An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolys...

Frequency Domain (FD) fluorimetry, capitalizes on the frequency response function of a fluorophore and offers independence from light scatter and excitation/emission intensity variations in order to extract the sample's fluorescent lifetime. Mercury vapor lamps, a common source of industrial facility lighting, emit radiation that overlaps the UV/blue absorption spectrum of many fluorophores and...

We report the application of a microfluidic device for semi-contact temperature measurement in picoliter volumes of aqueous media. Our device, a freely positionable multifunctional pipette, operates by a hydrodynamic confinement principle, i.e., by creating a virtual flow cell of micrometer dimensions within a greater aqueous volume. We utilized two fluorescent rhodamines, which exhibit differe...

Viscosity is a fundamental physical parameter that influences diffusion in biological processes. The distribution of intracellular viscosity is highly heterogeneous, and it is challenging to obtain a full map of cellular viscosity with detailed organelle information. In this work, we report 1 as the first fluorescent viscosity probe which is able to quantitatively map cellular viscosity with de...

The pH dependence of the binding of warfarin to human serum albumin has been studied by circular dichroism, fluorescence, and equilibrium dialysis. The pH dependences of the induced ellipticity and the fluorescent intensity of the warfarinalbumin complexes at low drug to protein ratios parallel the neutral to base transition, occurring in the protein over the pH range 6 to 9. Dialysis data show...

Fluorescent signal intensities from confocal laser scanning microscopes (CLSM) suffer from several distortions inherent to the method. Namely, these are the decay of intensities with increasing depth by finite transparency of the media used, the effects of extinction of the examined objects, the bleaching of the fluorochrome and stray light at surfaces within the volume under research. Under ce...

We present a general and straightforward one-step approach to enhance the photophysical properties of conjugated polyelectrolytes. Upon complexation with an amphiphilic polymer (polyvinylpyrrolidone), an anionic conjugated polyelectrolyte (poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylenevinylene]) was prepared into small nanoparticles with exceptional photostability and brightness. The polymer f...

One-dimensional photonic crystal PC slabs are capable of enhancing the excitation of fluorescent material adsorbed on their surface. In this report, we demonstrate and verify by electromagnetic computer simulations that resonant leaky modes spectrally overlapping the laser wavelength used for fluorescent excitation are responsible for the enhanced excitation, and that the Q-factor of the PC res...

A method of fluorescent nanoparticle-based indirect immunofluorescence microscopy (FNP-IIFM) was developed for the rapid detection of Mycobacterium tuberculosis. An anti-Mycobacterium tuberculosis antibody was used as primary antibody to recognize Mycobacterium tuberculosis, and then an antibody binding protein (Protein A) labeled with Tris(2,2-bipyridyl)dichlororuthenium(II) hexahydrate (RuBpy...

There is a pressing need in neuroscience for genetically-encoded, fluorescent voltage probes that can be targeted to specific neurons and circuits to allow study of neural activity using fluorescent imaging. We created 90 constructs in which the voltage sensing portion (S1-S4) of Ciona intestinalis voltage sensitive phosphatase (CiVSP) was fused to circularly permuted eGFP. This led to Electric...