Victor Krivenkov

Two-photon-induced Förster resonance energy transfer in a hybrid material engineered from quantum dots and bacteriorhodopsin

Energy transfer from nanostructures to biological supramolecular photosystems is an important fundamental issue related to the possible influence of nanoobjects on biological functions. We demonstrate here two-photon-induced Förster resonance energy transfer (FRET) from fluorescent CdSe/ZnS quantum dots (QDs) to the photosensitive protein bacteriorhodopsin (bR) in a QD-bR hybrid material. The two-photon absorption cross section of QDs has been found to be about two orders of magnitude larger than that of bR. Therefore, highly selective two-photon excitation of QDs in QD-bR complexes is possible. Moreover, the efficiency of FRET from QDs to bR is sufficient to initiate bR photoconversion through two-photon excitation of QDs in the infrared spectral region. The data demonstrate that the effective spectral range in which the bR biological function is excited can be extended beyond the band where the protein itself utilizes light energy, which could open new ways to use this promising biotechnological material.

Surface ligands affect photoinduced modulation of the quantum dots optical performance

Changes of optical properties of the solutions of CdSe/ZnS quantum dots (QDs) covered with the trioctylphosphine oxide (TOPO) ligands under the pulsed ultraviolet (UV) laser irradiation are observed. The fluorescence quantum yield (QY) of QDs decreases by more than an order of magnitude when the radiation dose approaches 2 × 10-15 J per particle. This process is accompanied by a blue shift of both fluorescence and the first excitonic absorption peaks. The fluorescence quenching becomes less pronounced when the overall TOPO content in the solution is increased. When ТОРО ligands are replaced with n-hexadecylamine (HDA), QY and spectral properties are not changed at the same irradiation conditions. We assume that the above changes of the optical properties are associated with photooxidation of TOPO ligands by excited QD. Such process is less probable for the HDA ligand due to its different energy structure.

Resonance energy transfer in nano-bio hybrid structures can be modulated by UV laser irradiation

A method for targeted variation of the radiation properties of quantum dots (QDs) to control the efficiency of resonance energy transfer in nanocrystal assemblies and nano-bio hybrid materials has been developed. The method is based on strong ultraviolet (UV) laser irradiation of QDs and allows the extinction and luminescence spectra to be controlled and the luminescence quantum yield and decay kinetics to be varied. Water-soluble QDs have been synthesized and used for analyzing the effect of energy transfer from semiconductor nanocrystals on the photocycle of the photosensitive protein bacteriorhodopsin (bR) in bR–QD complexes. The UV irradiation mode has been selected in a way permitting the modulation of QD optical parameters without modification of their structure or physico-chemical properties. It is concluded that the QD interaction with bR accelerates its photocycle, but this acceleration is determined by electrostatic interactions, rather than Forster resonance energy transfer from QDs to bR. The method of UV laser irradiation of fluorescent semiconductor QDs has proven to be an efficient technique for variation of nanocrystal optical properties without affecting their structure, as well as for fine modulation of the energy transfer processes in the nanocrystal assemblies and nano-bio hybrid materials.

The influence of the quantum dot/polymethylmethacrylate composite preparation method on the stability of its optical properties under laser radiation

Photoluminescent semiconductor nanocrystals, quantum dots (QDs), are nowadays one of the most promising materials for developing a new generation of fluorescent labels, new types of light-emitting devices and displays, flexible electronic components, and solar panels. In many areas the use of QDs is associated with an intense optical excitation, which, in the case of a prolonged exposure, often leads to changes in their optical characteristics. In the present work we examined how the method of preparation of quantum dot/polymethylmethacrylate (QD/PMMA) composite influenced the stability of the optical properties of QD inside the polymer matrix under irradiation by different laser harmonics in the UV (355 nm) and visible (532 nm) spectral regions. The composites were synthesized by spin-coating and radical polymerization methods. Experiments with the samples obtained by spin-coating showed that the properties of the QD/PMMA films remain almost constant at values of the radiation dose below ~10 fJ per particle. Irradiating the composites prepared by the radical polymerization method, we observed a monotonic increase in the luminescence quantum yield (QY) accompanied by an increase in the luminescence decay time regardless of the wavelength of the incident radiation. We assume that the observed difference in the optical properties of the samples under exposure to laser radiation is associated with the processes occurring during radical polymerization, in particular, with charge transfer from the radical particles inside QDs. The results of this study are important for understanding photophysical properties of composites on the basis of QDs, as well as for selection of the type of polymer and the composite synthesis method with quantum dots that would allow one to avoid the degradation of their luminescence.

Engineering of Optically Encoded Microbeads with FRET-Free Spatially Separated Quantum-Dot Layers for Multiplexed Assays

Quantum dot (QD) encoded microbeads are emerging for multiplexed analysis of biological markers. The quantitative encoding of microbeads prepared with different concentrations of QDs of different colors suffers from resonance energy transfer from the QDs fluorescing at shorter wavelengths to the QDs fluorescing at longer wavelengths. Here, we used the layer-by-layer deposition technique to spatially separate QDs of different colors with several polymer layers so that the distance between them would be larger than the Förster energy transfer radius. We performed fluorescence lifetime measurements to investigate and determine the conditions excluding significant resonance energy transfer between QDs within QD-encoded microbeads. Additionally, the number of QDs adsorbed onto microbeads was systematically established and multilayer structures of the QD-encoded microbead shells were characterized by scanning probe nanotomography. Finally, we prepared eight populations of FRET-free microbeads encoded with QDs of three colors at two intensity levels and demonstrated that all the optical codes are excitable at a single wavelength and may be clearly identified in three channels of a flow cytometer. The developed approach for engineering QD-encoded microbeads that are free from optical artefacts related to inter-QD resonance energy transfer paves the way to quantitative QD-based multiplexed assays.

