High-resolution Imaging Techniques Research Articles

High resolution imaging of human development: shedding light on contrast agents.

Visualizing (micro)vascular structures remains challenging for researchers and clinicians due to limitations in traditional radiological imaging methods. Exploring the role of vascular development in craniofacial malformations in experimental settings can enhance understanding of these processes, with the effectiveness of high-resolution imaging techniques being crucial for successful research in this field. Micro-CT imaging offers 3D microstructural insights, but requires contrast-enhancing staining agents (CESAs) for visualizing (micro)-vascular tissues, known as contrast-enhanced micro-CT (CECT). As effective contrast agents are crucial for optimal visualization, this review focuses on comparative studies investigating such agents for micro-vascular tissue imaging using micro-CT. Furthermore, we demonstrate the utilization of B-Lugol solution as a promising contrast agent for acquiring high-quality micro-CT images of (micro)vascular structures in human embryonic samples. This scoping review followed Preferred Reporting Items for Systematic Reviews and Meta-analysis Protocols. PubMed database provided relevant articles, screened initially by title and abstract. Inclusion and exclusion criteria defined outcomes of interest. From an initial search, 273 records were identified, narrowed down to 9 articles after applying our criteria. Additionally, two articles were added through citation searching. This, a total of 11 articles were incorporated in this study. This micro-CT contrast agent review underscores the need for tailored choices based on research goals. Both Barium sulfate and Iodine-based agents showing excellent results, providing high resolution (micro) vascular content, especially in ex-vivo specimens. However, careful consideration of protocols and tissue characteristics remains imperative for optimizing the effectiveness of micro-CT imaging for the study of cranio-facial vascular development.

OCTA: Essential or Gimmick?

This commentary article delves into the transformative role of optical coherence tomography angiography (OCTA) in diagnosing and managing a wide array of eye conditions, including diabetic retinopathy, age-related macular degeneration, retinal vein occlusions, and white dot syndromes. Developed in 2005, OCTA has emerged as a non-invasive, high-resolution imaging technique that offers advantages over traditional fluorescein angiography (FA), providing quicker and safer monitoring of ocular conditions with similar diagnostic accuracy. In diabetic retinopathy, OCTA has been instrumental in early identification of retinal changes, offering quantifiable metrics including perfused capillary density (PCD) for assessing vascular alterations. For age-related macular degeneration (AMD), OCTA has deepened our understanding of non-exudative neovascular AMD, allowing for more effective monitoring and potential earlier initiation of treatment. In cases of retinal vein occlusions, OCTA can reveal specific microvascular features and allow for depth-resolved measurements of the foveal avascular zone, providing significant prognostic implications. OCTA has also been invaluable in studying rare white dot syndromes, enabling nuanced differentiation between conditions that often present similarly. Emerging research also suggests that OCTA can have potential utility in neurodegenerative diseases like Alzheimer's, where retinal vascular patterns could offer diagnostic insights. While OCTA is revolutionizing ophthalmic care, further clinical trials and standardization are needed for its broader adoption into clinical practice.

Diagnostic value of high-resolution vessel wall imaging technique in intracranial arterial stenosis and occlusion: a comparative analysis with digital subtraction angiography

Objective To analyze the diagnostic value of HR-VWI in intracranial arterial stenosis and occlusion and compare it with DSA. Methods A retrospective analysis of clinical data of 59 patients with intracranial arterial stenosis in our hospital was conducted to compare the diagnostic results of the two methods for different degrees of intracranial stenosis and various morphological plaques. Results The diagnosis of stenosis and occlusion by both methods showed no significant difference (p > 0.05). Comparison of plaque morphology detected by HR-VWI with pathological examination results showed no significant difference (p > 0.05); however, there was a significant difference between plaque morphology detected by DSA and pathological examination results (p < 0.05). Additionally, there was a significant difference between plaque morphology detected by HR-VWI and DSA (p < 0.05). Conclusion HR-VWI technique is comparable to DSA technique in diagnosing intracranial arterial stenosis and occlusion, but it is superior to DSA in plaque morphology diagnosis.

