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Mechanobiology helps us to decipher cell and tissue functions by looking at changes in their mechanical properties that contribute to development, cell differentiation, physiology, and disease. Mechanobiology sits at the interface of biology, physics and engineering. One of the key technologies that enables characterization of properties of cells and tissue is microscopy. Combining microscopy with other quantitative measurement techniques such as optical tweezers and scissors, gives a very powerful tool for unraveling the intricacies of mechanobiology enabling measurement of forces, torques and displacements at play. We review the field of some light based studies of mechanobiology and optical detection of signal transduction ranging from optical micromanipulation─optical tweezers and scissors, advanced fluorescence techniques and optogenentics. In the current perspective paper, we concentrate our efforts on elucidating interesting measurements of forces, torques, positions, viscoelastic properties, and optogenetics inside and outside a cell attained when using structured light in combination with optical tweezers and scissors. We give perspective on the field concentrating on the use of structured light in imaging in combination with tweezers and scissors pointing out how novel developments in quantum imaging in combination with tweezers and scissors can bring to this fast growing field.

Abstract: Traumatic Brain Injury (TBI) arises from an external force affecting the brain, leading to a range of outcomes from mild to severe. Despite continuous scientific advancements, it continues to pose a persistent threat and remains a significant cause of physical impairment and mortality. Various models, including blast-induced TBI (bTBI), have been proposed to simulate TBI. Laser-induced shockwaves (LIS) us emerging as an effective method. LIS generates shockwaves via pulsed laser-induced plasma formation, offering a controlled means to study TBI at the cellular level. Astrocytes, pivotal in maintaining brain function post-injury, undergo dynamic morphological changes, contributing to the understanding of injury responses and neurodegenerative diseases. This study introduces a system combining Laser-Induced Shockwaves (LIS) and Quantitative Phase Microscopy (QPM) to quantify morphological changes in astrocytes during and after LIS exposure. QPM, a label-free method, facilitates 3D imaging and captures real-time cellular dynamics. The integration of LIS and QPM enables the assessment of astrocyte responses to shear stress caused by LIS, revealing immediate and sustained morphological transformations. Analysis post-LIS exposure indicates significant alterations in circularity, volume, surface area, and other features. Statistical tests affirm of observed trends, providing insights into astrocyte responses to mechanical forces. The findings contribute to understanding how mechanical stimuli impact astrocyte morphology, holding promise for targeted therapeutic strategies in traumatic brain injuries and related neurological disorders. The integrated LIS and QPM approach serves as a powerful tool for 3D imaging and quantitative measurement of astrocyte morphological changes, offering deeper insights into cellular dynamics and potential therapeutic interventions.

Optical tweezers are tools made of light that enable contactless pushing, trapping, and manipulation of objects, ranging from atoms to space light sails. Since the pioneering work by Arthur Ashkin in the 1970s, optical tweezers have evolved into sophisticated instruments and have been employed in a broad range of applications in the life sciences, physics, and engineering. These include accurate force and torque measurement at the femtonewton level, microrheology of complex fluids, single micro- and nano-particle spectroscopy, single-cell analysis, and statistical-physics experiments. This roadmap provides insights into current investigations involving optical forces and optical tweezers from their theoretical foundations to designs and setups. It also offers perspectives for applications to a wide range of research fields, from biophysics to space exploration.

Piezo1 belongs to mechano-activatable cation channels serving as biological force sensors. However, the molecular events downstream of Piezo1 activation remain unclear. In this study, we used biosensors based on fluorescence resonance energy transfer (FRET) to investigate the dynamic modes of Piezo1-mediated signaling and revealed a bimodal pattern of Piezo1-induced intracellular calcium signaling. Laser-induced shockwaves (LIS) and its associated shear stress can mechanically activate Piezo1 to induce transient intracellular calcium (Ca[i] ) elevation, accompanied by an increase in FAK activity. Interestingly, multiple pulses of shockwave stimulation caused a more sustained calcium increase and a decrease in FAK activity. Similarly, tuning the degree of Piezo1 activation by titrating either the dosage of Piezo1 ligand Yoda1 or the expression level of Piezo1 produced a similar bimodal pattern of FAK responses. Further investigations revealed that SHP2 serves as an intermediate regulator mediating this bimodal pattern in Piezo1 sensing and signaling. These results suggest that the degrees of Piezo1 activation induced by both mechanical LIS and chemical ligand stimulation may determine downstream signaling characteristics.

