A time-gated fluorescence detector using a tuning fork chopper.

We describe an affordable gated fluorescence detection system to measure fluorescent compounds with long-lifetimes that uses a tuning fork chopper to block the intense excitation pulse from a flash lamp or short-lived fluorescence. A conventional non-gated inexpensive photosensor module is used to collect the luminescence signal. Using the long-lived luminescence from the terbium(III)-dipicolinic acid (DPA) chelate, we demonstrate a limit of detection (LOD) of 120pM for DPA. This system is not only an order of magnitude less expensive than an electronically gateable phototomultiplier tube (GPMT), it exhibits no evidence of gradual loss of sensitivity, due likely to photocathode fatigue and deterioration, observed with a GPMT.

Nanofluidic channels by anodic bonding of amorphous silicon to glass to study ion-accumulation and ion-depletion effect.

A unique phenomenon, ion-enrichment and ion-depletion effect, exists in nanofluidic channels and is observed in amorphous silicon (alpha-Si) nanochannels as shallow as 50 nm. As a voltage is applied across a nanochannel, ions are rapidly enriched at one end and depleted at the other end of the nanochannel. alpha-Si is deposited on glass by plasma enhanced chemical vapor deposition and is selectively etched to form nanochannels. The depth of nanochannels is defined by the thickness of the alpha-Si layer. Low temperature anodic bonding of alpha-Si to glass was used to seal the channel with a second glass wafer. The strength of the anodic bond was optimized by the introduction of a silicon nitride adhesion promoting layer and double-sided bonding resulting from the electric field reversal. Completed channels, 50 nm in depth, 5 micron wide, and 1 mm long were completely and reliably sealed. Structures based on nanochannels 50-300 nm deep were successfully incorporated into nanofluidic devices to investigate ionic accumulation and depletion effect due to overlapping of electric double layer.

Mid-ultraviolet light-emitting diode detects dipicolinic acid.

Dipicolinic acid (DPA, 2,6-pyridinedicarboxylic acid) is a substance uniquely present in bacterial spores such as that from anthrax (B. anthracis). It is known that DPA can be detected by the long-lived fluorescence of its terbium chelate; the best limit of detection (LOD) reported thus far using a large benchtop gated fluorescence instrument using a pulsed Xe lamp is 2 nM. We use a novel AlGaN light-emitting diode (LED) fabricated on a sapphire substrate that has peak emission at 291 nm. Although the overlap of the emission band of this LED with the absorption band of Tb-DPA (lambda(max) doublet: 273, 279 nm) is not ideal, we demonstrate that a compact detector based on this LED and an off-the-shelf gated photodetection module can provide an LOD of 0.4 nM, thus providing a basis for convenient early warning detectors.

A nanoinjector for microanalysis.

We describe a simple miniature injection device that can be used for introduction of nanoliter sample volumes in microfluidic systems. The hybrid microstructure consists of two hydraulically connected parts, a pulse micropump, and a multilevel cross-flow injector. Sample injection is accomplished by creating a transient pressure pulse in the sample line by means of the solenoid-based micropump. The sample line is aligned at right angles to the main carrier flow line. The two flow channels are located in two different parallel planes. The cross section of the two channels is defined by a self-sealing aperture in an elastomer. During the pressure pulse, the sample is introduced through this aperture directly into the main flow stream. Fast impulse-based injection causes rapid mixing of the injected sample with the main flow stream. This permits simple single-line manifold micro flow injection (MFI) systems. The deformation/relaxation of the elastomer is fast and repeatable; as such, rapid serial actuations essentially result in a larger injected sample volume without significantly affecting the peak shape. In the present form, 2-40-nL samples are easily injected by single injection, and the injected volume can be chosen by system parameters. The injection repeatability as observed by a photometric detector is better than 1.2% (n = 100).

A microfabricated amperometric moisture sensor.

We describe a microfabricated moisture sensor with interdigitated Au or Pt electrodes on a silicon substrate. The sensor active area is covered with a spin-coated, baked-on layer of Nafion(R) perfluorosulfonate ionomer of submicron thickness. The sensor responds to moisture with a 10-90% rise time of 50-100 ms and a 90-10% fall time of 20-30 ms, faster than any other presently available sensor. The logarithm of the sensor current is related to the cube root of the moisture level at a given temperature. At 23 degrees C, the sensor easily measures relative humidities as low as 10%. The sensor response at a given absolute humidity level decreases exponentially with increasing temperature. The film is stable up to a temperature of 150 degrees C, permitting elevated temperature moisture measurement. Since sorbed water is actively decomposed electrolytically, the sensors exhibit negligible hysteresis. Response reproducibility of an individual sensor is <1%, that between identically made sensors is <5%, suggesting mass production techniques without individual calibration will be acceptable for all but the most demanding situation.