Use of optophoresis as an in vitro predictor of cell response to chemotherapy for chronic lymphocytic leukemia.

B-cell chronic lymphocytic leukemia [CLL] is characterized by active accumulation of clonal CD5+/CD19+/CD23+ B cells. Individualized characterization of patient cell resistance/sensitivity to specific agents can provide important information to guide therapy selection. We have utilized optophoresis, which is a technique for the analysis of the motion of cells within a moving optical gradient field. It detects the broad cellular changes associated with apoptosis based on physical characteristics of the cell, such as morphology, size, refractive index, density, and surface properties. We analyzed peripheral blood samples from 62 CLL patients in the presence of varying concentrations of chemotherapeutic agents. Optophoresis and a more conventional measurement of cell death were utilized. The outcome of ex vivo drug resistance using optophoresis was compared to clinical response in 30 patients for which there was clinical outcome data available. The overall accuracy of optophoresis in reflecting clinical response was 80%. It has advantages over alternative methods of determining chemoresistance including the ability to evaluate very small sample sizes and ability to work in mixed-cell populations. Changes in cell physical characteristics in response to chemotherapy, as measured by optophoresis is an accurate method for predicting chemosensitivity ex vivo in CLL.

Elastic light scattering from single cells: orientational dynamics in optical trap.

Light-scattering diagrams (phase functions) from single living cells and beads suspended in an optical trap were recorded with 30-ms time resolution. The intensity of the scattered light was recorded over an angular range of 0.5-179.5 degrees using an optical setup based on an elliptical mirror and rotating aperture. Experiments revealed that light-scattering diagrams from biological cells exhibit significant and complex time dependence. We have attributed this dependence to the cell's orientational dynamics within the trap. We have also used experimentally measured phase function information to calculate the time dependence of the optical radiation pressure force on the trapped particle and show how it changes depending on the orientation of the particle. Relevance of these experiments to potential improvement in the sensitivity of label-free flow cytometry is discussed.