![]() Indeed, in the field of theoretical physics there has been an evolution of the concept of reality: from Galileo’s famous sentence “(the universe) it is written with mathematical language” to the most modern interpretations, according to which “the universe we live in is itself a mathematical structure” and more importantly is computable. 4.2c shows saturation (peak at the maximum value of 255).Ĥ.2.1 What Is the Difference Between a Material Object and a Representation of It?ĭuring imaging and analysis of the resulting image-type datasets, it should never be forgotten that images are a “representation” of reality. 4.2a shows an overall low signal and in f) the histogram of the image Fig. The real intensity distributions can be comprehended by using the histogram, where the pixel values are plotted: in Fig. Increasing the contrast might render similar to the eye two images acquired with hugely different settings (in this case Fig. 4.2 the same image is visualized with (a) low and (d) high contrast. The image can be visualized with settings that might induce the observer to draw wrong conclusions or can be easily altered by unattentive manipulation, or even worse, by mischievous practices. Figure 4.2 shows how images of the same mouse tissue can differ if acquired with (a) low, (b) intermediate, and (c) high gain.ĭata handling, processing, and visualization settings. A few examples of conditions that can change during image acquisition in microscopy are illumination level and evenness, detectors gain, optics alignment, image format (bit depth, type), etc. The image itself is determined by sample preparation protocols and acquisition conditions that not always can be set in a reproducible way from session to session. This is not only to eliminate the subjective bias of the observer but also to take into account the possible variability between different imaging sessions:Įxperimental conditions and image acquisition parameters. Light microscopy allows wonderful discoveries about the shape and functions of biological samples, but beyond the descriptive power, it is necessary to apply reproducible acquisition settings and quantitative methods to analyze the resulting images. In biology, the use of fluorescence resonant energy transfer can be used to obtain the stoichiometry of interacting molecules in living cells, where typical dimensions are in the order of nanometers.Ĥ.1.2 Need for Quantitative Methods Is Extended to Light Microscopy In astronomy, the radiation from the cosmos is detected to measure which wavelengths of the light spectrum are missing (absorption bands) it is then possible to determine the atomic composition of stars which are billions of kilometers distant from the Earth. Therefore, the image, considered as the intensity of detected radiation, is not simply a qualitative description of an object or a biological sample, but has the potentiality to convey useful and unexpected information, often about very different scientific aspects, and so it becomes a “measurable entity.” Footnote 2Įxamples of scientific applications of imaging can come from extremely different disciplines: 4.1 shows BPAE cells properly stained to characterize different cellular compartments: nucleus, cytoplasm, and mitochondria. Modern science can, fortunately, rely on detectors to capture images of the reality (thanks mainly to the theoretical formulation and experiments of those “analogical” geniuses such as Galilei, Newton, and Einstein Footnote 1).Ī variety of imaging techniques, some more invasive than others, can be used to study the human anatomy or functional aspects of cellular organelles at a different scale of resolution. From the first attempts made by Renaissance artists to accurately describe the human body anatomy to the representation of the lunar topography done by Galileo Galilei, the common surprising feature is the effort to render the image as representative as possible of the investigated object. The use of the image as a vector of information is not a feature that uniquely belongs to modern science. 4.1.1 Image Relevance in Science from Biology to Astronomy ![]()
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