Synopsis
The microscope’s resolution greatly influences the value and effectiveness of the pathological process. Therefore, it is important for people practicing digital pathology to handle it very carefully. While the concept of microscope resolution seems simple at first glance, it turns out to be quite complex and has many facets. The complexity comes from the fact that various things affect resolution, which means that individuals need to know these hurdles comprehensively. To understand this depth and importance of microscope resolution in digital pathology, it is necessary to gather more information on these topics. As a pathologist, you work with specimens of diverse sizes and every one of them requires a different resolution. If you apply the wrong one, you can miss all the details that need to be observed. This article will delve into the concepts and factors of microscope resolution.
Microscope Resolution in Digital Pathology
In digital pathology, you need to pay attention to all the concepts, factors, and calculations that concern the microscope’s resolution. When selecting the resolution, you need to determine the level of detail that needs to be seen as some parts of cells and bacteria are very tiny. Interpreting the cells and bacteria is critical in researching or setting up diagnoses. By increasing the resolution, you can identify smaller substructures and study the case at different angles. With increased resolution, you can distinguish between the different types of cells, identify structural abnormalities, and observe changes that happen during biological processes such as mitosis or cell division. Biology, medicine, materials science, and others are disciplines where the right microscopy resolution can be of great help to study all the important phenomena.
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Why is Microscope Resolution Important?
Resolution can be defined as the measure of the smallest distance between two objects that can be distinguished as separate objects. Simply said, the higher the resolution, the more details can be seen in the image. In the field of digital pathology, microscope resolution is of special importance because experts often encounter structures of very small and varying sizes.
Factors Influencing Resolution
The wavelength of the light with which the object is illuminated is one of the most important factors. It determines the smallest distance that can be resolved with the microscope. When light passes through a small aperture such as the aperture of a microscope, it is naturally scattered or diffracted. The resolution of the microscope can be reduced by diffraction due to the scattering of the light. Another important point is the lenses or mirrors. To avoid distortion, they should be manufactured more precisely. This can lead to blurred images and poor quality if the light is a little out of focus or other imaging errors are present. The quality of the performance is affected by possible blurring or distortion. These issues can occur in the event of misbehavior of some optical elements such as lenses, where the numerical aperture (NA) measures the light gathering capacity. You can calculate the resolution of a microscope by using the Abbe equation:
resolution = 0.61 x (wavelength / NA)
The resolution is specified in distance units such as nanometers, the wavelength is the light’s wavelength, and NA is the lens’ numerical aperture.
The equation shows the relationship of the resolution of the microscope, the wavelength of the light used, NA, and the refractive index.
Microscope Resolution in Science
Digital pathology is about microscopic things, as we move into the field of nanoscience, including the nanotechnology base. These are just some of the areas that are increasingly going hand in hand over time. The same applies to the two areas of industry and science. The search for and identification of atomic configurations in nanostructures has become old-fashioned with this generation. The electron microscope has an intrinsic capacity for atomic scale imaging as well as for chemical analysis and hence it has become a tool that is indispensable to material pathologists.
Challenges in Using Microscope Resolution
Microscope resolution, while a cornerstone of scientific investigation, presents a fascinating array of challenges that researchers must contend with. Overcoming these hurdles not only demands innovative instrumentation but also necessitates a deep understanding of the intricacies of light diffraction, aberrations, and other factors.
Diversity of Terms
Regarding resolution, there is still a certain source of confusion among microscopists, and it mostly involves a plethora of terms used to define resolution itself. Some experts connect it with the phase-contrast transfer function. The instrumental resolution of the microscope is often termed the information limit, and it can extend to much higher values of spatial frequency than the structural resolution when the electron beam incident onto the sample is highly coherent, although phase inversions caused by TH oscillations will then make it difficult to interpret very fine image features reliably. Lattice fringe resolution describes the interference between two or more diffracted beams from a crystalline material. This resolution limit mainly reflects the stability of the microscope and the quality of its environment. It is important to pay attention to the difference between lattice-fringe imaging, and atomic column imaging. Precise information about atomic column arrangements can only be determined for a small focal range and quite limited crystal thickness.
Aberration Correction
Aberration correction is one of the greatest challenges medical experts and microscopists face. Functional hardware correctors such as double-hexapole correctors and multiple quadrupole-octupole systems were of great help in overcoming these challenges. They contributed significantly to the overall success of the development of resolution enhancement because considerable image detail present beyond the structural resolution limit became directly interpretable once aberration correction had been accomplished. In digital holographic microscopy, resolution enhancement is accomplished by illumination modulation techniques, such as oblique illumination, structured illumination, and speckle illumination. All of this can enhance the resolution by adding more high-frequency information to the recording system. Resolution enhancement can also be achieved by extrapolation of a hologram or by synthesizing a larger hologram by scanning the sample, the camera, or inserting a diffraction grating between the sample and the camera. For on-chip digital holographic microscopy, spatial resolution is usually achieved using pixel super-resolution techniques.
Conclusion
The importance of microscope resolution for every digital pathology workflow is gigantic. The concept of microscope resolution can be explained simply: the greater the resolution, the more details on the specimen can be seen more easily. In the field of digital pathology details are everything. But, as those details are of different sizes, you need to be familiar with how to determine the best resolution. Abbe equation helps achieve the right resolution to be used. Microscope resolution is more and more mentioned in the context of nanoscience or digital holographic microscopy. Lattice fringe resolution reflects the stability of the microscope, while in digital holographic microscopy illumination modulation techniques enhance the resolution. In setting up the proper microscope resolution for your workflow, digital microscopes can be of great help.
