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Magnus Andersson

Publications and source records attributed to Magnus Andersson.

At least 19 recordsLinked to original sources

Segmentation and Characterization of Macerated Fibers and Vessels Using Deep Learning

Wood comprises different cell types, such as fibers, tracheids and vessels, defining its properties. Studying cells' shape, size, and arrangement in microscopy images is crucial for understanding wood characteristics. Typically, this involves macerating (soaking) samples in a solution to separate cells, then spreading them on slides for imaging with a microscope that covers a wide area, capturing thousands of cells. However, these cells often cluster and overlap in images, making the segmentation difficult and time-consuming using standard image-processing methods. In this work, we developed an automatic deep learning segmentation approach that utilizes the one-stage YOLOv8 model for fast and accurate segmentation and characterization of macerated fiber and vessel form aspen trees in microscopy images. The model can analyze 32,640 x 25,920 pixels images and demonstrate effective cell detection and segmentation, achieving a mAP_{0.5-0.95} of 78 %. To assess the model's robustness, we examined fibers from a genetically modified tree line known for longer fibers. The outcomes were comparable to previous manual measurements. Additionally, we created a user-friendly web application for image analysis and provided the code for use on Google Colab. By leveraging YOLOv8's advances, this work provides a deep learning solution to enable efficient quantification and analysis of wood cells suitable for practical applications.

cs.CV

Fabricating a dielectrophoretic microfluidic device using 3D-printed moulds and silver conductive paint

Dielectrophoresis is an electric field-based technique for moving neutral particles through a fluid. When used for particle separation, dielectrophoresis has many advantages compared to other methods, providing label-free operation with greater control of the separation forces. In this paper, we design, build, and test a low-voltage dielectrophoretic device using a 3D printing approach. This lab-on-a-chip device fits on a microscope glass slide and incorporates microfluidic channels for particle separation. First, we use multiphysics simulations to evaluate the separation efficiency of the prospective device and guide the design process. Second, we fabricate the device in PDMS (polydimethylsiloxane) by using 3D-printed moulds that contain patterns of the channels and electrodes. The imprint of the electrodes is then filled with silver conductive paint, making a 9 pole comb electrode. Lastly, we evaluate the separation efficiency of our device by introducing a mixture of 3 $\mu$m and 10 $\mu$m polystyrene particles and tracking their progression. Our device is able to efficiently separate these particles when the electrodes are energized with $\pm$12 V at 75 kHz. Overall, our method allows the fabrication of cheap and effective dielectrophoretic microfluidic devices using commercial off-the-shelf equipment.

physics.app-ph

Feedback Loops in Open Data Ecosystems

Public agencies are increasingly publishing open data to increase transparency and fuel data-driven innovation. For these organizations, maintaining sufficient data quality is key to continuous re-use but also heavily dependent on feedback loops being initiated between data publishers and users. This paper reports from a longitudinal engagement with Scandinavian transportation agencies, where such feedback loops have been successfully established. Based on these experiences, we propose four distinct types of data feedback loops in which both data publishers and re-users play critical roles.

cs.SE

Optical design for laser tweezers Raman spectroscopy setups for increased sensitivity and flexible spatial detection

We demonstrate a method to double the collection efficiency in Laser Tweezers Raman Spectroscopy (LTRS) by collecting both the forward and back-scattered light in a single-shot multitrack measurement. Our method can collect signals at different sample volumes, granting both the pinpoint spatial selectivity of confocal Raman and the bulk sensitivity of non-confocal Raman simultaneously. Further, we display that our approach allows for reduced detector integration time and laser power. Thus, our method will enable the monitoring of biological samples sensitive to high intensities for longer times. Additionally, we demonstrate that by a simple modification, we can add polarization sensitivity and retrieve extra biochemical information.

physics.optics

A Step-by-Step Guide to 3D Print Motorized Rotation Mounts for Optical Applications

