Microscopy and micromanipulation technologies that include robotics have immensely contributed to the progress of cell manipulation techniques. Robotics-assisted microscopy combines high-content screening methods that enable the application of multivariate experimental approaches associated with large cell populations with independent cell-level sensitivity. Specific microscopes, such as the Nikon Ti2 line, are suitable for robotics-assisted microscopy applications.
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Before the robotic microscope development, the immunocytochemistry technique was used to view neurons. However, this technique had many limitations, including the ability to generate only a snapshot of cells at a single point in time without providing any information about the neurodegenerative process. Also, traditional methods of cellular analysis are laborious and time-consuming.
Robotization of the procedures in microscopy and cell manipulation entails the development of highly intelligent images that have immensely helped life science research. Two of the parameters that must be considered for this type of microscopy are camera calibration and image-motion coordinates in accordance with the microscope images.
General Working and Development of Robotics Microscope
Robotics-assisted microscopy, which is also referred to as robotic microscopy, is considered to be the brainchild of Steven Finkbeiner, an investigator at the Gladstone Institute of Neurological Disease. Dr. Finkbeiner, who is a pioneer in the field of Robotic Microscopy, developed this microscope to facilitate his research on Huntington's disease.
Although a robotic microscope looks like an ordinary inverted microscope, there are many dissimilarities in relation to their speed, motors, and computer programs that can automatically focus the object, move the stage, and can provide images of the cells growing in a tissue culture plate.
The microscope first focuses on the internal reference point in a plate of cells. Following this, it moves the plate a precise distance and refocuses automatically, and takes photographs. It repeats this process until it captures images of the entire plate or target area. Importantly, when the same plate is re-exposed to the same robotic microscope, even many days later, it can identify the same cells and re-examine them.
A computer program Dr. Finkbeiner developed could automatically analyze the obtained images rapidly, i.e., within minutes. This program can measure cells with specific morphologies, distinct protein content, or other features.
The robotic microscope has enabled the analysis of neuronal cells that express mutant huntingtin labeled with fluorescent reporter molecules. The fluorescence emitted from the mutant protein can be easily visualized via special filters on the microscope. Through repeated examination of the same cells, Dr. Finkbeiner was able to link the appearance and aggregation of the mutant huntingtin protein with cellular changes indicative of degeneration.
Main Functions of Robotics-Assisted Microscopy
Robotics-assisted microscopy combines multiple technologies for automated imaging and high throughput analysis. Some of its primary functions include multi-parametric and multi-channel detection of targeted cells or molecules and automated sample translation for serial imaging. Robotic microscopy has also played an important role in microinjection, where the insertion point must be precise with the imaging plane.
Robotics-assisted microscopy entails micromanipulations based on the user command alongside robotic vision and manipulation assistance. Three specific modules are implemented in this type of microscope, namely, a user-specified target tracker, vision-guided manipulation, and similarity-score-based adaptive compensation.
The user-specified target tracker helps to select a specific target to be tracked from the microscope display. Typically, this target is at the tip of a micropipette needle or holder, connected to the micromanipulator. After manipulating the desired target, the system restores the template, which is analyzed via high throughput screening and other procedures.
Robotic microscopy is linked to the removal and replacement of multiple culture vessels. It also allows the analysis of individual live cells amongst high-density cell populations. Compared to a pre-configured system, robotic microscopes are configured on conventional microscopes. It is more advantageous due to its flexible platform. The flexibility allows the users to exchange components, such as optics, emission splitters, detectors, and filter wheels, as per their needs.
Utilization of Robotic Microscope-Nikon TE-2000 to Access Treatment of Amyotrophic Lateral Sclerosis (ALS) Disease
Robotic microscopy has been used to assess the treatment of ALS, which is a motor neuron disease where the neurons degenerate and die. This disease is caused by mutations to TARDBP, also known as TDP43. Studies have shown that increased expression of both wild-type and mutant TDP43 correlates with decreased cell survival.
Scientists have used robotic microscopy to screen a large drug library on ALS models. This study used a Nikon TE-2000 microscope with Perfect Focus, essential for rapid, accurate focusing and reduction in acquisition times and phototoxicity.
Notably, the Nikon Ti2-E features the new Perfect Focus System 4 with a 25-mm-FOV camera port that enables faster and more accurate focusing than before. This advancement is extremely important for multipoint imaging in high-content applications. Since the Ti2's larger FOV enables a reduction in the number of images required to achieve improved results, it is an incredibly scalable and cost-effective tool in life science research.
Digitalizing Robotic Microscope
Scientists at Oldenburg University have digitalized robotic microscope for life science research. The main feature of this microscope is that it can record, scan and document fine tissue samples automatically, much faster than humans. Prof. Dr. Henrik Mouritsen, a neurobiologist who used this microscope stated, “With this machine what used to take a month can be done in a single night, and in a better quality than we ever had before”.
Interestingly, this microscope was used to study how migratory birds utilize their magnetic sense and other sensory inputs for navigation. Researchers at the Sensory Biology of Animals use this instrument regularly for different experiments, such as to analyze the magnetic sense of salmon. In addition, the development of deafness and the functioning of auditory pathways in mice have been studied using a robotic microscope.
References and Further Reading
New Robotic Microscope Helps Scientists Track Cells Over Time. (2022) ScienceDaily. [Online] Available at: https://www.sciencedaily.com/releases/2002/06/020610074158.htm
Joel, T. et al. (2020) Robotic microscopy for everyone: the OpenFlexure microscope. Biomedical Optics Express, 11, pp. 2447-2460. https://doi.org/10.1364%2FBOE.385729
A New Type of Robotic Microscope. (2017) [Online] Available at: https://www.medicaldesignandoutsourcing.com/a-new-type-of-robotic-microscope/
L. Yang, et al. (2016) Towards automatic robot-assisted microscopy: An uncalibrated approach for robotic vision-guided micromanipulation, 2016 IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS), Daejeon, Korea (South), pp. 5527-5532, https://doi.org/10.1109/IROS.2016.7759813.
Parekattil, S.J. and Moran, M.E. (2010) Robotic instrumentation: Evolution and microsurgical applications. Indian Journal of Urology. 26(3), pp. 395-403. https://doi.org/10.4103%2F0970-1591.70580.