Exploring the Capability of Cu-MoS2 Catalysts for Use in Electrocatalytic Overall Water Splitting

Herein, we prepare MoS2 and Cu-MoS2 catalysts using the solvothermal method, a widely accepted technique for electrocatalytic overall water-splitting applications. TEM and SEM images, standard tools in materials science, provide a clear view of the morphology of Cu-MoS2. HRTEM analysis, a high-resolution imaging technique, confirms the lattice spacing, lattice plane, and crystal structure of Cu-MoS2. HAADF and corresponding color mapping and advanced imaging techniques reveal the existence of the Cu-doping, Mo, and S elements in Cu-MoS2. Notably, Cu plays a crucial role in improving the hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) of the Cu-MoS2 catalyst as compared with the MoS2 catalyst. In addition, the Cu-MoS2 catalyst demonstrates significantly lower overpotential (167.7 mV and 290 mV) and Tafel slopes (121.5 mV dec−1 and 101.5 mV dec−1), standing at −10 mA cm−2 and 10 mA cm−2 for HER and OER, respectively, compared to the MoS2 catalyst. Additionally, the Cu-MoS2 catalyst displays outstanding stability for 12 h at −10 mA cm−2 of HER and 12 h at 10 mA cm−2 of OER using chronopotentiaometry. Interestingly, the Cu-MoS2‖Cu-MoS2 cell displays a lower cell potential of 1.69 V compared with the MoS2‖MoS2 cell of 1.81 V during overall water splitting. Moreover, the Cu-MoS2‖Cu-MoS2 cell shows excellent stability when using chronopotentiaometry for 18 h at 10 mA cm−2.

EDTP enhances and protects the fluorescent signal of GFP in cleared and expanded tissues

Advanced 3D high-resolution imaging techniques are essential for investigating biological challenges, such as neural circuit analysis and tumor microenvironment in intact tissues. However, the fluorescence signal emitted by endogenous fluorescent proteins in cleared or expanded biological samples gradually diminishes with repeated irradiation and prolonged imaging, compromising its ability to accurately depict the underlying scientific problem. We have developed a strategy to preserve fluorescence in cleared and expanded tissue samples during prolonged high-resolution three-dimensional imaging. We evaluated various compounds at different concentrations to determine their ability to enhance fluorescence intensity and resistance to photobleaching while maintaining the structural integrity of the tissue. Specifically, we investigated the impact of EDTP utilization on GFP, as it has been observed to significantly improve fluorescence intensity, resistance to photobleaching, and maintain fluorescence during extended room temperature storage. This breakthrough will facilitate extended hydrophilic and hydrogel-based clearing and expansion methods for achieving long-term high-resolution 3D imaging of cleared biological tissues by effectively safeguarding fluorescent proteins within the tissue.

Dual-beam X-ray nano-holotomography.

Nanotomography with hard X-rays is a widely used technique for high-resolution imaging, providing insights into the structure and composition of various materials. In recent years, tomographic approaches based on simultaneous illuminations of the same sample region from different angles by multiple beams have been developed at micrometre image resolution. Transferring these techniques to the nanoscale is challenging due to the loss in photon flux by focusing the X-ray beam. We present an approach for multi-beam nanotomography using a dual-beam Fresnel zone plate (dFZP) in a near-field holography setup. The dFZP generates two nano-focused beams that overlap in the sample plane, enabling the simultaneous acquisition of two projections from slightly different angles. This first proof-of-principle implementation of the dual-beam setup allows for the efficient removal of ring artifacts and noise using machine-learning approaches. The results open new possibilities for full-field multi-beam nanotomography and pave the way for future advancements in fast holotomography and artifact-reduction techniques.

Feasibility of detecting shadows in disks induced by infall

Observations performed with high-resolution imaging techniques have revealed the existence of shadows in circumstellar disks that can be explained by the misalignment of an inner disk with respect to an outer one. The cause of misalignment, however, is still a matter of debate. In this study, we investigate the feasibility of observing shadows induced by one prominent scenario that may lead to misalignment, which involves the late infall of material onto a protostellar system. In particular, we used previously performed hydrodynamical simulations of such events and we generated flux maps in the visible, near-infrared, submillimeter, and millimeter wavelength ranges using Monte Carlo radiative transfer. Based on those results, we derived synthetic observations of these systems performed with the instruments SPHERE/VLT and ALMA, which we used as a basis for our subsequent analysis. We find that near-infrared observations with SPHERE are particularly well suited for detecting shadows via direct imaging alongside other features such as gaps, arcs, and streamers. On the contrary, performing a shadow detection based on reconstructed ALMA observations is very challenging due to the high sensitivity that is required for this task. Thus, in cases that allow for a detection, sophisticated analyses may be needed, for instance by the utilization of carefully constructed azimuthal profiles, aiding the search for potentially shallow shadows. Lastly, we conclude that a late infall-induced disk misalignment offers a plausible explanation for the emergence of shadows observed in various systems.