Motorized rotation mounts and stages are versatile instruments that introduce computer control to optical systems, enabling automation and scanning actions. They can be used for intensity control and position adjustments, etc. However, these rotation mounts come with a hefty price tag, and this limits their use. This work shows how to build two different types of motorized rotation mounts for 1" optics, using a 3D printer and off-the-shelf components. The first is intended for reflective elements, like mirrors and gratings, and the second for transmissive elements, like polarizers and retarders. We evaluate and compare their performance to commercial systems based on velocity, resolution, accuracy, backlash, and axis wobble. Also, we investigate the angular stability using Allan variance analysis. The results show that our mounts perform similar to systems costing more than 2000 Euro, while also being quick to build and costing less than 200 Euro. As a proof of concept, we show how to control lasers used in an optical tweezers and Raman spectroscopy setup. When used for this, the 3D printed motorized rotational mounts provide intensity control with a resolution of $0.03$ percentage points or better.

physics.ins-det

Object Plane Detection and Phase Retrieval from Single-Shot Holograms using Multi-Wavelength In-Line Holograph

Phase retrieval and the twin-image problem in digital in-line holographic microscopy can be resolved by iterative reconstruction routines. However, recovering the phase properties of an object in a hologram needs an object plane to be chosen correctly for reconstruction. In this work, we present a novel multi-wavelength Gerchberg-Saxton algorithm to determine the object plane using single-shot holograms recorded with multiple wavelengths in an in-line holographic microscope. For micro-sized objects, we verify the object positioning capabilities of the method for various shapes and derive the phase information using synthetic and experimental data. Experimentally, we built a compact digital in-line holographic microscopy setup around a standard optical microscope with a regular RGB-CCD camera and acquire holograms of micro-spheres, E. coli and red blood cells, that are illuminated using three lasers operating at 491 nm, 532 nm and 633 nm, respectively. We demonstrate that our method provides accurate object plane detection and phase retrieval under noisy conditions, e.g., using low-contrast holograms without background normalization. This method allows for automatic positioning and phase retrieval suitable for holographic particle velocimetry, and object tracking in biophysical or colloidal research.

physics.optics

ToxTrac: a fast and robust software for tracking organisms

1. Behavioral analysis based on video recording is becoming increasingly popular within research fields such as; ecology, medicine, ecotoxicology, and toxicology. However, the programs available to analyze the data, which are; free of cost, user-friendly, versatile, robust, fast and provide reliable statistics for different organisms (invertebrates, vertebrates and mammals) are significantly limited. 2. We present an automated open-source executable software (ToxTrac) for image-based tracking that can simultaneously handle several organisms monitored in a laboratory environment. We compare the performance of ToxTrac with current accessible programs on the web. 3. The main advantages of ToxTrac are: i) no specific knowledge of the geometry of the tracked bodies is needed; ii) processing speed, ToxTrac can operate at a rate >25 frames per second in HD videos using modern desktop computers; iii) simultaneous tracking of multiple organisms in multiple arenas; iv) integrated distortion correction and camera calibration; v) robust against false positives; vi) preservation of individual identification if crossing occurs; vii) useful statistics and heat maps in real scale are exported in: image, text and excel formats. 4. ToxTrac can be used for high speed tracking of insects, fish, rodents or other species, and provides useful locomotor information. We suggest using ToxTrac for future studies of animal behavior independent of research area. Download ToxTrac here: https://toxtrac.sourceforge.io

cs.CV

UmUTracker: A versatile MATLAB program for automated particle tracking of 2D light microscopy or 3D digital holography data

We present a versatile and fast MATLAB program (UmUTracker) that automatically detects and tracks particles by analyzing video sequences acquired by either light microscopy or digital in-line holographic microscopy. Our program detects the 2D lateral positions of particles with an algorithm based on the isosceles triangle transform, and reconstructs their 3D axial positions by a fast implementation of the Rayleigh-Sommerfeld model using a radial intensity profile. To validate the accuracy and performance of our program, we first track the 2D position of polystyrene particles using bright field and digital holographic microscopy. Second, we determine the 3D particle position by analyzing synthetic and experimentally acquired holograms. Finally, to highlight the full program features, we profile the microfluidic flow in a 100 micrometer high flow chamber. This result agrees with computational fluid dynamic simulations. On a regular desktop computer UmUTracker can detect, analyze, and track multiple particles at 5 frames per second for a template size of 201 x 201 in a 1024 x 1024 image. To enhance usability and to make it easy to implement new functions we used object-oriented programming. UmUTracker is suitable for studies related to: particle dynamics, cell localization, colloids and microfluidic flow measurement.

