Reducing data acquisition for light‐sheet microscopy by extrapolation between imaged planes

Light‐sheet fluorescence microscopy (LSFM) is a powerful technique that can provide high‐resolution images of biological samples. Therefore, this technique offers significant improvement for three‐dimensional (3D) imaging of living cells. However, producing high‐resolution 3D images of a single cell or biological tissues, normally requires high acquisition rate of focal planes, which means a large amount of sample sections. Consequently, it consumes a vast amount of processing time and memory, especially when studying real‐time processes inside living cells. We describe an approach to minimize data acquisition by interpolation between planes using a phase retrieval algorithm. We demonstrate this approach on LSFM data sets and show reconstruction of intermediate sections of the sparse samples. Since this method diminishes the required amount of acquisition focal planes, it also reduces acquisition time of samples as well. Our suggested method has proven to reconstruct unacquired intermediate planes from diluted data sets up to 10× fold. The reconstructed planes were found correlated to the original preacquired samples (control group) with correlation coefficient of up to 90%. Given the findings, this procedure appears to be a powerful method for inquiring and analyzing biological samples. Gerchberg‐Saxton algorithm for phase retrieval is modified and applied for three‐dimensional (3D) objects reconstruction by extrapolation between Z‐axis diluted captures of two‐dimensional images. The modified algorithm is applied on 3D z‐stack biological samples data sets, acquired by light‐sheet microscopy. Implementation of the model is made on diluted data set as a proof of concept. By using up to ×10 dilution in data set, experimental results show reconstruction of interplane images with high correlation to the original images. This innovative method is therefore capable of reducing the required data acquisition rate and diminishes potential damage to samples.