CFZA camera: a high-resolution lensless imaging technique based on compound Fresnel zone aperture.

In lensless imaging using a Fresnel zone aperture (FZA), it is generally believed that the resolution is limited by the outermost ring breadth of the FZA. The limitation has the potential to be broken according to the multi-order property of binary FZAs. In this Letter, we propose to use a high-order component of the FZA as the point spread function (PSF) to develop a high-order transfer function backpropagation (HBP) algorithm to enhance the resolution. The proportion of high-order diffraction energy is low, leading to severe defocus noise in the reconstructed image. To address this issue, we propose a Compound FZA (CFZA), which merges two partial FZAs operating at different orders as the mask to strike a balance between the noise and resolution. Experimental results verify that the CFZA-based camera has a resolution that is double that of a traditional FZA-based camera with an identical outer ring breadth and can be reconstructed with high quality by a single HBP without calibration. Our method offers a cost-effective solution for achieving high-resolution imaging, expanding the potential applications of FZA-based lensless imaging in a variety of areas.

The Pivotal Role of Microscopy in Unravelling the Nature of Microbial Deterioration of Waterlogged Wood: A Review

This review focuses on the pivotal role microscopy has played in diagnosing the type(s) of microbial attacks present in waterlogged ancient wooden objects, and to understand the nature and extent of deterioration of such objects. The microscopic journey began with the application of light microscopy (LM) to examine the deterioration of waterlogged woods, notably foundation piles supporting historic buildings, progressing into the use of high-resolution imaging tools (SEM and TEM) and techniques. Although bacteria were implicated in the deterioration of foundation piles, confirmation that bacteria can indeed degrade wood in its native state came when decaying wood from natural environments was examined using electron microscopy, particularly TEM, which enabled bacterial association with cell wall regions undergoing degradation to be clearly resolved. The information base has been a catalyst, stimulating numerous studies in the past three decades or so to understand the nature of microbial degradation of waterlogged archaeological wood more precisely, combining LM, SEM, and TEM with high-resolution chemical analytical methods, including chemical microscopy. The emerging information is aiding targeted developments towards a more effective conservation of ancient wooden objects as they begin to be uncovered from burial and waterlogging environments.

Three short-period Earth-sized planets around M dwarfs discovered by TESS: TOI-5720 b, TOI-6008 b, and TOI-6086 b

One of the main goals of the NASA Transiting Exoplanet Survey Satellite (TESS) mission is the discovery of Earth-like planets around nearby M-dwarf stars. We present the discovery and validation of three new short-period Earth-sized planets orbiting nearby M dwarfs: TOI-5720\,b, TOI-6008\,b, and TOI-6086\,b. We combined TESS data, ground-based multicolor light curves, ground-based optical and near-infrared spectroscopy, and Subaru/IRD radial velocity data to validate the planetary candidates and constrain the physical parameters of the systems. In addition, we used archival images, high-resolution imaging, and statistical validation techniques to support the planetary validation. TOI-5720\,b is an Earth-sized planet with a radius of $R_p = 1.09 0.07 R_ It orbits a nearby (36 pc) M2.5 host with an orbital period of $P = 1.4344555 0.0000036 $ days. It has an equilibrium temperature of eq 19$ K (assuming a null albedo) and an incident flux of $S_p = 41.7 4.5 S_ TOI-6008\,b is a short-period planet of $ P = 0.8574347 0.0000424$ day. It has a radius of $R_p = 1.03 0.05 R_ an equilibrium temperature of eq 19$ K, and an incident flux of $S_p = 41.5 4.5 S_ The host star (TOI-6008) is a nearby (23 pc) M5 with an effective temperature of eff 75 K$. Based on the radial velocity measurements collected with Subaru/IRD, we set a $3 upper limit of $M_p &lt; 4M_ thus ruling out a star or brown dwarf as the transiting companion. TOI-6086\,b orbits its nearby (32 pc) M3 host star eff 75$ K) every $1.3888725 0.0000827$ days and has a radius of $ R_p = 1.18 0.07 R_ an equilibrium temperature of eq 16$ K, and an incident flux of $S_p = 26.8 2.7 S_ Additional high-precision radial velocity measurements are needed to derive the planetary masses and bulk densities and to search for additional planets in the systems. Moreover, short-period Earth-sized planets orbiting around nearby M dwarfs are suitable targets for an atmospheric characterization with the James Webb Space Telescope through transmission and emission spectroscopy and phase-curve photometry.