cs.CV

A step-by-step guide to reduce spatial coherence of laser light using a rotating ground glass diffuser

Wide field-of-view imaging of fast processes in a microscope requires high light intensities motivating the use of lasers as light sources. However, due to their long spatial coherence length lasers are inappropriate for such applications as they produce coherent noise and parasitic reflections, such as speckle, degrading image quality. Therefore, we provide a step-by-step guide for constructing a speckle-free and high contrast laser illumination setup using a rotating ground glass diffuser driven by a stepper motor. The setup is easy to build, cheap and allows a significant light throughput of 48 %, which is 40 % higher in comparison to a single lens collector commonly used in reported setups. This is achieved by using only one objective to collect the scattered light from the ground glass diffuser. We validate the stability and performance of our setup in terms of image quality, motor-induced vibrations and light throughput. To highlight the latter, we record Brownian motion of micro-particles using a 100x oil immersion objective and a high-speed camera operating at 2 000 Hz with a laser output power of only 22 mW. Moreover, by reducing the objective magnification to 50x sampling rates up to 10 000 Hz are realized. To help readers with basic or advanced optics knowledge realizing this setup we provide; a full component list, 3D-printing CAD files, setup protocol, and the code for running the stepper motor.

physics.ins-det

Nonlinear Dynamic Force Spectroscopy

Dynamic force spectroscopy (DFS) is an experimental technique that is commonly used to assess information of the strength, energy landscape, and lifetime of noncovalent bio-molecular interactions. DFS traditionally requires an applied force that increases linearly with time so that the bio-complex under investigation is exposed to a constant loading rate. However, tethers or polymers can modulate the applied force in a nonlinear regime. For example, bacterial adhesion pili and polymers with worm-like chain properties are examples of structures that show nonlinear force responses. In these situations, the theory for traditional DFS cannot be readily applied. In this work we expand the theory for DFS to also include nonlinear external forces while still maintaining compatibility with the linear DFS theory. To validate the theory we modeled a bio-complex expressed on a stiff, an elastic and a worm-like chain polymer, using Monte Carlo methods, and assessed the corresponding rupture force spectra. It was found that the nonlinear DFS (NLDFS) theory correctly predicted the numerical results. We also present a protocol suggesting an experimental approach and analysis method of the data to estimate the bond length and the thermal off-rate.

physics.bio-ph

A robust particle detection algorithm based on symmetry

Particle tracking is common in many biophysical, ecological, and micro-fluidic applications. Reliable tracking information is heavily dependent on of the system under study and algorithms that correctly determines particle position between images. However, in a real environmental context with the presence of noise including particular or dissolved matter in water, and low and fluctuating light conditions, many algorithms fail to obtain reliable information. We propose a new algorithm, the Circular Symmetry algorithm (C-Sym), for detecting the position of a circular particle with high accuracy and precision in noisy conditions. The algorithm takes advantage of the spatial symmetry of the particle allowing for subpixel accuracy. We compare the proposed algorithm with four different methods using both synthetic and experimental datasets. The results show that C-Sym is the most accurate and precise algorithm when tracking micro-particles in all tested conditions and it has the potential for use in applications including tracking biota in their environment.

cs.CV

Circle detection using isosceles triangles sampling

Detection of circular objects in digital images is an important problem in several vision applications. Circle detection using randomized sampling has been developed in recent years to reduce the computational intensity. Randomized sampling, however, is sensitive to noise that can lead to reduced accuracy and false-positive candidates. This paper presents a new circle detection method based upon randomized isosceles triangles sampling to improve the robustness of randomized circle detection in noisy conditions. It is shown that the geometrical property of isosceles triangles provide a robust criterion to find relevant edge pixels and thereby efficiently provide an estimation of the circle center and radii. The estimated results given by the isosceles triangles sampling from each connected component of edge map were analyzed using a simple clustering approach for efficiency. To further improve on the accuracy we applied a two-step refinement process using chords and linear error compensation with gradient information of the edge pixels. Extensive experiments using both synthetic and real images were presented and results were compared to leading state-of-the-art algorithms and showed that the proposed algorithm: are efficient in finding circles with a low number of iterations; has high rejection rate of false-positive circle candidates; and has high robustness against noise, making it adaptive and useful in many vision applications.