Controllable photo-brightening/photo-darkening of semiconductor quantum dots under laser irradiation

It has been demonstrated that photo-induced changes in the optical properties of semiconductor quantum dots (QDs) can be controlled by tuning the parameters of their laser irradiation to vary the relative contributions of photo-brightening and photo-darkening of QDs. For this purpose, the effects of the QD size, photon energy, and intensity of irradiation of QDs on the competing processes of photo-darkening and photo-brightening have been investigated. We have found that photo-brightening of QDs is not accompanied by detectable growth of their photoluminescence (PL) decay time, this process being most pronounced for QDs with an originally low PL quantum yield (QY). In this case, an increase in the PL QY is assumed to be caused by transition of some QDs from the dark (non-emissive) state to the bright (emissive) state. On the other hand, the photo-darkening effect, which was observed only under UV irradiation at 266 nm, was accompanied by simultaneous drop of both the QD QY and their PL decay time. We have also found that, at a constant dose of absorbed energy, the photo-brightening and photo-darkening processes do not depend on the excitation intensity. Thus, the photo-induced changes in the optical properties of QDs are one-photon processes. These data may help to optimize the QD operational conditions in practical applications requiring their intense excitation and add to understanding the fundamental mechanisms of the irreversible photo-induced changes that occur in colloidal QDs under illumination.

Photoinduced modification of quantum dot optical properties affects bacteriorhodopsin photocycle in a (quantum dot)- bacteriorhodopsin hybrid material

A method for controlled changes in the radiative properties of quantum dots (QDs) in order to modulate the Forster resonance energy transfer (FRET) rate in nano-hybrid materials is proposed. The mechanism underlying the effect of QDs with optical properties modulated by UV laser irradiation on the photocycle of the photosensitive protein bacteriorhodopsin (bR) in its native purple membranes (PM) isolated from Halobacterium salinarum has been studied. The irradiation leads to a twofold decrease in the QD fluorescence quantum yield without changes in the extinction spectrum or the position or shape of the fluorescence spectrum. The bR photocycle is accelerated, which has been shown to be related to the changes of the surface potential of PM upon formation of their complexes with QDs.

Laser tuning of resonance energy transfer efficiency in a quantum dot– bacteriorhodopsin nano–bio hybrid material

Semiconductor quantum dots (QDs) are a promising “nano-antennas” capable of absorbing efficiently light energy upon one- or two-photon excitation and then transferring it to convenient energy acceptors via Förster resonance energy transfer (FRET). The photosensitive protein bacteriorhodopsin (bR) has been shown to be a promising material for optoelectronic and photovoltaic applications, but it cannot effectively absorb light in the UV, blue, and NIR regions. It was shown previously that formation of hybrid complexes of QDs and purple membranes (PMs) containing bR could significantly improve the bR capacity for utilizing light upon one- and two-photon laser excitations. Under the laser irradiation, the optical properties of bR itself remain unchanged, whereas those of QDs may be altered. Therefore, it is important to study the effects of intense laser excitation on the properties of the QD–PM hybrid material. In this study we have shown that laser irradiation can lead to an increase in the luminescence quantum yield (QY) of QDs. The fact that this irradiation does not change the QD absorption spectra means that the QD quantum yield may be optically controlled without changing the QD structure or composition. Finally, we have shown experimentally that photoinduced increase in the QY of QDs lead to the corresponding increase in the efficiency of FRET in the QD–PM hybrid material. As a result, an approach to increasing the FRET efficiency in hybrid nano-biomaterials where QDs serve as donors have been proposed.

Resonant transfer of one- and two-photon excitations in quantum dot–bacteriorhodopsin complexes

Light-sensitive protein bacteriorhodopsin (BR), which is capable of electrical response upon exposure to light, is a promising material for photovoltaics and optoelectronics. However, the rather narrow absorption spectrum of BR does not allow achieving efficient conversion of the light energy in the blue and infrared spectral regions. This paper summarizes the results of studies showing the possibility of extending the spectral region of the BR function by means of the Förster resonance energy transfer (FRET) from CdSe/ZnS quantum dots (QDs), which have a broad spectrum of one-photon absorption and a large twophoton absorption cross section (TPACS), to BR upon one- and two-photon excitation. In particular, it is shown that, on the basis of QDs and BR-containing purple membranes, it is possible to create electrostatically associated bio-nano hybrid systems in which FRET is implemented. In addition, the large TPACS of QDs, which is two orders of magnitude larger than those of BR and organic dyes, opens up a means for selective two-photon excitation of synthesized bio-nano hybrid complexes. On the basis of the results of this work, the spectral region in which BR converts the light energy into electrical energy can be extended from the UV to near-IR region, creating new opportunities for the use of this material in photovoltaics and optoelectronics.

Use of semiconductor nanocrystals to encode microbeads for multiplexed analysis of biological samples

Microbeads encoded with semiconductor quantum dots (QDs) are suitable tools for multiplexed analyses of various biological markers using flow cytometry. We have prepared a panel of microbeads encoded with QDs of different colors emitting with different luminescence intensities using the layer-by-layer deposition technique, which consists in layering of alternately charged polyelectrolytes and negatively charged QDs onto the surface of microbeads. This method allows QDs to be separated with one or several polymer layers in order to prevent Forster resonance energy transfer (FRET) and the resultant quenching of QD fluorescence in multicolor microbeads.