Acute complicated jejunum diverticulitis: a case report with a short literature review.

Jejunal diverticulosis is a rare condition. Most of the time, it is asymptomatic; but it can cause severe complications such as intestinal perforation, mechanical occlusion, and hemorrhage. A patient aged 78 years, with a history of biological aortic valve prosthesis, atrial fibrillation, type 2 diabetes mellitus, and chronic obstructive pulmonary disease, presented in the emergency department for acute abdominal pain in the lower abdominal floor, nausea, and inappetence. Abdominal computed tomography revealed an inflammatory block in the hypogastrium, agglutinated small intestinal loops, fecal stasis, and air inclusions. Pulled mesentery and associated internal hernia are suspected. Exploratory laparotomy was performed, revealing an inflammatory block in the hypogastrium, whose dissection revealed inner purulent collection and the appearance of jejunal diverticulitis, a diagnosis confirmed by histopathological examination. Segmental resection of the jejunum with double-layer terminal-terminal enteroenteric anastomosis, lavage, and drainage was performed. The evolution was favorable. Based on our brief review, the diagnosis of complicated jejunal diverticulosis is difficult and sometimes not accurately established, even by high-resolution imaging techniques, with diagnostic laparotomy being necessary for these situations. Surgical treatment should be considered before severe complications develop.

All-dielectric super-lattice metasurfaces with fivefold spatial resolution enhancement for structured illumination microscopy.

Structured illumination microscopy (SIM) is a promising imaging technique for high-resolution imaging with a wide field of view. Although a periodic nanostructure is a versatile platform for engineering the spatial frequency of structured illumination patterns in SIM, challenges remain, including artifacts from Fourier space gaps. We designed an all-dielectric super-lattice metasurface (ADSLM) to generate structured illumination patterns with enhanced spatial frequency and broadened spatial frequency coverage with no intermediate frequency gaps. Our numerical simulations reveal that ADSLM-based image reconstruction is capable of producing high-contrast, artifact-free images, resulting in enhanced spatial resolution up to 5.7-fold for coherent SIM at 450 nm. Our results show that the ADSLM-SIM technique may facilitate high-resolution imaging using CMOS-compatible substrates, offering potential for compact miniaturized imaging applications.

Infrared imaging with nonlinear silicon resonator governed by high-Q quasi-BIC states

Nonlinear light-matter interactions have emerged as a promising platform for various applications, including imaging, nanolasing, background-free sensing, etc. Subwavelength dielectric resonators offer unique opportunities for manipulating light at the nanoscale and miniturising optical elements. Here, we explore the resonantly enhanced four-wave mixing (FWM) process from individual silicon resonators and propose an innovative FWM-enabled infrared imaging technique that leverages the capabilities of these subwavelength resonators. Specifically, we designed high-Q silicon resonators hosting dual quasi-bound states in the continuum at both the input pump and signal beams, enabling efficient conversion of infrared light to visible radiation. Moreover, by employing a point-scanning imaging technique, we achieve infrared imaging conversion while minimising the dependence on high-power input sources. This combination of resonant enhancement and point-scanning imaging opens up new possibilities for nonlinear imaging using individual resonators and shows potential in advancing infrared imaging techniques for high-resolution imaging, sensing, and optical communications.

Arbitrary engineering of spatial caustics with 3D-printed metasurfaces

Caustics occur in diverse physical systems, spanning the nano-scale in electron microscopy to astronomical-scale in gravitational lensing. As envelopes of rays, optical caustics result in sharp edges or extended networks. Caustics in structured light, characterized by complex-amplitude distributions, have innovated numerous applications including particle manipulation, high-resolution imaging techniques, and optical communication. However, these applications have encountered limitations due to a major challenge in engineering caustic fields with customizable propagation trajectories and in-plane intensity profiles. Here, we introduce the “compensation phase” via 3D-printed metasurfaces to shape caustic fields with curved trajectories in free space. The in-plane caustic patterns can be preserved or morphed from one structure to another during propagation. Large-scale fabrication of these metasurfaces is enabled by the fast-prototyping and cost-effective two-photon polymerization lithography. Our optical elements with the ultra-thin profile and sub-millimeter extension offer a compact solution to generating caustic structured light for beam shaping, high-resolution microscopy, and light-matter-interaction studies.

