysis was made to investigate the molecular basis underlying the properties. The results suggest that Kv1.3, Kv4.2 and Kv4.3 potassium channels are the potential candidates in determining the electrophysiological properties in
|Fragment Screening Using Capillary Electrophoresis|
Simon Pettit, Carol Austin, Dallas Hughes, Sharon Magnolo, Leonid Raskovetsky
Capillary electrophoresis, which was pioneered for HTS in the 1990s, has recently been adapted to enable the screening of fragments. The technique is explained and the screening of fragments against a chaperone protein Hsp90 is presented as an example.
|High-Throughput Multiplexed Inflammation Assay Using Primary Cells|
Oksana Sirenko, Yen-Wen Chen, Jayne Hesley, Cathy Olsen, H. Roger Tang, Evan F. Cromwell*
An important event in the inflammation response is the expression of cell-surface antigens that facilitate binding of immune cells to blood vessels. The ability to monitor up-regulation of these molecules, such as VCAM, E-selectin, and HLADR on endothelial cells provides an important physiological read-out for cell-based models of inflammation.
|Maximizing cell wash performance with the AquaMax® 2000/4000 microplate washers|
Cathy Olsen, Iris Yang, and James Wasson
Many cell-based applications, including calcium flux assays, cell-based ELISAs, ion channel assays, and others require gentle but thorough washing of cells in order to ensure cell retention, cell integrity and optimal assay performance. The 96 cell wash head for AquaMax® 2000/4000 microplate washers from Molecular Devices uses angled pins to dispense fluid without disturbing cells. Adjustable dispense rate and aspiration height allow users to minimize disruption of even weakly adherent cells. We
|Two secreted orthogonal bioluminescent reporters for non-destructive ultrasensitive assays|
George Tzertzinis1, Salvatore Russello1, Cathy Olsen2
Here we present a powerful combination of two secreted bioluminescent reporters, the luciferases from Gaussia princeps and Cypridina noctiluca. We demonstrate that the properties of these two reporters in terms of sensitivity, compatibility and use in non- destructive assays conducted on live cells or supernatants from different well formats makes them suitable for novel high throughput format applications.
|Label free technology: the weird and wonderful world of cell signaling|
Ryall A†, Shearer J†, Brodbeck D*, Nufer O*, Cronk D† and Fasler S*
Whilst conducting an evaluation of label free technology platforms we
encountered a number of surprising, but reproducible, observations. For
example, receptor over-expression impacts endogenous signaling
responses; surface coatings affecting cell adherence can change the
response profile; and it is important to choose the appropriate analysis
metric for complex profiles. Awareness of the subtleties of cell signaling
within model systems such as CHO and HEK recombinant over-expression
|Label free technology: value added information in the drug discovery process|
Brodbeck D*, Nufer O*, Ryall A†, Shearer J†, Fasler S* and Cronk D†
We have evaluated Molecular Devices CellKey™ and SRU BIND® label free
detection technologies and their application to hit finding and hit validation
using cell based assays. The presented data highlights: