In this investigation, we propose a strip segmentation phase (SSP) method for a spatial light modulator (SLM) to generate independent multifocal spots when the beam passes through a high numerical aperture (NA) lens. With the SSP method, multifocal spots can be generated with each spot independently, flexibly and uniformly distributed. The performance of the SSP method is first validated with numerical simulation. Then, by applying the modulation method with SLM and importing the beams into an inverted fluorescence microscopy system with a high-NA lens, the spot distribution and their shapes can be observed by fluorescent image. The fluorescent image exhibits high uniformity and high consistency with the aforementioned numerical simulations. Finally, we dynamically load a series of phase maps on SLM to realize continuous and independent spot movement in a multifocal array. By laser direct writing on photoresist, a complex NWU-shape structure can be realized flexibly with multi-task fabrication capability. The SSP method can significantly improve the efficiency and flexibility of laser direct writing. It is also compatible with most recent techniques, e.g., multiphoton absorption, stimulated emission depletion and photo-induced depolymerization etc., to realize parallel super-resolution imaging and fabrications.

Flow cytometry is a widespread and powerful technique whose resolution is determined by its capacity to accurately distinguish fluorescently positive populations from negative ones. However, most informative results are discarded while performing the measurements of conventional flow cytometry, e.g., the cell size, shape, morphology, and distribution or location of labeled exosomes within the unpurified biological samples. Herein, we propose a novel approach using an anti-diffraction light sheet with anisotroic feature to excite fluorescent tags. Constituted by an anti-diffraction Bessel–Gaussian beam array, the light sheet is 12 μm wide, 12 μm high, and has a thickness of ~0.8 μm. The intensity profile of the excited fluorescent signal can, therefore, reflect the size and allow samples in the range from O (100 nm) to 10 μm (e.g., blood cells) to be transported via hydrodynamic focusing in a microfluidic chip. The sampling rate is 500 kHz, which provides a capability of high throughput without sacrificing the spatial resolution. Consequently, the proposed anti-diffraction light sheet flow cytometry (ADLSFC) can obtain more informative results than the conventional methodologies, and is able to provide multiple characteristics (e.g., the size and distribution of fluorescent signal) helping to distinguish the target samples from the complex backgrounds.