Overexpression of PEX14 results in mistargeting to mitochondria, accompanied by organelle fragmentation and clustering in human embryonic kidney cells

Peroxisome biogenesis disorders are caused by pathogenic variants in genes involved in biogenesis and maintenance of peroxisomes. However, mitochondria are also often affected in these diseases. Peroxisomal membrane proteins, including PEX14, have been found to mislocalise to mitochondria in cells lacking peroxisomes. Recent studies indicated that this mislocalisation contributes to mitochondrial abnormalities in PEX3-deficient patient fibroblasts cells. Here, we studied whether mitochondrial morphology is also affected in PEX3-deficient HEK293 cells and whether PEX14 mislocalises to mitochondria in these cells. Using high-resolution imaging techniques, we show that although endogenous PEX14 mislocalises to mitochondria, mitochondrial morphology was normal in PEX3-KO HEK293 cells. However, we discovered that overexpression of tagged PEX14 in wild-type HEK293 cells resulted in its mitochondrial localisation, accompanied by altered mitochondrial morphology. Our data indicate that overexpression of tagged PEX14 alone directly or indirectly cause mitochondrial abnormalities in cells containing peroxisomes.

Gluteal femoral subcutaneous and dermal adipose tissue in female.

During the sexual maturation, gluteal femoral adipose tissue is subjected to numerous modifications, not observable in other regions, in particular in women and less in men. Other authors described this region, but they used imaging techniques having lower resolution, than MRI proposed in this study. High resolution imaging techniques might provide important and more detailed information about the anatomy of gluteal femoral region. This study has been performed using 7 T-magnetic resonance imaging and ultrastructural analysis in order to provide accurate description of the subcutaneous adipose tissue and dermis of gluteal femoral region. In this study specimens harvested from cadavers and form living patients have been analyzed. The results showed the presence of three layers: superficial, middle, and deep, characterized by different organization of fat lobules. High resolution imaging showed the adipose papilla that originates from dermis and protrude in subcutaneous adipose tissue. Adipose papilla is characterized by a peculiar morphology with a basement, a neck and a head and these elements represent the functional subunits of adipose papilla. Moreover, ultrastructural study evidenced the relationship between adipocytes and sweat glands, regulated by lipid vesicles. This study provides important information about subcutaneous and dermal fat anatomy of gluteal femoral region, improving the past knowledge, and move toward a better understanding of the cellulite physiopathology.

X-Ray-Based 3D Histopathology of the Kidney Using Cryogenic Contrast-Enhanced MicroCT.

The kidney's microstructure, which comprises a highly convoluted tubular and vascular network, can only be partially revealed using classical 2D histology. Considering that the kidney's microstructure is closely related to its function and is often affected by pathologies, there is a need for powerful and high-resolution 3D imaging techniques to visualize the microstructure. Here, we present how cryogenic contrast-enhanced microCT (cryo-CECT) allowed 3D visualization of glomeruli, tubuli, and vasculature. By comparing different contrast-enhancing staining agents and freezing protocols, we found that the preferred sample preparation protocol was the combination of staining with 1:2 hafnium(IV)-substituted Wells-Dawson polyoxometalate and freezing by submersion in isopentane at -78°C. This optimized protocol showed to be highly sensitive, allowing to detect small pathology-induced microstructural changes in a mouse model of mild trauma-related acute kidney injury after thorax trauma and hemorrhagic shock. In summary, we demonstrated that cryo-CECT is an effective 3D histopathological tool that allows to enhance our understanding of kidney tissue microstructure and their related function.

Combining Reflectance Confocal Microscopy, Optical Coherence Tomography and Ex-Vivo Fluorescence Confocal Microscopy for Margin Assessment in Basal Cell Carcinoma Excision.

