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Biomedical subjects

Laurie E Locascio

Publications and source records attributed to Laurie E Locascio.

5 recordsLinked to original sources

Effect of caged fluorescent dye on the electroosmotic mobility in microchannels.

We report on measurements of electroosmotic mobility in polymer microchannels and silica capillaries with and without the addition of a caged fluorescein dye to the buffer. For PMMA microchannels, the mobility was found to increase from (2.6 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) to (4.6 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) upon addition of 1.2 mmol/L of caged dye. For PC microchannels, the mobility increased from (4.3 +/- 0.2) x 10(-4) cm2 V(-1) s(-1) to (5.4 +/- 0.1) x 10(-4) cm2 V(-1) s(-1) upon addition of caged dye. For PDMS microchannels, the mobility increased from (4.3 +/- 0.2) x 10(-4) cm2 V(-1) s(-1) to (6.4 +/- 0.5) x 10(-4) cm2 V(-1) s(-1) upon addition of caged dye. For fused-silica capillaries, the mobility ((5.5 +/- 0.2) x 10(-4) cm2 V(-1) s(-1)) was unaffected by the addition of the caged dye.

Journal Article↗

Microfluidic temperature gradient focusing.

A new technique is described for the concentration and separation of ionic species in solution within microchannels or capillaries. Concentration is achieved by balancing the electrophoretic velocity of an analyte against the bulk flow of solution in the presence of a temperature gradient. With an appropriate buffer, the temperature gradient can generate a corresponding gradient in the electrophoretic velocity, so that the electrophoretic and bulk velocities sum to zero at a unique point, and the analyte will be focused at that point. The technique is demonstrated for a variety of analytes, including fluorescent dyes, amino acids, DNA, proteins, and particles, and is shown to be capable of greater than 10,000-fold concentration of a dilute analyte.

Journal Article↗

Rapid microfluidic mixing.

A preformed T-microchannel imprinted in polycarbonate was postmodified with a pulsed UV excimer laser (KrF, 248 nm) to create a series of slanted wells at the junction. The presence of the wells leads to a high degree of lateral transport within the channel and rapid mixing of two confluent streams undergoing electroosmotic flow. Several mixer designs were fabricated and investigated. All designs were relatively successful at low flow rates (0.06 cm/s, > or = 75% mixing), but had varying degrees of success at high flow rates (0.81 cm/s, 45-80% mixing). For example, one design operating at high flow rates was able to split an incoming fluorescent stream into two streams of varying concentrations depending on the number of slanted wells present. The final mixer design was able to overcome stream splitting at high flow rates, and it was shown that the two incoming streams were 80% mixed within 443 microm of the T-junction for a flow rate of 0.81 cm/s. Without the presence of the mixer and at the same high flow rate, a channel length of 2.3 cm would be required to achieve the same extent of mixing when relying upon molecular diffusion entirely, while 6.9 cm would be required for 99% mixing.

Equipment Design↗

Control of electroosmotic flow in laser-ablated and chemically modified hot imprinted poly(ethylene terephthalate glycol) microchannels.

The fabrication of microchannels in poly(ethylene terephthalate glycol) (PETG) by laser ablation and the hot imprinting method is described. In addition, hot imprinted microchannels were hydrolyzed to yield additional charged organic functional groups on the imprinted surface. The charged groups are carboxylate moieties that were also used as a means for the further reaction of different chemical species on the surface of the PETG microchannels. The microchannels were characterized by fluorescence mapping and electroosmotic flow (EOF) measurements. Experimental results demonstrated that different fabrication and channel treatment protocols resulted in different EOF rates. Laser-ablated channels had similar EOF rates (5.3+/-0.3 x 10(-4) cm(2)/Vs and 5.6+/-0.4 x 10(-4) cm(2)/Vs) to hydrolyzed imprinted channels (5.1+/-0.4 x 10(-4) cm(2)/Vs), which in turn demonstrated a somewhat higher flow rate than imprinted PETG channels that were not hydrolyzed (3.5+/-0.3 x 10(-4) cm(2)/Vs). Laser-ablated channels that had been chemically modified to yield amines displayed an EOF rate of 3.38+/- 0.1 x 10(-4) cm(2)/Vs and hydrolyzed imprinted channels that had been chemically derivatized to yield amines showed an EOF rate of 2.67+/-0.6 cm(2)/Vs. These data demonstrate that surface-bound carboxylate species can be used as a template for further chemical reactions in addition to changing the EOF mobility within microchannels.

Electrophoresis, Capillary↗

Liposomes as signal amplification reagents for bioassays in microfluidic channels.

Liposomes with encapsulated carboxyfluorescein were used in an affinity-based assay to provide signal amplification for small-volume fluorescence measurements. Microfluidic channels were fabricated by imprinting in a plastic substrate material, poly(ethylene terephthalate glycol) (PETG), using a silicon template imprinting tool. Streptavidin was linked to the surface through biotinylated-protein for effective immobilization with minimal nonspecific adsorption of the liposome reagent. Lipids derivatized with biotin were incorporated into the liposome membrane to make the liposomes reactive for affinity assays. Specific binding of the liposomes to microchannel walls, dependence of binding on incubation time, and nonspecific adsorption of the liposome reagent were evaluated. The results of a competitive assay employing liposomes in the microchannels are presented.

Electrophoresis, Capillary↗