Single-Shot High-Throughput Phase Imaging with Multibeam Array Interferometric Microscopy

Abstract

High-throughput microscopic imaging is highly desirable in biomedical applications. Advances in computational microscopy have achieved high space-bandwidth products and even permitted gigapixel imaging in a stepwise fashion, yet temporal resolution remains challenging for investigating live-sample dynamics. Here, we report multibeam array interferometric microscopy (MAIM) for a single-shot high space-bandwidth product. The MAIM method overcomes the limitations of conventional digital holographic microscopy, providing complex field reconstruction with a maximum 5-fold field of view (FOV) increase in a single camera acquisition, while maintaining sub-nanometer optical path-length stability. This is achieved by integrating common-path holographic microscopy, multibeam interference technology, and holographic multiplexing technology. The temporal resolving power of MAIM is significantly higher than that of computational illumination microscopy. MAIM has major advantages over previous holographic multiplexing techniques in that it integrates more wavefronts and offers high temporal stability. The fundamentals of MAIM are analyzed theoretically. As a demonstration, we build MAIM prototypes to increase the FOV by factors of 5, 4, and 3, respectively. We present proof-of-concept MAIM imaging results of both natural and artificial samples and show biomedical applications such as monitoring sub-cellular dynamical phenomena in flowing live erythrocytes in vitro and label-free microrefractometry imaging of unstained cancer tissue slices. MAIM gives rise to (ultra)­fast or long-term (time-lapse) imaging of nanoscale dynamics of unstained live samples in vitro with a high throughput.

Vector Differential Interference Contrast Microscopy Based on a 3-in-1 Phase Mask through a Dynamic Diffractive Optical Element

“…where c R * is the complex function calculated from the samplefree reference interference to remove the heterodyne carrier and the system phase residual. 8 Applying the algorithm across the entire camera plane, the x-differential phase map of the sample is finally retrieved. The y-differential phase map, as well as the 45°one, can be retrieved through a similar fashion by selecting corresponding heterodyne carrier.…”

“…Recalling the frequency spectrum shown in Figure f, a spectrum filter is used to isolate the heterodyne carrier at f 1 . Then the inverse Fourier transform is applied to the isolated spectrum C 1 ( f t – f 1 ) to obtain the complex function, denoted by The x -differential phase at this pixel is then calculated by where c R * is the complex function calculated from the sample-free reference interference to remove the heterodyne carrier and the system phase residual . Applying the algorithm across the entire camera plane, the x -differential phase map of the sample is finally retrieved.…”

“…Quantitative phase imaging (QPI) has been used extensively in the visualization of weakly scattering samples, especially for transparent biomedical specimens such as cells. , The retrieved phase information indicates the optical path length of the imaging samples, representing the samples’ morphological properties, and further links to its biological or chemical characteristics. − The label-free QPI involves neither staining nor fluorescence to avoid potential photobleaching and phototoxicity. , The rapid development of QPI involves digital holographic microscopy, tomographic QPI, Fourier ptychography, and quantitative differential interference contrast (DIC) microscopy. , …”

“…3−5 The label-free QPI involves neither staining nor fluorescence to avoid potential photobleaching and phototoxicity. 6,7 The rapid development of QPI involves digital holographic microscopy, 8 tomographic QPI, 9 Fourier ptychography, 10 and quantitative differential interference contrast (DIC) microscopy. 11,12 Over the past decade, the miniaturization of microscopes has opened up possibilities for in vivo bioimaging using portable devices.…”