Bioactivity Profiling of Small-Volume Samples by Nano Liquid Chromatography Coupled to Microarray Bioassaying Using High-Resolution Fractionation.

High-throughput screening platforms for the identification of bioactive compounds in mixtures have become important tools in the drug discovery process. Miniaturization of such screening systems may overcome problems associated with small sample volumes and enhance throughput and sensitivity. Here we present a new screening platform, coined picofractionation analytics, which encompasses microarray bioassays and mass spectrometry (MS) of components from minute amounts of samples after their nano liquid chromatographic (nanoLC) separation. Herein, nanoLC was coupled to a low-volume liquid dispenser equipped with pressure-fed solenoid valves, enabling 50-nL volumes of column effluent (300 nL/min) to be discretely deposited on a glass slide. The resulting fractions were dried and subsequently bioassayed by sequential printing of nL-volumes of reagents on top of the spots. Unwanted evaporation of bioassay liquids was circumvented by employing mineral oil droplets. A fluorescence microscope was used for assay readout in kinetic mode. Bioassay data were correlated to MS data obtained using the same nanoLC conditions in order to assign bioactives. The platform provides the possibility of freely choosing a wide diversity of bioassay formats, including those requiring long incubation times. The new method was compared to a standard bioassay approach, and its applicability was demonstrated by screening plasmin inhibitors and fibrinolytic bioactives from mixtures of standards and snake venoms, revealing active peptides and coagulopathic proteases.

A lightweight telemetry system for recording neuronal activity in freely behaving small animals.

A miniature lightweight radio telemetric device is described which is shown to be suitable for recording neuronal activity in freely behaving animals. Its size (12 x 5 x 8 mm) and weight (1.0-1.1 g with batteries, 0.4-0.5 g without) make the device particularly suitable for recording neuronal units in small animals such as mice or zebra finches. The device combines a high impedance preamplifier, RC-filters and an FM-transmitter. Using the device we recorded action potentials in field L of freely behaving zebra finches (12-17 g) through chronically implanted tungsten electrodes. In freely behaving birds we observed frequency dependent responses of field L units to auditory stimuli for periods of up to 7 days. We investigated the effect of the device on singing and locomotor activity of the zebra finches. Singing and locomotion were significantly affected on the first day after surgery. Both anesthesia and the presence of the transmitter contributed to the observed effect. After 1 day of recovery, singing activity returned to 99.6% and perch-hopping activity to 55.3% of the baseline levels. It is concluded that the device is well suited for recording spike trains from small animals while they behave freely and naturalistically.