RESUMO
We present a real-time method to measure the amplitude of thermal fluctuations in biological membranes by means of a new treatment of the defocusing microscopy (DM) optical technique. This approach was also applied to study the deformation of human erythrocytes to its echinocyte structure. This was carried out by making three-dimensional shape reconstructions of the cell and measuring the thermal fluctuations of its membrane, as the cell is exposed to the anti-inflammatory drug naproxen and as it recovers its original shape, when it is subsequently cleansed of the drug. The results showed biomechanical changes in the membrane even at low naproxen concentration (0.2 mM). Also, we found that when the cell recovered its original shape, the membrane properties were different compared to the nondrugged initial erythrocyte, indicating that the drug administration-recovery process is not completely reversible.
Assuntos
Membrana Eritrocítica/fisiologia , Eritrócitos/citologia , Microscopia/métodos , Anti-Inflamatórios não Esteroides/farmacologia , Forma Celular , Membrana Eritrocítica/efeitos dos fármacos , Membrana Eritrocítica/patologia , Eritrócitos/patologia , Eritrócitos/fisiologia , Humanos , Processamento de Imagem Assistida por Computador , Naproxeno/farmacologia , TemperaturaRESUMO
High-speed cameras are reliable alternatives for the direct characterization of optical trap force and particle motion in optical tweezers setups, replacing indirect motion measurements often performed by quadrant detectors. In the present approach, subpixel motion data of the trapped particle is retrieved from a high-speed low-resolution video sequence. Due to the richness structure of motion diversity of microscopic trapped particles, which are subjected to a Brownian motion, we propose to also use the obtained motion information for tackling the inherent lack of resolution by applying superresolution algorithms on the low-resolution image sequence. The obtained results both for trapping calibration beads and for living bacteria show that the proposed approach allows the proper characterization of the optical tweezers by obtaining the real particle motion directly from the image domain, while still providing high resolution imaging.