A major roadblock to the development of photonic sensors is the scattering associated with many biological systems. We show the conservation of photonic states through optically self-arranged biological waveguides, for the first time, which can be implemented to transmit light through scattering media. The conservation of optical properties of light through biological waveguides allows for the transmission of high bandwidth information with low loss through scattering media. Here, we experimentally demonstrate the conservation of polarization state and orbital angular momentum of light through a self-arranged biological waveguide, several centimeters long, in a sheep red blood cell suspension. We utilize nonlinear optical effects to self-trap cells, which form waveguides at 532 nm and 780 nm wavelengths. Moreover, we use the formed waveguide channels to couple and guide probe beams without altering the information. The formed biological waveguides are in a sub-diffusive scattering regime, so the photons' information degrades insignificantly over several centimeters of propagation through the scattering media. Our results show the potential of biological waveguides as a methodology for the development of novel photonic biosensors, biomedical devices that require optical wireless communication, and the development of new approaches to noninvasive biomedical imaging.

Laser induced shockwave (LIS) can be utilized to subject neuronal cells to conditions similar to those occurring during a blast induced traumatic brain injury. We utilized a 532nm Coherent Flare laser to induce a shockwave near cells which had been transfected with a FRET calcium biosensor (D3CPV) so that we could monitor the immediate cellular responses. Our shockwave system was characterized with a high-speed camera to monitor cavitation bubble dynamics and calculate the shear forces cells were subjected to. We found that we could induce forces which have been previously shown to induce injury. Using both phase and fluorescence microscopy we monitored the effects of shear on our cells. We found that at distances up to 120 microns from the laser focal point cells experienced shears greater than 10kPa. At those distances cell fragmentation was observed. Cells that survived and expressed the FRET biosensor demonstrated an immediate calcium elevation irrespective of extracellular or cytoplasmic calcium concentration. Cells recovered to pre-shockwave calcium levels within ∼30s. In conclusion, LIS can be utilized to simultaneously monitor the neuronal response to shear stress and nearby cell death or injury.

There is a need for new methodologies to investigate cell apoptosis and recovery, cell adhesion, and cell-cell interactions in cellular biology and neurobiology. Such systems should be able to induce localized cell injuries and measure damage responses from single cells. In this regard, pulsed lasers can be used to produce Laser- Induced Shockwaves (LIS), which can cause cell detachments and induce cellular membrane injuries, by applying shear force in order of μN. Furthermore, since the resulting shear force can increase membrane permeability, chemicals and markers can then be transferred into cells non-invasively. Continuous-wave lasers can be used as Optical Tweezers (OT), to apply non-contact delicate forces, as low as 0.1f N, and deliver materials into cells, and also move the cells to different locations. In this paper, we introduce a combination of modalities to apply variable forces, from femto to micro newtons, to cells. Our system consists of a 1060nm continuous laser light source for OT and a 1030nm femtosecond pulsed laser for generating LIS. To have a direct measurement of changes in the cellular thickness and membrane dynamics, the cells are imaged under a Quantitative Phase Microscope (QPM). Our microscope is capable of Differential-Interference Microscopy (DIC) and Phase-Contrast microscopy (PhC) and fluorescent microscopy, making it a unique system for studying cell injuries.

In this paper, we propose a new system for studying cellular injury. The system is a biophotonic work station that can generate Laser-Induced Shockwave (LIS) in the cell culture medium combined with a Quantitative Phase Microscope (QPM), enabling the real-time measurement of intracellular dynamics and quantitative changes in cellular thickness during the damage and recovery processes. In addition, the system is capable of Phase Contrast (PhC) and Differential Interference Contrast (DIC) microscopy. Our studies showed that QPM allows us to discern changes that otherwise would be unnoticeable or difficult to detect using phase or DIC imaging. As one application, this system enables the study of traumatic brain injury in vitro. Astrocytes are the most numerous cells in the central nervous system (CNS) and have been shown to play a role in the repair of damaged neuronal tissue. In this study, we use LIS to create a precise mechanical force in the culture medium at a controlled distance from astrocytes and measure the quantitative changes, in order of nanometers, in cell thickness. Experiments were performed in different cell culture media in order to evaluate the reproducibility of the experimental method.

