Optical diffractive tomography (ODT) microscopy is a normal wide-field, non-invasive and label-free three-dimensional imaging technology for cells and tissues. The traditional ODT microscopy has a little field-of-view (FOV) of about 80um, which needs to be reconstructed by regions and then spliced for large complex biological samples with sub-millimeter scale. However, there is interference of ringing effect during splicing, which limits its application in sub-millimeter biological samples. In this paper, an ODT microscopy with a wider FOV is proposed. The FOV is 196um, more than three times that of conventional technology, and the photon flux is higher. The results show that the wider-field ODT microscopy has better imaging performance, higher signal-to-noise ratio on the sub-millimeter samples without splicing.
Chiral sum-frequency generation (SFG) has proven to be a versatile spectroscopic and imaging tool for probing chirality. However, due to polarization restriction, the conventional chiral SFG microscopes have mostly adopted noncollinear beam configurations, which only partially cover the aperture of microscope and strongly spoil the spatial resolution. In this study, we report the first experimental demonstration of collinear chiral SFG microscopy, which fundamentally supports diffraction-limited resolution. This advancement is attributed to the collinear focus of a radially polarized vectorial beam and a linearly polarized (LP) beam. The tightly focused vectorial beam has a very strong longitudinal component, which interacts with the LP beam and produces the chiral SFG. The collinear configuration can utilize the full aperture and thus push the spatial resolution close to the diffraction limit. This technique can potentially boost the understanding of chiral systems.
The emergence of super-resolution (SR) fluorescence microscopy has rejuvenated the search for new cellular sub-structures. However, SR fluorescence microscopy achieves high contrast at the cost of the lack of a holistic view of their interacting partners and surrounding environment. Thus we develop SR fluorescence-assisted diffraction computational tomography (SR-FACT), which combines label-free three-dimensional optical diffraction tomography (ODT) with two-dimensional fluorescence Hessian structured illumination microscopy. The ODT module is capable of resolving mitochondria, lipid droplets, the nuclear membrane, chromosomes, the tubular endoplasmic reticulum and lysosomes. Using dual-mode correlated live cell imaging for prolonged period of time, we observe the dynamics of a novel subcellular structure named dark-vacuole bodies. These works demonstrate the unique capabilities of SR-FACT, which suggest its wide applicability in cell biology in general.
A commentary on the article “Three-dimensional tomography of red blood cells using deep learning” by J. Lim, A. Ayoub, and D. Psaltis, Adv. Photonics Volume 2, Issue 2, doi: 10.1117/1.AP.2.2.026001.
There have been increasing interests in nonlinear optical imaging technology, especially in biomedical and material research fields, where higher spatial resolution, better sensitivity, deeper penetration and faster data acquisition are always desired. Most recent examples include fluorescence microscopy, coherent Raman and nonlinear wave mixing imaging. In this talk, we will present our recent progresses on deep tissue fluorescence super-resolution and non-labeling chiral sum frequency generation imaging by utilizing optical field engineering mechanism.
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