Analyzing subcellular structure with optical Fourier filtering based on Gabor filters

Label-free measurement of subcellular morphology can be used to track dynamically cellular function under various conditions and has important applications in cellular monitoring and in vitro cell assays. We show that optical filtering of scattered light by two-dimensional Gabor filters allows for direct and highly sensitive measurement of sample structure. The Gabor filters, which are defined by their spatial frequency, orientation and Gaussian envelope, can be used to track locally and in situ the characteristic size and orientation of structures within the sample. Our method consists of sequentially implementing a set of Gabor filters via a spatial light modulator placed in a conjugate Fourier plane during optical imaging and identifying the filters that yield maximum signal. Using this setup, we show that Gabor filtering of light forward-scattered by spheres yields an optical response which varies linearly with diameter between 100nm and 2000nm. The optical filtering sensitivity to changes in diameter is on the order of 20nm and can be achieved at low image resolution. We use numerical simulations to demonstrate that this linear response can be predicted from scatter theory and does not vary significantly with changes in refractive index ratio. By applying this Fourier filtering method in samples consisting of diatoms and cells, we generate false-color images of the object that encode at each pixel the size of the local structures within the object. The resolution of these encoded size maps in on the order of 0.36μm. The pixel histograms of these encoded images directly provide 20nm resolved "size spectra", depicting the size distribution of structures within the analyzed object. We use these size spectra to differentiate the morphology of apoptosis-competent and bax/bak null apoptosis-resistant cells during cell death. We also utilize the sensitivity of the Gabor filters to object orientation to track changes in organelle morphology, and detect mitochondrial fission in cells undergoing apoptosis.