Laser-induced shockwaves (LIS) can be utilized as a method to subject cells to conditions similar to those occurring during a blast-induced traumatic brain injury. The pairing of LIS with genetically encoded biosensors allows researchers to monitor the immediate molecular events resulting from such an injury. In this study, we utilized the genetically encoded Ca2+ FRET biosensor D3CPV to study the immediate Ca2+ response to laser-induced shockwave in cortical neurons and Schwann cells. Our results show that both cell types exhibit a transient Ca2+ increase irrespective of extracellular Ca2+ conditions. LIS allows for the simultaneous monitoring of the effects of shear stress on cells, as well as nearby cell damage and death.

Laser-induced shockwaves (LIS) can be utilized as a method to subject cells to conditions similar to those occurring during a blast-induced traumatic brain injury. The pairing of LIS with genetically encoded biosensors allows researchers to monitor the immediate molecular events resulting from such an injury. In this study, we utilized the genetically encoded Ca2+ FRET biosensor D3CPV to study the immediate Ca2+ response to laser-induced shockwave in cortical neurons and Schwann cells. Our results show that both cell types exhibit a transient Ca2+ increase irrespective of extracellular Ca2+ conditions. LIS allows for the simultaneous monitoring of the effects of shear stress on cells, as well as nearby cell damage and death.

Traumatic brain injury (TBI) occurs when an external shock causes injury to the brain. The mechanism of the disease is not completely understood yet. Studies have shown that astrocytes play various roles following brain injury. However, the exact functional role of them after TBI is still a matter of debate. Laser-induced shock waves (LIS) can create a precise controllable mechanical force that is capable of injuring or lysing cells to simulate the brain injury at the cellular level. Here, we propose a system that enables us to induce injuries in CNS cells with LIS and observe the whole process under a Quantitative phase microscope (QPM). Our system is also capable of adding another laser for optically trapping the cells to keep them at a certain distance from the center of the shockwave, as this distance is one of the important factors which determines the level of injury.

Since their inception, optical tweezers have proven to be a useful tool for improving human understanding of the microscopic world with wide-ranging applications across science. In recent years, they have found many particularly appealing applications in the field of biomedical engineering which harnesses the knowledge and skills in engineering to tackle problems in biology and medicine. Notably, metallic nanostructures like gold nanoparticles have proven to be an excellent tool for OT-based micromanipulation due to their large polarizability and relatively low cytotoxicity. In this article, we review the progress made in the application of optically trapped gold nanomaterials to problems in bioengineering. After an introduction to the basic methods of optical trapping, we give an overview of potential applications to bioengineering specifically: nano/biomaterials, microfluidics, drug delivery, biosensing, biophotonics and imaging, and mechanobiology/single-molecule biophysics. We highlight the recent research progress, discuss challenges, and provide possible future directions in this field.

Astrocytes respond to brain injury at a cellular level by the process of reactive astrogliosis, and are able to adjust their response according to the severity of the insult. Included in the reactive response is the process of phagocytosis, where astrocytes clean up surrounding cellular debris from damaged cells. In this study, we observe the process of phagocytosis by primary cortical astrocytes in the presence of media flow across the apical surface of the cells. Both static and cells under flow conditions respond consistently via phagocytosis of laser-induced cellular debris. We found that astrocytes exposed to shear flow initiate phagocytosis at a consistently faster rate than cells observed under static conditions. Shear forces created by laminar flow were analyzed as well as the flow fields created around astrocyte cells. Results suggest astrocyte phagocytosis is a mechanosensitive response, thus revealing the potential to enhance astrocyte phagocytic cleanup of damaged nervous tissue.

Understanding the mitotic DNA damage response (DDR) is critical to our comprehension of cancer, premature aging and developmental disorders which are marked by DNA repair deficiencies. In this study we use a micro-focused laser to induce DNA damage in selected mitotic chromosomes to study the subsequent repair response. Our findings demonstrate that (1) mitotic cells are capable of DNA repair as evidenced by DNA synthesis at damage sites, (2) Repair is attenuated when DNA-PKcs and ATM are simultaneously compromised, (3) Laser damage may permit the observation of previously undetected DDR proteins when damage is elicited by other methods in mitosis, and (4) Twenty five percent of mitotic DNA-damaged cells undergo a subsequent mitosis. Together these findings suggest that mitotic DDR is more complex than previously thought and may involve factors from multiple repair pathways that are better understood in interphase.

