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Abstract Multifocal microscopy enables high-speed three-dimensional (3D) volume imaging by using a multifocal grating in the emission path. This grating is typically designed to afford a uniform illumination of multifocal subimages for a single emission wavelength. Using the same grating for multicolor imaging results in non-uniform subimage intensities in emission wavelengths for which the grating is not designed. This has restricted multifocal microscopy applications for samples having multicolored fluorophores. In this paper, we present a multicolor multifocal microscope implementation which uses a Spatial Light Modulator (SLM) as a single multifocal grating to realize near-uniform multifocal subimage intensities across multiple wavelength emission bands. Using real-time control of an in-situ-optimized SLM implemented as a multifocal grating, we demonstrate multicolor multifocal 3D imaging over three emission bands by imaging multicolored particles as well as Escherichia coli (E. coli) interacting with human liver cancer cells, at $$\sim 2.5$$ ∼ 2.5 multicolor 3D volumes per second acquisition speed. Our multicolor multifocal method is adaptable across SLM hardware, emission wavelength band locations and number of emission bands, making it particularly suited for researchers investigating fast processes occurring across a volume where multiple species are involved.

Proposed is a novel eye vision testing system based on micromachines that uses micro-optic, micromechanic, and microelectronic technologies. The micromachines include a programmable micro-optic lens and aperture control devices, pico-projectors, Radio Frequency (RF), optical wireless communication and control links, and energy harvesting and storage devices with remote wireless energy transfer capabilities. The portable lightweight system can measure eye refractive powers, optimize light conditions for the eye under testing, conduct color-blindness tests, and implement eye strain relief and eye muscle exercises via time sequenced imaging. A basic eye vision test system is built in the laboratory for near-sighted (myopic) vision spherical lens refractive error correction. Refractive error corrections from zero up to −5.0 Diopters and −2.0 Diopters are experimentally demonstrated using the Electronic-Lens (E-Lens) and aperture control methods, respectively. The proposed portable eye vision test system is suited for children’s eye tests and developing world eye centers where technical expertise may be limited. Design of a novel low-cost human vision corrective eyewear is also presented based on the proposed aperture control concept. Given its simplistic and economical design, significant impact can be created for humans with vision problems in the under-developed world.

Multifocal microscopy (MFM) offers high-speed three-dimensional imaging through the simultaneous image capture from multiple focal planes. Conventional MFM systems use a fabricated grating in the emission path for a single emission wavelength band and one set of focal plane separations. While a Spatial Light Modulator (SLM) can add more flexibility as a replacement to the fabricated grating, the relatively small number of pixels in the SLM chip, cross-talk between the pixels, and aberrations in the imaging system can produce non-uniform intensity in the different axially separated image planes. We present an in situ iterative SLM calibration algorithm that overcomes these optical- and hardware-related limitations to deliver near-uniform intensity across all focal planes. Using immobilized gold nanoparticles under darkfield illumination, we demonstrate superior intensity evenness compared to current methods. We also demonstrate applicability across emission wavelengths, axial plane separations, imaging modalities, SLM settings, and different SLM manufacturers. Therefore, our microscope design and algorithms provide an alternative to the use of fabricated gratings in MFM, as they are relatively simple and could find broad applications in the wider research community.