cs.CV

Investigation of force approximations in tethered cells simulations

Simulations of tethered cells in viscous sub-layers are frequently performed using the Stokes drag force, but without taking into account contributions from surface corrections, lift forces, buoyancy, the Basset force, the cells finite inertia, or added mass. In this work, we investigate to which extent such contributions influence, under a variety of hydrodynamic conditions, the force at the anchor point of a tethered cell and the survival probability of a bacterium that is attached to a host by either a slip or a catch bond via a tether with a few different biomechanical properties. We show that a consequence of not including some of these contributions is that the force to which a bond is exposed can be significantly underestimated; in general by ~32-46 %, where the influence of the surface corrections dominate (the parallel and normal correction coefficients contribute with ~5-8 or 23-26 %, respectively). The Basset force is a major contributor, up to 20 %, for larger cells and shear rates. The lift force and inertia contribute when cells with radii >3 micrometer reside in shear rates >2000 1/s. Buoyancy contributes significantly for cells with radii >3 micrometer in shear rates <10 1/s. Since the lifetime of a bond depends strongly on the force, both the level of approximations and the biomechanical model of the tether significantly affect the survival probability of tethered bacteria. For a cell attached with a FimH-mannose bond and an extendable tether in a shear rate of 3000 1/s, neglecting the surface correction coefficients or the Basset force can imply that the survival probability is overestimated by more than an order of magnitude. This work thus shows that in order to quantitatively assess bacterial attachment forces and survival probabilities, both the fluid forces and the tether properties need to be modeled accurately.

physics.bio-ph

Cell shape identification using digital holographic microscopy

We present a cost-effective, simple and fast digital holographic microscopy method based upon Rayleigh-Sommerfeld back propagation for identification of the geometrical shape of a cell. The method was tested using synthetic hologram images generated by ray-tracing software and from experimental images of semi-transparent spherical beads and living red blood cells. Our results show that by only using the real part of the back-reconstructed amplitude the proposed method can provide information of the geometrical shape of the object and at the same time accurately determine the axial position of the object under study. The proposed method can be used in flow chamber assays for pathophysiological studies where fast morphological changes of cells are studied in high numbers and at different heights.

physics.optics

The biomechanical and structural properties of CS2 fimbriae

Enterotoxigenic Escherichia coli (ETEC) are a major cause of diarrhea worldwide, and infection of children in underdeveloped countries often leads to high mortality rates. Isolated ETEC express a plethora of colonization factors (fimbriae/pili), of which CFA/I and CFA/II that are assembled via the alternate chaperone pathway (ACP), are amongst the most common. Fimbriae are filamentous structures, whose shafts are primarily composed of helically arranged single pilin-protein subunits, with a unique biomechanical capability allowing them to unwind and rewind. A sustained ETEC infection, under adverse conditions of dynamic shear forces, is primarily attributed to this biomechanical feature of ETEC fimbriae. Recent understandings about the role of fimbriae as virulence factors are pointing to an evolutionary adaptation of their structural and biomechanical features. In this work, we investigated the biophysical properties of CS2 fimbriae from the CFA/II group. Homology modelling its major structural subunit CotA reveals structural clues and these are related to the niche in which they are expressed. Using optical tweezers force spectroscopy we found that CS2 fimbriae unwind at a constant force of 10 pN and have a corner velocity of 1300 nm/s, i.e., the velocity at which the force required for unwinding rises exponentially with increased speed. The biophysical properties of CS2 fimbriae assessed in this work classify them into a low-force unwinding group of fimbriae together with the CFA/I and CS20 fimbriae expressed by ETEC strains. The three fimbriae are expressed by ETEC, colonize in similar gut environments, and exhibit similar biophysical features, but differ in their biogenesis. Our observation suggests that the environment has a strong impact on the biophysical characteristics of fimbriae expressed by ETEC.