Understanding the mitotic DNA damage response (DDR) is critical to our comprehension of cancer, premature aging and developmental disorders which are marked by DNA repair deficiencies. In this study we use a micro-focused laser to induce DNA damage in selected mitotic chromosomes to study the subsequent repair response. Our findings demonstrate that (1) mitotic cells are capable of DNA repair as evidenced by DNA synthesis at damage sites, (2) Repair is attenuated when DNA-PKcs and ATM are simultaneously compromised, (3) Laser damage may permit the observation of previously undetected DDR proteins when damage is elicited by other methods in mitosis, and (4) Twenty five percent of mitotic DNA-damaged cells undergo a subsequent mitosis. Together these findings suggest that mitotic DDR is more complex than previously thought and may involve factors from multiple repair pathways that are better understood in interphase.

During cell-to-cell communications, the interplay between physical and biochemical cues is essential for informational exchange and functional coordination, especially in multicellular organisms. However, it remains a challenge to visualize intercellular signaling dynamics in single live cells. Here, we report a photonic approach, based on laser microscissors and Förster resonance energy transfer (FRET) microscopy, to study intercellular signaling transmission. First, using our high-throughput screening platform, we developed a highly sensitive FRET-based biosensor (SCAGE) for Src kinase, a key regulator of intercellular interactions and signaling cascades. Notably, SCAGE showed a more than 40-fold sensitivity enhancement than the original biosensor in live mammalian cells. Next, upon local severance of physical intercellular connections by femtosecond laser pulses, SCAGE enabled the visualization of a transient Src activation across neighboring cells. Lastly, we found that this observed transient Src activation following the loss of cell-cell contacts depends on the passive structural support of cytoskeleton but not on the active actomyosin contractility. Hence, by precisely introducing local physical perturbations and directly visualizing spatiotemporal transmission of ensuing signaling events, our integrated approach could be broadly applied to mimic and investigate the wounding process at single-cell resolutions. This integrated approach with highly sensitive FRET-based biosensors provides a unique system to advance our in-depth understanding of molecular mechanisms underlying the physical-biochemical basis of intercellular coupling and wounding processes.

Originally described in crane-fly spermatocytes, tethers physically link and transmit force between the ends of separating chromosomes. Optical tweezers and laser scissors were used to sever the tether between chromosomes, create chromosome fragments attached to the tether which move toward the opposite pole, and to trap the tethered fragments. Laser microsurgery in the intracellular space between separating telomeres reduced chromosome strain in half of tested chromosome pairs. When the telomere-containing region was severed from the rest of the chromosome body, the resultant fragment either traveled towards the proper pole (poleward), towards the sister pole (cross-polar), or movement ceased. Fragment travel towards the sister pole varied in distance and always ceased following a cut between telomeres, indicating the tether is responsible for transferring a cross-polar force to the fragment. Optical trapping of cross-polar traveling fragments places an upper boundary on the tethering force of ~1.5 pN.

Successful artificial insemination relies on the use of high quality spermatozoa. One measure of sperm quality is swimming force. Increased swimming force has been correlated with higher sperm swimming speeds and improved reproductive success. It is hypothesized that by increasing sperm swimming speed, one can increase swimming force. Previous studies have shown that red light irradiation causes an increase in sperm swimming speed. In the current study, 633nm red light irradiation is shown to increase mean squared displacement in trapped sperm. The methodology allows for comparison of relative swimming forces between irradiated and non-irradiated samples.

During the anaphase stage of mitosis, a motility force transports genetic material in the form of chromosomes to the poles of the cell. Chromosome deformations during anaphase transport have largely been attributed to viscous drag force, however LaFountain et. al. found that a physical tether connects separating chromosome ends in crane-fly spermatocytes such that a backwards tethering force elongates the separating chromosomes. In the presented study laser microsurgery was used to deduce the mechanistic basis of chromosome elongation in rat-kangaroo cells. In half of tested chromosome pairs, laser microsurgery between separating chromosome ends reduced elongation by 7±3% suggesting a source of chromosome strain independent of viscous drag. When microsurgery was used to sever chromosomes during transport, kinetochore attached fragments continued poleward travel while half of end fragments traveled towards the opposite pole and the remaining fragments either did not move or segregated to the proper pole. Microsurgery directed between chromosome ends always ceased cross-polar fragment travel suggesting the laser severed a physical tether transferring force to the fragment. Optical trapping of fragments moving towards the opposite pole estimates an upper boundary on the tethering force of 1.5 pN.

