Super-High-Resolution Densitometry by Optical Trapping of a Single-Molecularly Imprinted Polymer Particle for Subsingle Molecule Detection.

In this study, for the first time, we have shown that the single nano-/microparticle trapping ability of optical tweezers combined with the high selectivity of molecularly imprinted polymers (MIPs) provides an indispensable molecular-level instrument for chemical sciences. Trapping a single MIP inside a solution and analyzing its Brownian motion allow for real-time determination of its target molecule [trimipramine (TMP) in our case] content. This method is also utilized to precisely measure TMP concentration in the bulk solution. The detection and optical volumes, respectively, defined as single MIP volume and laser focal volume, were about a few femtoliters. According to our data within a detection volume inside the bulk solution, 0.02-0.25 target molecules could be detectable with a detection limit of 0.005 molecules. Thus, we detected 1/1000th of the subsingle molecule in detection volume by high-resolution densitometry.

Fluorescence resonance energy transfer-thermal lens spectrometry (FRET-TLS) as molecular counting of methamphetamine.

A novel and sensitive approach has been presented for the determination of methamphetamine (METH) based on fluorescence resonance energy transfer-thermal lens spectrometry (FRET-TLS). Due to the affinity of fluorescein molecules to the surface of AuNPs through the electrostatic interaction and thereby caused reduction of the distance between fluorescein and AuNPs, the best way for de-excitation of excited fluorescein is FRET. The energy absorbed by fluorescein transferred to AuNPs causes enhancement of the thermal lens effect. The thermal lens of the fluorescence molecule could be enhanced through a proper acceptor. Upon the addition of methamphetamine, the fluorescein molecules are detached from the surface of AuNPs, due to the stronger adsorption of methamphetamine. As a result, the fluorescence of fluorescein recovered, and the thermal lens effect of fluorescein decreased. The mechanism of energy transfer was evaluated by two different methods including time-resolved spectroscopy and thermal lens spectrometry. Under the optimal conditions, the thermal lens signal was linearly proportional to methamphetamine concentration in the range 5 - 80 nM. The limit of detection and limit of quantitation were 1.5 nM and 4.5 nM, respectively. The detection volume and limit of molecules in the detection volume were 960 attoliter and 87 molecules, respectively. The method was successfully applied for the determination of methamphetamine in human blood plasma and urine.

Enhancmentation of Photo-Thermal Lens of Fluorescence Molecules by Fluorescence Resonance Energy Transfer Mechanism.

The photo-thermal lens technique is based on the measurement of the gradient temperature that is produced in an illuminated sample by laser beam as a result of non-radiative relaxation. The sensitivity of photo-thermal lens is higher than conventional absorption techniques, due to the direct measurement of absorbed energy. However, the main drawback of the photo-thermal lens technique is the inability to measure molecules that release their excess energy of absorption in the form of radiation relaxation. In fluorescence molecules, because their excess energy is released by radiation, the photo-thermal lens is insignificant. In this research, we aim to increase the photo-thermal lens sensitivity of the fluorescence molecules by selecting the appropriate acceptor for the fluorescence molecules as the donor. We investigated the effect of the presence of AuNPs on the photo-thermal lens of di-triazene (DTA) as fluorescence molecules. We observed that AuNPs enhanced the photo-thermal lens of DTA. The quantum yield of DTA was calculated to be 90.45%. The energy transfer from DTA to AuNPs occurs by fluorescence resonance energy transfer (FRET) mechanism. The FRET efficiency was 83.43%. In the presence of AuNPs, the photo-thermal lens of the DTA was amplified by four times more. The calibration curve of DTA is linear in the range of 0.1 to 110 ng/mL. The limited of detection (LOD) value of 0.08 ng/mL is calculated.