RESUMO
In this paper, we studied the lateral deformation of human red blood cells (RBCs) during lateral indentation by an optically trapped silica bead with a diameter of 4.5 µm (Bangs Laboratories, Inc. Fishers, IN, USA). The images were captured using a CCD camera and the Boltzmann statistics method was used for force calibration. Using the Hertz model, we calculated and compared the elastic stiffness resulting from the lateral force, showing that the differences are important and that the force should be considered. Besides the lateral component, this setup also allowed us to examine the lateral cell-bead interaction. The mean values of the cell shear stiffness measured during indentation were 3.37 ± 0.40 µN/m for biconcave RBCs, 3.48 ± 0.23 µN/m for spherical RBCs, and 3.80 ± 0.22 µN/m for crenelated RBCs, respectively. These results show that this approach can be used as a routine method for RBC study, because it enabled us to manipulate the cell without contact with the wall.
RESUMO
We propose a new approach to calculate the sensitivity factor of the detector in optical tweezers. In this work, we used a charge-coupled device (CCD) camera and a quadrant photodiode (QPD) for the extraction of the various positions occupied by the trapped object (in this case, silica beads of different diameters). Image-J software and the Boltzmann statistical method were then used to estimate the sensitivity of the detector. Silica beads of diameter 0.8 µm, 2 µm, a system of 2 µm bead stuck to 4.5 µm one and another system of 2 µm beads stuck to 2 µm one, were studied. This work contributes significantly to making better calibration of the detector without taking into account the geometry of the object imprisoned in the optical trap. We further developed an approach to calculate the interaction force between two microbeads. This approach does not require any knowledge of solvent viscosity and works for all types of samples.