Traumatic Brain Injury (TBI) occurs when an external force injures the brain. While clinical outcomes of TBI can vary widely in severity, few mechanisms of neurodegeneration following TBI have been identified for treatment. We propose a model for studying TBI using laser-induced shockwaves (LISs). An optical system was developed that allows single cells to be studied in response to LISs. Our system utilizes an optically-coupled force measurement component that allows for the visualization of shockwave dynamics. Here, the force measurement system is characterized by imaging stages over the period of violent expansion and collapse of microbubbles responsible for shockwave generation.

The ability to successfully fertilize ova relies upon the swimming ability of spermatozoa. Both in humans and in animals, sperm motility has been used as a metric for the viability of semen samples. Recently, several studies have examined the efficacy of low dosage red light exposure for cellular repair and increasing sperm motility. Of prime importance to the practical application of this technique is the absence of DNA damage caused by radiation exposure. In this study, we examine the effect of 633 nm coherent, red laser light on sperm motility using a novel wavelet-based algorithm that allows for direct measurement of curvilinear velocity under red light illumination. This new algorithm gives results comparable to the standard computer-assisted sperm analysis (CASA) system. We then assess the safety of red light treatment of sperm by analyzing, (1) the levels of double-strand breaks in the DNA, and (2) oxidative damage in the sperm DNA. The results demonstrate that for the parameters used there are insignificant differences in oxidative DNA damage as a result of irradiation.

Red light has been shown to increase sperm swimming speeds, but there is little characterization of its effect on DNA or swimming force. In this study 633nm laser light irradiation did not induce significant levels of oxidative DNA damage.

Laser microsurgery directed between sister telomeres reduced strain in anaphase PtK2 chromosomes. Optical trapping of severed chromosome ends suggests this strain arises from a physical element which links sister chromatids during early anaphase.

The review describes the principles and examples of practical realization of diagnostic approaches based on the coherence theory, optical singularities and interference techniques. The presentation is based on the unified correlation-optics and coherence-theory concepts. The applications of general principles are demonstrated by several examples including the study of inhomogeneities and fluctuations in water solutions and methods for sensitive diagnostics of random phase objects (e.g., rough surfaces). The specific manifestations of the correlation-optics paradigms are illustrated in applications to non-monochromatic fields structured both in space and time. For such fields, the transient patterns of the internal energy flows (Poynting vector distribution) and transient states of polarization are described. The single-shot spectral interference is analyzed as a version of the correlation-optics approach adapted to ultra-short light pulses. As a characteristic example of such pulses, uniting the spatio-temporal and singular properties, the spatio-temporal optical vortices are considered in detail; their properties, methods of generation, diagnostics, and possible applications are exposed and characterized. Prospects of further research and applications are discussed. [ABSTRACT FROM AUTHOR]

Bacterial biofilms underlie many persistent infections, posing major hurdles in antibiotic treatment. Here we design and demonstrate 'tug-of-war' optical tweezers that can facilitate the assessment of cell-cell adhesion-a key contributing factor to biofilm development, thanks to the combined actions of optical scattering and gradient forces. With a customized optical landscape distinct from that of conventional tweezers, not only can such 'tug-of-war' tweezers stably trap and stretch a rod-shaped bacterium in the observing plane, but, more importantly, they can also impose a tunable lateral force that pulls apart cellular clusters without any tethering or mechanical movement. As a proof of principle, we examined a Sinorhizobium meliloti strain that forms robust biofilms and found that the strength of intercellular adhesion depends on the growth medium. This technique may herald new photonic tools for optical manipulation and biofilm study, as well as other biological applications.

A key goal in the conservation of endangered species is to increase successful reproduction. In cases where traditional methods of in vitro fertilization are unsuccessful, new methods of assisted reproduction are needed. One option is selective fertilization via optically trapped sperm. A more passive option is red light irradiation. Red light irradiation has been shown to increase sperm motility, thus increasing fertilizing potential. However, there is some concern that exposure to laser irradiation induces the production of oxidative species in cells, which can be damaging to DNA. In order to test the safety of irradiating sperm, sperm samples were exposed to 633 nm laser light and their DNA were tested for oxidative damage. Using fluorescence microscopy, antibody staining, and ELISA to detect oxidative DNA damage, it was concluded that red light irradiation does not pose a safety risk to sperm DNA. The use of red light on sperm has potential in both animal conservation and human reproduction techniques. This method can also be used in conjunction with optical trapping for viable sperm selection.

