MICRO-STRUCTURE ANALYSIS OF PLANT TISSUES - Lublin

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In 1830, Lister published the theoretical foundations for building achromatic lenses. His ideas were then employed by other microscope designers [3,14]. Moreover, Giovanni B. Amici discovered a method for the elimination of spherical aberration through the application of a suitable combination of lenses. The two discoveries permitted the building of multi-lens objective lenses and condensers which gave greater magnification ratios while retaining high optical correctness level. In 1827, Amici invented the immersion lens [12,14]. Also in 1872 the German physicist, Ernst Abbe, equipped the microscope with a lamp [13]. At the beginning pf the 20 th century the optical microscope was capable of magnification ratios of about x2000. In 1931, a team of German physicists headed by Ernst Rusk (who was awarded the Nobel Prize for that work, in 1986), designed the electron microscope, a usable version of which was built in 1938 by the Siemens company. The microscope permits magnification ratios of the order of x250,000. In 1942, C. Magnan (France) invented so-called proton microscope, the theoretical capabilities of which reach to the level of x5000,000 magnification ratio; in practice, however, its performance is no better than that of the electron microscope. In 1956, the American Ervin W. Mueller designed an ion microscope for examination of the structure of metallic blades, permitting magnification ratios of the order of several million. In 1962, E.N. Leith and J. Upatnieks designed and built a lens-less holographic optical microscope [3, 6, 12, 13, 14]. In spite of the tremendous progress in the field of microscopy, work is continued on better still and more modern microscope designs, yielding better and better effects of observation. 2.1.1. Principle of optical microscope The structure of the optical microscope utilizes two systems – the optical system, used for illumination of the object observed and for image magnification, and the mechanical system, the function of which is to ensure correct positioning of the particular components of the optical system. The key aspect is stability, mutual parallelism and concentricity of the optical system components. Fig. 2.4 presents the structure of the optical microscope. 16
3 5 9 6 7 8 4 2 1 Fig. 2.4. The structure of the optical microscope: 1 – condenser, 2 – objective lens, 3 – eyepiece, 4 – mirror or lamp, 5 – CCD camera, 6 – moving or mechanical stage with knobs for slide position control in the XY plane, 7 – coarse adjustment knob, 8 – fine adjustment knob, 9-revolving nosepiece (with objective lenses). 2.1.2. Optical elements Illumination system: in ordinary microscopes it will be a mirror; it can also be a simple lamp with a reflecting surface, or a sophisticated illumination system with a collector, position adjustment, individual power supply with voltage regulation, etc. Condenser: the condenser lenses concentrate the light coming from the mirror on the object (specimen) under observation. The image generated in the microscope is magnified and reversed with relation to the actual object. The microscope magnifies also the angle of view of the object under observation. Immersion: filling the space between the slide and cover and the objective and condenser with a liquid. Objective lenses: systems of lenses receiving the light from the slide and generating its magnified image. Body tube: - the tube in which the objective and the eyepiece are installed and where the image is generated. 17
Page 1 and 2: MICRO-STRUCTURE ANALYSIS OF PLANT T
Page 3 and 4: MICRO-STRUCTURE ANALYSIS OF PLANT T
Page 5 and 6: PREFACE The subject matter of resea
Page 7 and 8: PREFACE ...........................
Page 9 and 10: 1. THE GOAL OF IMAGE ANALYSIS OF PL
Page 11 and 12: within an object, caused e.g. by a
Page 13 and 14: 1.4. Analysis of plant tissue struc
Page 15 and 16: 2. MICROSCOPES AND SAMPLE PREPARATI
Page 17: focusing mechanism permitting the a
Page 21 and 22: 2.1.3. Mechanical elements Microsco
Page 23 and 24: 2.1.4. Image generation in optical
Page 25 and 26: 2.1.5. Resolution of the microscope
Page 27 and 28: a) 1 b) Relative refractive index 3
Page 29 and 30: 2.1.7. Microscopy studies Microscop
Page 31 and 32: One should take care, however, that
Page 33 and 34: may be difficult and the objects fi
Page 35 and 36: Object slicing and application of s
Page 37 and 38: them to straighten out without dama
Page 39 and 40: Fig. 2.15. Reconstruction of the tr
Page 41 and 42: Fig. 2.16. Images of the same part
Page 43 and 44: Fig. 2.17. Images of the structure
Page 45 and 46: 2.2. Confocal Microscopy Artur Zdun
Page 47 and 48: Fig. 2.19. Potato tuber tissue. The
Page 49 and 50: Fig. 2.21. Confocal Microscope oper
Page 51 and 52: Fig. 2.22. Potato tuber tissue. Ima
Page 53 and 54: in spite of the different original
Page 55 and 56: estimated that for most animal tiss
Page 57 and 58: However, because of the broadening
Page 59 and 60: 2.3. Electron Microscopy Andrzej Kr
Page 61 and 62: Due to the high vacuum inside the m
Page 63 and 64: Fig. 2.38 presents an image of aggr
Page 65 and 66: Fig. 2.38. Aggregation of starch gr
Page 67 and 68: a) b) c) d) Fig. 2.40. Schematics o
Page 69 and 70:
Fig. 2.42. Schematic of ESEM detect
Page 71 and 72:
Table 2.1. TECHNOLOGY LIMITING FACT
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3. DIGITAL IMAGES AND THE POSSIBILI
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Fig. 3.1. Image processing and anal
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Table 3.1. Comparison of selected f
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300 dpi, which corresponds to the p
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If we consider a recording element
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Fig. 3.4. Comparison of a circular
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Fig. 3.6. Changing depth of digital
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Fig. 3.7. Change in the appearance
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Fig. 3.8. Comparison of an image in
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Fig. 3.10. Determination of the num
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Maximum width This measurement can
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Fig. 3.11. Counting particles visib
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Let us analyse the distribution of
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An example of the application of co
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Fig. 3.16. The process of binarizat
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Image analysis software packages fr
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image is marked with the letter a).
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The effect of operation of these fi
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And another example of a filter: -1
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3.8. Mathematical morphology Morpho
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and the whole operation is repeated
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The simplest problem is presented i
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Fig. 3.29 illustrates the phenomeno
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Fig. 3.31. Model of RGB. Descriptio
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And substitution is an operation th
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increase decrease 255 contrast + 40
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Fig. 3.37. Application of normaliza
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4.1.2. Fixation of tissue structure
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The format most frequently used for
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Fig. 4.4. Histogram of grey levels
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orderline of two or three cells. Th
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Shape can also be described by mean
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Fig. 4.11. Image of the parenchyma
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4.2. Analysis of images obtained wi
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lens. However, the realization of t
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is now extremely easy, as all the w
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Fig. 4.16. Combination of erosion,
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Fig. 4.20. Division of binary image
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Fig. 4.24. Image labelling (AphImgC
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Fig. 4.28. Repeat watershed detecti
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as 60%), then the source of error i
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in natural state maintaining high s
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Therefore, the tasks of the compute
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Fig. 4.36. Result of Watershed oper
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and 5617 after correction. For carr
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Fig. 4.38. Histograms of cell area,
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should be analyzed separately. An e
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) Fig. 4.42. Result of reconstructi
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MICRO-STRUCTURE ANALYSIS OF PLANT TISSUES - Lublin

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