In clinical practice, a variety of diagnostic applications require the identification of target cells. Density has been used as a physical marker to distinguish cell populations since metabolic activities could alter the cell densities. Magnetic levitation offers great promise for separating cells at the single cell level within heterogeneous populations with respect to cell densities. Traditional magnetic levitation platforms need bulky and precise optical microscopes to visualize levitated cells. Moreover, the evaluation process of cell densities is cumbersome, which also requires trained personnel for operation. In this work, we introduce a device (HologLev) as a fusion of the magnetic levitation principle and lensless digital inline holographic microscopy (LDIHM). LDIHM provides ease of use by getting rid of bulky and expensive optics. By placing an imaging sensor just beneath the microcapillary channel without any lenses, recorded holograms are processed for determining cell densities through a fully automated digital image processing scheme. The device costs less than $100 and has a compact design that can fit into a pocket. We perform viability tests on the device by levitating three different cell lines (MDA-MB-231, U937, D1 ORL UVA) and comparing them against their dead correspondents. We also tested the differentiation of mouse osteoblastic (7F2) cells by monitoring characteristic variations in their density. Last, the response of MDA-MB-231 cancer cells to a chemotherapy drug was demonstrated in our platform. HologLev provides cost-effective, label-free, fully automated cell analysis in a compact design that could be highly desirable for laboratory and point-of-care testing applications.

Diamagnetic levitation is an emerging technology for remote manipulation of cells in cell and tissue level applications. Low-cost magnetic levitation configurations using permanent magnets are commonly composed of a culture chamber physically sandwiched between two block magnets that limit working volume and applicability. This work describes a single ring magnet-based magnetic levitation system to eliminate physical limitations for biofabrication. Developed configuration utilizes sample culture volume for construct size manipulation and long-term maintenance. Furthermore, our configuration enables convenient transfer of liquid or solid phases during the levitation. Before biofabrication, we first calibrated/ the platform for levitation with polymeric beads, considering the single cell density range of viable cells. By taking advantage of magnetic focusing and cellular self-assembly, millimeter-sized 3D structures were formed and maintained in the system allowing easy and on-site intervention in cell culture with an open operational space. We demonstrated that the levitation protocol could be adapted for levitation of various cell types (i.e., stem cell, adipocyte and cancer cell) representing cells of different densities by modifying the paramagnetic ion concentration that could be also reduced by manipulating the density of the medium. This technique allowed the manipulation and merging of separately formed 3D biological units, as well as the hybrid biofabrication with biopolymers. In conclusion, we believe that this platform will serve as an important tool in broad fields such as bottom-up tissue engineering, drug discovery and developmental biology.

Biomechanical changes at cellular level can dramatically affect living organisms in both aviation and space applications. Weightlessness induces morphological alteration of cells, which leads to tissue loss. Therefore, scientists have been studying the effect of weightlessness using cell culture based biological experiments using conventional microscopes. However, strict requirements regarding cost, weight and functionality limit the use of conventional microscopes in space environment. Lensless digital in-line holographic microscopy enables to use low-weight, low-cost and robust elements, such as a light emitting diode (LED), an aperture and an imaging sensor, instead of bulky, expensive and fragile optical elements, such as lenses, mirrors and filters. This technology offers a high field of view compared to conventional microscopes without affecting the resolution and it is also suitable for remote sensing applications with automated imaging capabilities. Here, we present a portable digital in-line holographic microscopy platform that allows to visualize cells and to analyze their viability in a microfluidic chip. The platform offers microscopic imaging with 1.55 μm spatial resolution, 21.7 mm 2 field of view and image coloring capability. This platform could potentially play an important role in space biotechnology applications by enabling low-cost, high-resolution and portable monitoring of cells.

Microparticle/cell separation is one of the most important applications in the field of biomedical sciences particularly for cell sorting and protein assays. There are variety of different separation technologies introduced in the literature that the main limitations are large amount of sample, expensive chemical use besides of requirement of a labeling procedure (i.e. fluorescent/magnetic labeling), complex machinery, and high operational costs. Magnetic levitationbased separation offers simple, rapid and precise separation of microparticles based on their densities by suspending them in a glass microcapillary between two opposing magnets. Traditionally, magnetic levitation-based microparticle separation and identification procedure is performed by imaging under bulky microscopes composed of fragile and expensive optics and require trained personnel to operate which makes the whole procedure costly, time consuming and prone to human error. Lensless digital inline holographic microscope (LDIHM) eliminates the need for sophisticated optics by replacing simple illumination and recording scheme that can be reduced into few widely-available and cost-effective components. Thus, inspection procedure is mostly carried out on digitally processing captured holograms so that dependency on optical components and human error is dramatically reduced alongside using cost-effective and handheld device. Here, we introduce a novel hybrid platform that brings the advantages of magnetic levitation system with lensless digital inline holographic microscope for precise separation and identification of microparticles based on their densities. In the platform, it was shown that 1.026 g/mL and 1.090 g/mL microparticles were successfully identified.

Separation of target cells in a heterogeneous solution is of great importance for clinical studies especially for immunology and oncology. Separated cells can be used for diagnostic applications ranging from whole blood counting to isolation of circulating tumor cells (CTC) for personalized medicine. Recent separation technologies rely on labelling and identifying target cells with variety of labelling principle such as fluorescence or magnetic tags. However, they require laborintensive processes, long analysis time, and expensive chemical reagents and instrumentation. Hence, their usage is limited to wellequipped centralized laboratories. There is a need for a rapid, sensitive, low-cost and automated cell separation technology to disseminate usage of this technology even in rural areas. Magnetic levitation is a powerful cell separation method, which distinguishes cells based on their levitation heights depending on cell density. However, magnetic levitation-based separation technologies require traditional, bulky and expensive microscopes for analysis. Lensless digital inline holographic microscopy (LDIHM) systems are composed of a simple illumination system containing an LED, a pinhole, and an imaging sensor for high-resolution microscopic imaging, which eliminates needs of highly fragile and expensive optics as in traditional microscopy. Here, we introduced a novel hybrid and portable cell separation platform, where magnetic levitation technology is integrated with LDIHM system for automated analysis of cell levitation heights. Using this platform, three different cell lines are successfully separated. Live and dead cells having distinguished levitation heights can be also identified in the platform.