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Schematic of the high-resolution-UC2 widefield microscope. a. Overview of the different component categories of the microscope, from the hardware, through the electronics controlling, to the software and the ImSwitch-based GUI. b. Schematic of the complete setup. Red and blue stars are references to help visualize the 3D structure. Top layer is the commercially bought XY-stage. Timing belts convert motor torque (two grey gears top left) into stage motion (bigger black gear middle) but have been omitted in this representation for clarity. Sample holder can be printed according to the sample used, circular in this case. c. Commercially bought precision Z-stage and high-NA objective (Olympus 60x/1.49 NA TIRF) d. Detection layer, corresponds to the bottom layer in the setup (red star as reference). Emitted fluorescence (dark red light-path) is depicted with a f = 100 mm lens on the detector (Alvium 1800 U-158c CMOS camera from Allied
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Fluorescence microscopy is a fundamental tool in the life sciences, but the availability of sophisticated equipment required to yield high-quality, quantitative data is a major bottleneck in data production in many laboratories worldwide. This problem has long been recognized and the abundancy of low-cost electronics and the simplification of fabri...
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... an intuitive and user-friendly GUI for maximal accessibility. Lastly, the entire system including the physical microscope, the electronics and the software must be open-source and be possible to assemble and operate with moderate technical know-how (Figure 1a ). We reasoned that the quality of the objective is paramount to image quality, while most other optical parts of a light microscope can be mass-produced components. ...
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... on the guidelines described above and to ensure adaptability and modularity, we decided to build our open-source automated microscope on the basis of the UC2 toolbox using its injection molded cubes for the structural assembly. For ease of usage, we used an inverted configuration and assembled the microscope in three layers to keep the size and design compact (Figure 1b ). The high-precision x-y sample table and z-stage ( Figure 1c ) were low-cost commercial solutions that we motorized and implemented into the open-source, python-based microscope GUI as automated elements of the microscope. ...
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... ease of usage, we used an inverted configuration and assembled the microscope in three layers to keep the size and design compact (Figure 1b ). The high-precision x-y sample table and z-stage ( Figure 1c ) were low-cost commercial solutions that we motorized and implemented into the open-source, python-based microscope GUI as automated elements of the microscope. The emission pathway at the bottom of the microscope assembly is based on an Alvium 1800 U-158c CMOS camera from Allied Vision, which compared to conventional scientific cameras is more affordable by a factor of 30 to 60 times at the time of submission (Figure 1d ), more compact and consumes less energy. ...
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... high-precision x-y sample table and z-stage ( Figure 1c ) were low-cost commercial solutions that we motorized and implemented into the open-source, python-based microscope GUI as automated elements of the microscope. The emission pathway at the bottom of the microscope assembly is based on an Alvium 1800 U-158c CMOS camera from Allied Vision, which compared to conventional scientific cameras is more affordable by a factor of 30 to 60 times at the time of submission (Figure 1d ), more compact and consumes less energy. One important point here is that the camera needs to be monochromatic as the Bayer pattern on a polychromatic camera would significantly reduce the sampling as well as the quantum efficiency of a camera chip, which is already lower compared to that of scientific cameras. ...
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... camera was connected to the computer via USB 3.0, allowing fast data transfer and maximal compatibility. Between the emission pathway and the sample stage, the excitation pathway contained a low-cost 635 nm diode laser with an optional beam expander (Figure 1e ), that depending on the illumination requirements could be inserted or removed, modulating the illumination density and homogeneity of the excitation field (see Figure 2c ). To trigger the laser, control laser power and to steer the x, y and z motors for focus and sample translation, we used a low-cost Arduino-coded board that can be connected via USB3.0 to any computer. ...
Citations
Microscopy is essential for detecting, identifying, analyzing, and measuring small objects. Access to modern microscopy equipment is crucial for scientific research, especially in the biomedical and analytical sciences. However, the high cost of equipment, limited availability of parts, and challenges associated with transporting equipment often limit the accessibility and operational capabilities of these tools, particularly in field sites and other remote or resource-limited settings. Thus, there is a need for affordable and accessible alternatives to traditional microscopy systems. We address this challenge by investigating the feasibility of using a simple microcontroller board not only as a portable and field-ready digital microscope, but furthermore as a versatile platform which can easily be adapted to a variety of imaging applications. By adding a few external components, we demonstrate that a low-cost ESP32 camera board can be used to build an autonomous in situ platform for digital time-lapse imaging of cells. Our prototype of this approach, which we call ESPressoscope, can be adapted to applications ranging from monitoring incubator cell cultures in the lab to observing ecological phenomena in the sea, and it can be adapted for other techniques such as microfluidics or spectrophotometry. Our prototype of the ESPressoscope concept achieves a low power consumption and small size, which makes it ideal for field research in environments and applications where microscopy was previously infeasible. Its Wi-Fi connectivity enables integration with external image processing and storage systems, including on cloud platforms when internet access is available. Finally, we present several web browser-based tools to help users operate and manage our prototype’s software. Our findings demonstrate the potential for low-cost, portable microscopy solutions to enable new and more accessible experiments for biological and analytical applications.
Developing devices and instrumentation in a bioimaging core facility is an important part of the innovation mandate inherent in the core facility model but is a complex area due to the required skills and investments, and the impossibility of a universally applicable model. Here, we seek to define technological innovation in microscopy and situate it within the wider core facility innovation portfolio, highlighting how strategic development can accelerate access to innovative imaging modalities and increase service range, and thus maintain the cutting edge needed for sustainability. We consider technology development from the perspective of core facility staff and their stakeholders as well as their research environment and aim to present a practical guide to the ‘Why, When, and How’ of developing and integrating innovative technology in the core facility portfolio.
Core facilities need to innovate to stay up to date. However, how to carry out the innovation is not very obvious. One area of innovation in imaging core facilities is the building of optical setups. However, the creation of optical setups requires specific skill sets, time, and investments. Consequently, the topic of whether a core facility should develop optical devices is discussed as controversial. Here, we provide resources that should help get into this topic, and we discuss different options when and how it makes sense to build optical devices in core facilities. We discuss various aspects, including consequences for staff and the relation of the core to the institute, and also broaden the scope toward other areas of innovation.
The field of microscopy has been empowering humankind for many centuries by enabling the observation of objects that are otherwise too small to detect for the naked human eye. Microscopy techniques can be loosely divided into three main branches, namely photon-based optical microscopy, electron microscopy, and scanning probe microscopy with optical microscopy being the most prominent one. On the high-end level, optical microscopy nowadays enables nanometer resolution covering many scientific disciplines ranging from material sciences over the natural sciences and life sciences to the food sciences. On the lower-end level, simplified hardware and openly available description and blueprints have helped to make powerful microscopes widely available to interested scientists and researchers. For this special issue, we invited contributions from the community to share their latest ideas, designs, and research results on open-source hardware in microscopy. With this collection of articles, we hope to inspire the community to further increase the accessibility, interoperability, and reproducibility of microscopy. We further touch on the standardization of methodologies and devices including the use of computerized control of data acquisition and data analysis to achieve high quality and efficiency in research and development.
