The scope of investigation is also different for additively manufactured optics. Novel characterization methods for printed optics are needed because traditional, destructive methods often do not work on these optics. Since additive manufacturing has become increasingly popular in prototyping, printed optics are also beginning to enter the market. Then, the application on living adherent and suspended cells is illustrated. The technique is firstly characterized by investigations on microspheres. Image separation is achieved by Fourier-transformation-based numerical demultiplexing procedures. The technique is based on spatially multiplexed interferometric microscopy (SMIM), where off-axis holograms are generated by spatial multiplexing of the illuminated sample plane utilizing a diffraction grating, which is extended by additional superposition of complementary white-light image information. Here, we investigate a single-snapshot multimodal approach that enables simultaneous acquisition of DHM-based QPI and BF images and is compatible with common optical microscopes for the analysis of living cells. However, such arrangements can be cost-intensive and/or limited in speed for imaging of dynamic processes. Multimodal microscopy is often achieved by sequential acquisition of information from different imaging modalities or simultaneously by capturing images with different acquisition devices. DHM can be implemented with other optical microscopy techniques like Bright Field (BF) or fluorescence, for example, to provide complementary information about cellular processes. DHM provides Quantitative Phase Imaging (QPI) of the light transmitted or scattered by an object, which allows to extract information about morphology and dynamics of investigated specimens. We also show that MAGNIFY provides a novel, accessible tool for improving the precision, utility, and generality of nanoscopy.ĭigital Holographic Microscopy (DHM) is an emerging label-free modality for quantitative imaging of semi-transparent biological specimens. Here, we demonstrate that MAGNIFY provides a generalized solution for imaging nanoscale subcellular features of a broad range of biological specimens. This facilitates nanoscale imaging (~25-nm effective resolution) using an ∼280-nm diffraction-limited objective lens on a conventional optical microscope and can be furthered to ~15 nm effective resolution if combined with computational methods such as Super-resolution Optical Fluctuation Imaging (SOFI). By using a mechanically sturdy hydrogel, MAGNIFY is capable of expanding biological specimens up to 11×. We present a new ExM framework, Molecule Anchorable Gel-enabled Nanoscale In-situ Fluorescence MicroscopY (MAGNIFY), that exhibits a broad retention of nucleic acids, proteins, and lipids without the need for a separate anchoring step. In addition, these protocols rely on mechanically fragile hydrogels that only expand by at most 4.5× linearly. However, most reported techniques are unable to preserve endogenous epitopes due to strong protease digestion used to expand samples. It achieves this by physically and isotropically magnifying preserved biological specimens embedded in a cross-linked water-swellable hydrogel. Expansion microscopy (ExM) is a powerful imaging strategy that offers a low-cost solution for interrogating biological systems at the nanoscale using conventional optical microscopes.
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