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Chapter 7 Outlook The Atomic Force Microscope (AFM) has proven to be a valuable tool for investigation of biomaterials at the small scale. In this work the single celled alga Euglena gracilis was investigated in the light of being a bionanotechnological system accommodating a wealth of functional units within only small volume. A method for AFM data acquisition of the alga Euglena gracilis was developed and further investigation can build upon this approach. As this work is only a beginning of the AFM study of this alga, it focused on its morphology, on how to obtain first image data of Euglena’s pellicle and some of its functional cell organelles with potential for technical applications. This undertaking can be seen as an ignition part in the investigation of algal materials exhibiting molecular precision achitecture and complexity inherent to living systems. While the data obtained on whole cells as well as cell organelles provided essentially morphological data and proved the feasibility of such studies by means of AFM, other analytical characterization methods, e.g AFM force spectroscopy, are now possible on these and similar algae by making use of the preparation method developed herein. Since AFM yields information about the sample as well as it allows manipulation on the micro and nanoscale, future research attempts along these lines seem encouraging. The exact nature of the novel features found, i.e. the indentations in the center of pellicular strips, should be further analyzed, especially with regards to the sample preparation techniques used. Equally interesting, AFM force spectroscopy shall characterize the mucus material ejected by mucus excretion pellicle pores. Further, elucidation about the three-dimensional structure of the photoreceptor was attempted with this work, however, the fraction of crystalline parts contained too few photoreceptors to be found with the AFM tip, since crystalloid bodies, paramylon grains and not completely removed cell material amounted for a large portion of the fraction. Possible future approaches comprise the development of a preparation technique for a solution containing a plentitude of photoreceptors. 92
It might also be possible that combination with fluorescence microscopy simplifies observational search for the location of the photoreceptor. As bottomline, a number of future research directions seem worthwhile. AFM can help relating structure to function in biomaterials, a precursor to develop bioinspired artifial systems. Euglena gracilis is an ideal object of study when general nanotechnological design principles of living systems are to be investigated. ([119]) 93
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DIPLOMARBEIT Atomic Force Microscop
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»Later I have been asked many time
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Contents Abstract 4 1 Introduction
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5.2.1 Optical Microscopy and Fluore
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theory (see e.g. [1]). Now I call o
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understand phenomena at a larger sc
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of whole dried Euglena cells in air
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these assembly functions for us. 2
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Figure 2.2: Rise in public spending
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The presented three types of nanote
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2.4.1 Self-Assembly and Emergent St
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After diligent investigation such s
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1 µN) between a probe and the samp
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3.2.1 Static Mode In Static Mode (a
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crystal and the cantilever tip (pha
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Figure 3.6: Highly ordered and fres
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Figure 3.8: Schematic drawing of th
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Figure 3.11: The cantilever deflect
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Figure 3.14: This force curve shows
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Figure 4.1: A group of Euglena orga
Page 41 and 42: several species with rigid pellicle
Page 43 and 44: Figure 4.2: The rightmost cell can
Page 45 and 46: cavity and is used for regulatory f
Page 47 and 48: as phobic reactions, because the ce
Page 49 and 50: Figure 4.7: Scheme of the photorece
Page 51 and 52: Figure 4.9: Measured monoclinic uni
Page 53 and 54: Figure 4.11: Twice the same Euglena
Page 55 and 56: 4.15), their ends are termed the (-
Page 57 and 58: Figure 4.15: Kinesin and dynein and
Page 59 and 60: Figure 4.17: (Image reproduced from
Page 61 and 62: into fully functional chloroplasts
Page 63 and 64: Figure 4.19: How a microtubule-kine
Page 65 and 66: Chapter 5 Materials and Methods The
Page 67 and 68: Cell Treatment - Demembranation In
Page 69 and 70: Figure 5.3: This image shows cells
Page 71 and 72: Figure 5.4: The AFM setup at the In
Page 73 and 74: Figure 5.6: If the user menu AFM_TU
Page 75 and 76: Figure 5.7: Single steps of the sam
Page 77 and 78: Chapter 6 Results High resolution t
Page 79 and 80: Figure 6.2: An embedded E. gracilis
Page 81 and 82: Figure 6.5: Scanning under liquid s
Page 83 and 84: Flagellum The flagellum of an E. gr
Page 85 and 86: Pellicle The pellicle of E. gracili
Page 87 and 88: Figure 6.11: The pellicle of an E.
Page 89 and 90: Crystalline Cell Parts Crystalline
Page 91: Figure 6.18: A layered structure th
Page 95 and 96: Appendix A Acronyms SPM Scanning Pr
Page 97 and 98: [13] A. Junk, F. Riess: From an ide
Page 99 and 100: [45] J. Ruan, B. Bhushan: Atomic-sc
Page 101 and 102: [70] M. Rief, M. Gautel, F. Oesterh
Page 103 and 104: [97] P. Gualtieri, L. Barsanti: Alg
Page 105 and 106: Annex 1 This Igor script was used i
Page 107 and 108: w[i][j][laynr]=(w[i][j][3]-used_off
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