Bacterial biofilms underlie many persistent infections, posing major hurdles in antibiotic treatment. Here we design and demonstrate 'tug-of-war' optical tweezers that can facilitate the assessment of cell-cell adhesion-a key contributing factor to biofilm development, thanks to the combined actions of optical scattering and gradient forces. With a customized optical landscape distinct from that of conventional tweezers, not only can such 'tug-of-war' tweezers stably trap and stretch a rod-shaped bacterium in the observing plane, but, more importantly, they can also impose a tunable lateral force that pulls apart cellular clusters without any tethering or mechanical movement. As a proof of principle, we examined a Sinorhizobium meliloti strain that forms robust biofilms and found that the strength of intercellular adhesion depends on the growth medium. This technique may herald new photonic tools for optical manipulation and biofilm study, as well as other biological applications.

Optogenetics uses light to control and observe the activity of neurons, often using a focused laser beam. As brain tissue is a scattering medium, beams are distorted and spread with propagation through neural tissue, and the beam's degradation has important implications in optogenetic experiments. To address this, we present an analysis of scattering and loss of intensity of focused laser beams at different depths within the brains of zebrafish larvae. Our experimental set-up uses a 488 nm laser and a spatial light modulator to focus a diffraction-limited spot of light within the brain. We use a combination of experimental measurements of back-scattered light in live larvae and computational modelling of the scattering to determine the spatial distribution of light. Modelling is performed using the Monte Carlo method, supported by generalised Lorenz-Mie theory in the single-scattering approximation. Scattering in areas rich in cell bodies is compared to that of regions of neuropil to identify the distinct and dramatic contributions that cell nuclei make to scattering. We demonstrate the feasibility of illuminating individual neurons, even in nucleus-rich areas, at depths beyond 100 μm using a spatial light modulator in combination with a standard laser and microscope optics.

Cells within the body are subject to various forces; however, the details concerning the way in which cells respond to mechanical stimuli are not well understood. We demonstrate that laser-induced shockwaves (LIS) combined with biosensors based on fluorescence resonance energy transfer (FRET) is a promising new approach to study biological processes in single live cells. As "proof-of-concept," using a FRET biosensor, we show that in response to LIS, cells release intracellular calcium. With the parameters used, cells retain their morphology and remain viable. LIS combined with FRET permits observation of the cells immediate response to a sudden shear force.

Axonal injury and stress have long been thought to play a pathogenic role in a variety of neurodegenerative diseases. However, a model for studying single-cell axonal injury in mammalian cells and the processes of repair has not been established. The purpose of this study was to examine the response of neuronal growth cones to laser-induced axonal damage in cultures of embryonic rat hippocampal neurons and induced pluripotent stem cell (iPSC) derived human neurons. A 532-nm pulsed [Formula: see text] picosecond laser was focused to a diffraction limited spot at a precise location on an axon using a laser energy/power that did not rupture the cell membrane (subaxotomy). Subsequent time series images were taken to follow axonal recovery and growth cone dynamics. After laser subaxotomy, axons thinned at the damage site and initiated a dynamic cytoskeletal remodeling process to restore axonal thickness. The growth cone was observed to play a role in the repair process in both hippocampal and iPSC-derived neurons. Immunofluorescence staining confirmed structural tubulin damage and revealed initial phases of actin-based cytoskeletal remodeling at the damage site. The results of this study indicate that there is a repeatable and cross-species repair response of axons and growth cones after laser-induced damage.

We present a system which uses a single spatial light modulator to control the spin angular momentum of multiple optical traps. These traps may be independently controlled both in terms of spatial location and in terms of their spin angular momentum content. The system relies on a spatial light modulator used in a "split-screen" configuration to generate beams of orthogonal polarisation states which are subsequently combined at a polarising beam splitter. Defining the phase difference between the beams with the spatial light modulator enables control of the polarisation state of the light. We demonstrate the functionality of the system by controlling the rotation and orientation of birefringent vaterite crystals within holographic optical tweezers.

Light emerging from a spiral phase plate with a non-integer phase step has a complicated vortex structure and is unstable on propagation. We generate light carrying fractional orbital angular momentum (OAM) not with a phase step but by a synthesis of Laguerre-Gaussian modes. By limiting the number of different Gouy phases in the superposition we produce a light beam which is well characterised in terms of its propagation. We believe that their structural stability makes these beams ideal for quantum information processes utilising fractional OAM states.