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Figure 6.13: 3-D view of the middle part of a dried E. gracilis cell. The arrows point towards features clearly identified with pellicle pores, as they have been known from SEM investigations since the late 1960s ([116]). The arrowheads indicate new surface features that seems to have no correlation in other imaging data of E. gracilis such as acquired by SEM and TEM. Intermittent contact mode AFM data, 3D-view based on Height trace. Figure 6.14: A SEM image of the pellicle of the alga E. terricola. The pellicle pores as indicated by the arrows are distributed along the ridges, creating small narrowings of the adjacent pellicular strips. Indentation features like these on the Euglena pellicle that appear centered on the strips are not to be found. Scale bar is 2 µm. Image adapted from [116]. 88
Crystalline Cell Parts Crystalline cell parts had to be imaged after a series of preparation and purification steps. Successful imaging included semicrystalline starch deposits (paramylum grains) and cell parts. AFM imaging of such a cell part solution has never been done before and in this sense the next images are unique. A view on a dried sample by optical microscopy is seen in Fig. 6.15. Small lumps of material aggregated and it often was difficult to locate intact cell parts within. In comparison, in Fig. 6.16 a patch of dried crystalline cell part solution as imaged by AFM is shown. This patch is of good quality because it does not only contain the residue of dissolved biological material but also crystalline cell parts. E.g. a paramylum grain can be spotted to the lower left of the image. In Fig. 6.17 a feature often found in such a dried crystalline cell parts preparation is seen. The size and shape of this feature correlate to those of a lipid body, one of Euglena’s energy storage facilities. The search for the photoreceptor of E. gracilis yielded the imaging of very interesting regular structures such as the one in Fig. 6.18. Despite the layered nature of the structure, further AFM investigation is needed for a more complete assessment. An E. gracilis cell contains only one photoreceptor at a mass of only about 1/20000th of that of the whole cell. Even in our purified solution a targeted approach using AFM is difficult. The last figure, Fig. 6.19, gives a 3D-view of a paramylum grain based on AFM height information. It confirms the overall concentric pattern found in such a grain stemming from the crystalline assembly of microfibrils only 4 nm in diameter. On page 60, Fig. 4.18 shows a SEM image of a freeze-fractured paramylum grain. The details of the microfibril assembly are not yet fully understood. ([108], [113]) Figure 6.15: The cantilever scanning over a dried solution of crystalline cell parts. Image taken by optical light microscopy (transmitted light). 89
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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
Page 37 and 38: Figure 3.14: This force curve shows
Page 39 and 40: 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: Figure 6.11: The pellicle of an E.
Page 91 and 92: Figure 6.18: A layered structure th
Page 93 and 94: It might also be possible that comb
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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