Size-dependent steady state saturation limit in biomolecular transport through nuclear membranes.

The nucleus preserves the genomic DNA of eukaryotic organisms and maintains the integrity of the cell by regulating the transport of molecules across the nuclear membrane. It is hitherto assumed that small molecules having a size below the passive permeability limit are allowed to diffuse freely to the nucleus while the transport of larger molecules is regulated via an active mechanism involving energy. Here we report on the kinetics of nuclear import and export of dextran molecules having a size below the passive permeability limit. The studies carried out using time-lapse confocal fluorescence microscopy show a clear deviation from the passive diffusion model. In particular, it is observed that the steady-state concentration of dextran molecules inside the nucleus is consistently less than the concentration outside, in contradiction to the predictions of the passive diffusion model. Detailed analysis and modeling of the transport show that the nuclear export rates significantly differ from the import rates, and the difference in rates is dependent on the size of the molecules. The nuclear export rates are further confirmed by an independent experimental study where we observe the diffusion of dextran molecules from the nucleus directly. Our experiments and transport model would suggest that the nucleus actively rejects exogenous macromolecules even below the passive permeability limit. This result can have a significant impact on biomedical research, especially in areas related to targeted drug delivery and gene therapy.

Confocal imaging of single BaTiO3 nanoparticles by two-photon photothermal microscopy.

We report on the development of a nonlinear optical microscopic technique based on two-photon absorption induced photothermal effect capable of detecting individual nonfluorescent nanoparticles with high sensitivity. The method which is inherently confocal makes use of near infrared excitation at high repetition rates and would be of interest in deep tissue imaging. We demonstrate the applicability of the technique by imaging single BaTiO3 nanoparticles, a potential biomolecular label having high photostability, in a scattering environment at fast time scales with a pixel dwell time of 80 µs.

Laser scanning photothermal microscopy: fast detection and imaging of gold nanoparticles.

We report on the design and construction of a laser scanning photothermal microscope and present images of gold nanoparticles of size as small as 5 nm. Laser scanning method allows fast image acquisition at 80 µs pixel dwell time so that a 500 × 500 pixel image is acquired in 20 s. Photothermal imaging at fast time scales can have potential applications in variety of fields including tracking of biomolecular transport processes.