physics.bio-ph

Construction of force measuring optical tweezers instrumentation and investigations of biophysical properties of bacterial adhesion organelles

Optical tweezers are a technique in which microscopic-sized particles, including living cells and bacteria, can be non-intrusively trapped with high accuracy solely using focused light. The technique has therefore become a powerful tool in the field of biophysics. Optical tweezers thereby provide outstanding manipulation possibilities of cells as well as semi-transparent materials, both non-invasively and non-destructively, in biological systems. In addition, optical tweezers can measure minute forces (< 10-12 N), probe molecular interactions and their energy landscapes, and apply both static and dynamic forces in biological systems in a controlled manner. The assessment of intermolecular forces with force measuring optical tweezers, and thereby the biomechanical structure of biological objects, has therefore considerably facilitated our understanding of interactions and structures of biological systems. Adhesive bacterial organelles, so called pili, mediate adhesion to host cells and are therefore crucial for the initial bacterial-cell contact. Thus, they serve as an important virulence factor. The investigation of pili, both their biogenesis and their expected in vivo properties, brings information that can be of importance for the design of new drugs to prevent bacterial infections, which is crucial in the era of increased bacterial resistance towards antibiotics. In this thesis, an experimental setup of a force measuring optical tweezers system and the results of a number of biomechanical investigations of adhesive bacterial organelles are presented. Force measuring optical tweezers have been used to characterize three different types of adhesive organelles under various conditions, P, type 1, and S pili, which all are expressed by uropathogenic Escherichia coli. A quantitative biophysical force-extension model, built upon the structure and force response, has been developed.

physics.bio-ph

The biomechanical properties of F1C pili

Uropathogenic Escherichia coli (UPEC) express various kinds of organelles, so-called pili or fimbriae, that mediate adhesion to host tissue in the urinary tract through specific receptor-adhesin interactions. The biomechanical properties of these pili have been considered important for the ability of bacteria to withstand shear forces from rinsing urine flows. Force measuring optical tweezers have been used to characterize individual organelles of F1C type expressed by UPEC bacteria with respect to such properties. Qualitatively, the force-vs.-elongation response was found to be similar to that of other types of helix-like pili expressed by UPEC, i.e. type 1, P, and S, with force-induced elongation in three regions of which one represents the important uncoiling mechanism of the helix-like quaternary structure. Quantitatively, the steady-state uncoiling force was assessed to 26.4(1.4) pN, which is similar to those of other pili (which range from 21 pN for SI to 30 pN for type 1). The corner velocity for dynamic response (1400 nm/s) was found to be larger than those of the other pili (400 -700 nm/s for S and P pili, and 6 nm/s for type 1). The kinetics were found to be faster, with a thermal opening rate of 17 Hz, a few times higher than S and P pili, and three orders of magnitude times higher than type 1. These data suggest that F1C pili are, like P and S pili, evolutionary-selected to primarily withstand the conditions expressed in the upper urinary tract.

physics.bio-ph

Damping properties of type 1 fimbriae

Type 1 fimbriae mediate adhesion of uropathogenic Escherichia coli (UPEC) to host cells. It has been hypothesized that fimbriae can, by their ability to uncoil under exposure to force, reduce fluid shear stress on the adhesin-receptor interaction by which the bacterium adheres to the surface. In this work we develop a model that describes how the force on the adhesin-receptor interaction of a type 1 fimbriae varies as a bacterium is affected by a time dependent fluid flow mimicking in vivo conditions. The model combines in vivo hydrodynamic conditions with previously assessed biomechanical properties of the fimbriae. Numerical methods are used to solve for the motion and adhesion force under the presence of time dependent fluid profiles. It is found that a bacterium tethered with a type 1 pilus will experience significantly reduced shear stress for moderate to high flow velocities and that the maximum stress the adhesin will experience is limited to ~120 pN, which is sufficient to activate the conformational change of the FimH adhesin into its stronger state but also lower than the force required for breaking it under rapid loading. Our model thus supports the assumption that the type 1 fimbriae shaft and the FimH adhesin-receptor interaction are optimized to each other, and that they give piliated bacteria significant advantages in rapidly changing fluidic environments.

physics.bio-ph