We have developed a multiplexed holographic optical tweezers system with an imaging spectrometer to manipulate multiple optically trapped nanosensors and detect multiple fluorescence spectra. The system uses a spatial light modulator (SLM) to control the positions of infrared optical traps in the sample so that multiple nanosensors can be positioned into regions of interest. Spectra of multiple nanosensors are detected simultaneously with the application of an imaging spectrometer. Nanosensors are capable of detecting changes in their environment such as pH, ion concentration, temperature, and voltage by monitoring changes in the nanosensors' emitted fluorescence spectra. We use streptavidin labeled quantum dots bound to the surface of biotin labeled polystyrene microspheres to measure temperature changes by observing a corresponding shift in the wavelength of the spectral peak. The fluorescence is excited at 532 nm with a wide field source.
Spatial light modulators (SLMs) are used to diffract light beams for a variety of applications. In particular, optical tweezer trapping has greatly benefited from the advent of phase-mostly and phase-only SLMs to write holograms that produce multiple traps. We are using holographic optical tweezers to trap multiple sensor particles in a lab-on-a-chip measurement platform. As part of this program, we have developed a method for optimizing the diffraction efficiency of a SLM. This general method can be applied in situ and addresses the issues of nonlinear phase modulation and phase modulation less than 2π. The method employs a one-dimensional blazed phase grating written on the SLM. For an ideal SLM, the phase shift is linear and covers 0 - 2π, yielding a first-order diffraction efficiency of unity. For a realistic SLM with nonlinear or reduced phase shift, the efficiency is approximately η = 1 - σ<sup>2</sup>, where σ<sup>2</sup> is the variance of the phase error from the ideal case. Because each pixel contributes to the phase error independently, this suggests a method to maximize the efficiency by adjusting the phase encoding of the SLM pixel-by-pixel. In practice, we do this by adjusting the gray-scale of each pixel while measuring the first-order diffracted power. The collection of optimal gray values comprises the optimized gray-scale lookup table, which exhibits the nonlinearity required to produce a linear phase grating and the saturated phase encoding that maximizes the efficiency of phase limited SLMs. We have successfully applied this optimization method to two different SLMs.