Controlled liquid exchange is a fundamental requirement for numerous biological and chemical experimental protocols. However, achieving constant-volume exchange often requires electronic syringe pumps that are cost-prohibitive and time-consuming to set up in a push-pull configuration. To address these limitations, we developed the Push2Pull syringe holder, a simple 3D-printed device that mechanically synchronizes two syringes to simultaneously add and extract equal fluid volumes. This device is compatible with standard disposable syringes from 1 to 60 mL in size and operates without electricity or additional hardware, making it ideal for both laboratory and field settings. Validation experiments demonstrate an exchange accuracy within ±2%v/v across the whole travel range, while fluid exchange efficiency was calculated for various use cases using CFD simulations. The Push2Pull syringe holder offers an accessible, open-source solution for precise fluid handling, for a material cost of less than $10.
Growing biofilms of thermophilic (heat-loving) and psychrotrophic (cold-tolerant) bacteria pose several challenges due to specific environmental requirements. Thermophilic bacteria typically grow between 45 and 80 C, while psychrotrophic bacteria thrive between 0 and 15 C. Maintaining the precise temperature and fluid conditions required for biofilm growth can be technically challenging. To overcome these challenges, we designed the Bio-Rocker, a temperature- and shear stress-controlled rocker platform for biofilm incubation. The platform supports temperatures between − 9 and 99 C, while its digital controller can adjust the rocking speed from 1 to 99/s and set rocking angles up to ±19. This ability, together with data from analytical models and multi-physics simulations, provides control over the shear stress distribution at the growth surfaces, peaking at 2.4 N/m. Finally, we evaluated the system’s ability to grow bacteria at different temperatures, shear stress, and materials by looking at the coverage and thickness of the biofilm, as well as the total biomass. A step-by-step guide, 3D CAD files, and controller software is provided for easy replication of the Bio-Rocker, using mostly 3D-printed and off-the-shelf components. We conclude that the Bio-Rocker’s performance is comparable to high-end commercial systems like the Enviro-Genie (Scientific Industries) yet costs less than $350 dollars to produce.
Analyzing microscopy images of large growing cell samples using traditional methods is a complex and time-consuming process. In this work, we have developed an attention-driven UNet-enhanced model using deep learning techniques to efficiently quantify the position, area, and circularity of bacterial spores and vegetative cells from images containing more than 10,000 bacterial cells. Our attention-driven UNet algorithm has an accuracy of 96%, precision of 82%, sensitivity of 81%, and specificity of 98%. Therefore, it can segment cells at a level comparable to manual annotation. We demonstrate the efficacy of this model by applying it to a live-dead decontamination assay. The model is provided in three formats: Python code, a Binder that operates within a web browser without needing installation, and a Flask Web application for local use.
Bacterial spores are problematic in agriculture, the food industry, and healthcare, with the fallout costs from spore-related contamination being very high. Spores are difficult to detect since they are resistant to many of the bacterial disruption techniques used to bring out the biomarkers necessary for detection. Because of this, effective and practical spore disruption methods are desirable. In this study, we demonstrate the efficiency of a compact microfluidic lab-on-chip built around a coplanar waveguide (CPW) operating at 2.45 GHz. We show that the CPW generates an electric field hotspot of ∼10 kV/m, comparable to that of a commercial microwave oven, while using only 1.2 W of input power and thus resulting in negligible sample heating. Spores passing through the microfluidic channel are disrupted by the electric field and release calcium dipicolic acid (CaDPA), a biomarker molecule present alongside DNA in the spore core. We show that it is possible to detect this disruption in a bulk spore suspension using fluorescence spectroscopy. We then use laser tweezers Raman spectroscopy (LTRS) to show the loss of CaDPA on an individual spore level and that the loss increases with irradiation power. Only 22% of the spores contain CaDPA after exposure to 1.2 W input power, compared to 71% of the untreated control spores. Additionally, spores exposed to microwaves appear visibly disrupted when imaged using scanning electron microscopy (SEM). Overall, this study shows the advantages of using a CPW for disrupting spores for biomarker release and detection.
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, like 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 μm and 10 μm polystyrene particles and tracking their progression. Our device is able to efficiently separate these particles when the electrodes are energized with ±12 V at 75 kHz. Overall, our method allows the fabrication of cheap and effective dielectrophoretic microfluidic devices using commercial off-the-shelf equipment.
Spore-forming pathogenic bacteria are adapted for adhering to surfaces, and their endospores can tolerate strong chemicals making decontamination difficult. Understanding the physico-chemical properties of bacteria and spores is therefore essential in developing antiadhesive surfaces and disinfection techniques. However, measuring physico-chemical properties in bulk does not show the heterogeneity between cells. Characterizing bacteria on a single-cell level can thereby provide mechanistic clues usually hidden in bulk measurements. This paper shows how optical tweezers can be applied to characterize single bacteria and spores, and how physico-chemical properties related to adhesion, fluid dynamics, biochemistry, and metabolic activity can be assessed.
Visualizing medical images from patients as physical 3D models (phantom models) have many roles in the medical field, from education to preclinical preparation and clinical research. However, current phantom models are generally generic, expensive, and time-consuming to fabricate. Thus, there is a need for a cost- and time-efficient pipeline from medical imaging to patient-specific phantom models. In this work, we present a method for creating complex 3D sacrificial molds using an off-the-shelf water-soluble resin and a low-cost desktop 3D printer. This enables us to recreate parts of the cerebral arterial tree as a full-scale phantom model (10×6×410×6×4 cm) in transparent silicone rubber (polydimethylsiloxane, PDMS) from computed tomography angiography images (CTA). We analyzed the model with magnetic resonance imaging (MRI) and compared it with the patient data. The results show good agreement and smooth surfaces for the arteries. We also evaluate our method by looking at its capability to reproduce 1 mm channels and sharp corners. We found that round shapes are well reproduced, whereas sharp features show some divergence. Our method can fabricate a patient-specific phantom model with less than 2 h of total labor time and at a low fabrication cost.
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, precision, 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.