Recent developments of noninvasive, high-resolution imaging techniques, such as reflectance confocal microscopy (RCM) and optical coherence tomography (OCT), have enhanced skin cancer detection and precise tumor excision particularly in highly aggressive and poorly defined basal cell carcinomas (BCCs). The aim of this pilot study is to assess the feasibility and reproducibility of a systematic clinical workflow combining noninvasive (RCM-OCT) and invasive fluorescence confocal microscopy (FCM) imaging modalities in pre- and intra-surgical evaluations of the lateral and deep margins of BCC. Superficial incisions were made 2 mm beyond the clinical-dermoscopic BCC margins. Lateral margins were then explored with OCT and RCM. In positive margins, a further cut was made 2 mm distal from the previous. A final RCM/OCT-based double-negative margin was drawn around the entire perimeter of the lesion before referring to surgery. The freshly excised specimen was then examined with FCM (ex-vivo) for the evaluation of the deep margin. Histopathologic examination eventually confirmed margin involvement. The study included 22 lesions from 13 patients. At the end of the study, 146 margins-106 negative (73%) and 40 positive (27%) at RCM/OCT-were collected. The RCM/OCT margin evaluation showed an overall sensitivity of 100% and a specificity of 96.3%. The overall positive margins diagnostic accuracy was 98.2%. Reproducibility was evaluated on recorded images and the raters showed a substantial inter-observer agreement on both RCM (κ = 0.752) and OCT images (κ = 0.724). The combined RCM/OCT/FCM ex-vivo approach noninvasively facilitates the presurgical and intrasurgical lateral and deep margin assessment of poorly defined BCCs.

High-resolution Diffusion-weighted Imaging to Detect Changes in Tumor Size and ADC, and Predict Adverse Biopsy Histology during Prostate Cancer Active Surveillance.

Majority of men with low-risk prostate cancer can be managed with active surveillance (AS). This study evaluates a high-resolution diffusion-weighted imaging (HR-DWI) technique to predict adverse biopsy histology (AH), defined as Gleason score ≥7 on any biopsy or ≥3 increase in number of positive biopsy cores on systematic biopsies. We test the hypothesis that high-grade disease and progressing disease undergo subtle changes during even short intervals that can be detected by HR-DWI. In a prospective clinical trial, serial multiparametric MRIs, incorporating HR-DWI and standard DWI (S-DWI) were performed approximately 12 months apart prior to prostate biopsy (n = 59). HR-DWI, which uses reduced field-of-view and motion compensation techniques, was compared with S-DWI. HR-DWI had a 3-fold improvement in spacial resolution compared with S-DWI as confirmed using imaging phantoms. For detecting AH, multiparametric MRI using HR-DWI had a sensitivity of 75% and specificity of 83.9%, and MRI using S-DWI had a sensitivity of 71.4% and specificity of 54.8%. The AUC for HR-DWI was significantly higher (0.794 vs. 0.631, P = 0.014). Secondary analyses of univariable predictors of AH showed tumor size increase [OR 16.8; 95% confidence interval (CI): 4.06-69.48; P < 0.001] and apparent diffusion coefficient (ADC) decrease (OR 5.06; 95% CI: 1.39-18.38; P = 0.014) on HR-DWI were significant predictors of AH. HR-DWI outperforms S-DWI in predicting AH. Patient with AH have tumors that change in size and ADC that could be detected using HR-DWI. Future studies with longer follow-up should assess HR-DWI for predicting disease progression during AS. We report on a prospective clinical trial using a MRI that has three times the resolution of standard MRI. During AS for prostate cancer, two high-resolution MRIs performed approximately a year apart can detect tumor changes that predict the presence of aggressive cancers that should be considered for curative therapy such as prostatectomy or radiation.

Astigmatism-based active focus stabilisation with universal objective lens compatibility, extended operating range and nanometer precision

Focus stabilisation is vital for long-term fluorescence imaging, particularly in the case of high-resolution imaging techniques. Current stabilisation solutions either rely on fiducial markers that can be perturbative, or on beam reflection monitoring that is limited to high-numerical aperture objective lenses, making multimodal and large-scale imaging challenging. We introduce a beam-based method that relies on astigmatism, which offers advantages in terms of precision and the range over which focus stabilisation is effective. This approach is shown to be compatible with a wide range of objective lenses (10x-100x), typically achieving &lt;10 nm precision with &gt;10 μm operating range. Notably, our technique is largely unaffected by pointing stability errors, which in combination with implementation through a standalone Raspberry Pi architecture, offers a versatile focus stabilisation unit that can be added onto most existing microscope setups.