![particle designer 2.5 particle designer 2.5](https://m.media-amazon.com/images/I/71Sb3PYhBgL._SX342_.jpg)
However, it was inevitable that ∼50% of the emitted fluorescence light was discarded because we used a liquid crystal on silicon (LCoS) spatial light modulator (SLM) to encode the wavefront, which inherently responds only to a certain state of linearly polarized light. Our 2.5DM showed ∼10-fold higher throughput and a >8-fold lower light dose compared to conventional high-throughput microscopy, which made it as a promising tool for single-molecule RNA imaging, immunofluorescence imaging, and long-term live-cell imaging. The fact that it is compatible with high numerical aperture (NA) objectives made it suitable for photon-limited applications such as single-molecule imaging. Unlike typical PSF engineering methods, such as a cubic phase mask or a tetrapod PSF for 3D single-molecule imaging, our approach allowed one to readily interpret original images without complicated image processing. The obtained PSF exhibited high uniformity of axial profiles and minimal side-lobes. Recently, we have demonstrated 2.5D microscopy (2.5DM) that substantially improved the throughput or imaging speed of fluorescence microscopy while maintaining high-resolution and single-molecule sensitivity by generating a defocus-invariant detection point spread function (PSF) within a specified depth-of-field (DOF). Nevertheless, these methods either demonstrated a low signal-to-noise ratio (SNR) or limited spatial resolution.
#Particle designer 2.5 serial
A variety of approaches have been proposed to achieve fast volumetric imaging using less or no serial z-scanning, including rapid focus-scanning using an electrically tunable lens, simultaneous 3D imaging by projecting multiple focal planes onto different areas of a camera and light field microscopy. However, the imaging speed of current high-throughput microscopy techniques is limited by the need for multiple z-scanning processes in order to obtain three-dimensional (3D) cell images. High-throughput and high-content fluorescence microscopy is an essential tool in cell biology and pharmacology as it enables us to study the activity and temporal dynamics of biomolecules with high spatial resolution in cells and tissues. © 2021 Optical Society of America under the terms of the OSA Open Access Publishing Agreement 1. We demonstrate the aberration correction capabilities and extended depth of field by imaging thick specimens with fewer z-scanning steps. We highlight the improved efficiency via quantitative single-molecule RNA imaging of mammalian cells with a 2-fold improvement in the fluorescence intensity compared to a conventional SLM-based microscopy. Here we report that a spatial light modulator (SLM) in a polarization insensitive configuration can significantly improve the detection efficiency of 2.5D microscopy, while also compensating for aberrations at large imaging depths caused by the refractive index mismatch between the sample and the immersion medium. Recently, single-shot 2.5D microscopy showed promising results for high-throughput quantitative subcellular analysis via extended depth of field imaging without sequential z-scanning however, the detection efficiency was limited and it lacked depth-induced aberration correction.
![particle designer 2.5 particle designer 2.5](https://media.springernature.com/full/springer-static/image/art%3A10.1038%2Fs41598-020-78854-6/MediaObjects/41598_2020_78854_Fig1_HTML.png)
For best results, use the separate Authors field to search for author names.
![particle designer 2.5 particle designer 2.5](https://i.ytimg.com/vi/tWjYHAlpJWE/maxresdefault.jpg)
Use quotation marks " " around specific phrases where you want the entire phrase only.Question mark (?) - Example: "gr?y" retrieves documents containing "grey" or "gray".Asterisk ( * ) - Example: "elect*" retrieves documents containing "electron," "electronic," and "electricity".Improve efficiency in your search by using wildcards.Example: (photons AND downconversion) - pump.
![particle designer 2.5 particle designer 2.5](https://s3.envato.com/files/265179649/Elemental-Inline-(Sale).png)
Example: (diode OR solid-state) AND laser.Note the Boolean sign must be in upper-case. Separate search groups with parentheses and Booleans.Keep it simple - don't use too many different parameters.