Scanning Electron Microscopy Project Portfolio
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1 Scanning Electron Microscopy Project Portfolio Prepared by: Submitted for: CME 596 Scanning Electron Microscopy Fall 2015 N.C. Brown Center for ultrastructure Studies
2 Part I A portfolio of micrographs exhibiting the following techniques: Critical Point Drying Depth of Field Backscatter Low Voltage Image of an Uncoated Sample High Magnification (>50,000x) Stereo Pair Cryofracture
3 Critical Point Drying Figure 1. Scanning electron micrograph of truffle fungal spores in a hyphal mass Magnification: 1900x Voltage: 7kV Working Distance: 13 Spot Size: 11 Chevrons Aperture: 1
4 Depth of Field Figure 2a. Scanning electron micrograph of truffle surface Magnification: 190x Voltage: 5kV Working Distance: 7 Spot Size: 12 Chevrons Aperture: 1 Tilt: +38.4
5 Depth of Field Figure 2b. Scanning electron micrograph of truffle surface Magnification: 600x Voltage: 5kV Working Distance: 22 Spot Size: 11 Chevrons Aperture: 1 Tilt: +29.7
6 Backscatter Figure 3a. Backscatter scanning electron micrograph of Chromium, an atomic number of 24 (left). and Tungsten, an atomic weight of 74 (right) Magnification: 800x Voltage: 20kV Working Distance: 13 Spot Size: 16 Chevrons Aperture: 2 Figure 3b. Secondary scanning micrograph of Chromium, Cr (left), and Tungsten, W (right) Magnification: 800x Voltage: 20kV Working Distance: 13 Spot Size: 16 Chevrons Aperture: 2
7 Low Voltage Image of an Uncoated Sample Figure 4. Scanning electron micrograph of Vanadium Magnification: 1200x Voltage: 1.8kV Working Distance: 12 Spot Size: 11 Chevrons Aperture: 1
8 High Magnification Figure 5. Scanning electron micrograph of the facial hair of a Wasp. A second sputter coating of the specimen should prove to reduce the charging at high magnification Magnification: 55000x Voltage: 5kV Working Distance: 13 Spot Size: 8 Chevrons Aperture:1
9 Stereo Pair Figure 6. Scanning electron micrograph stereo pair of truffle hyphae Magnification: 500x Voltage: 5kV Working Distance: 13 Spot Size: 12 Chevrons Aperture: 1 Tilt: -7.9
10 Cryofracture Figure 7. Scanning electron micrograph of a cryo-fractured oak leaf stem, looking down the stem Magnification: 1900x Voltage: 7kV Working Distance: 11 Spot Size: 12 Chevrons Aperture 1
11 Part II Biological Sample: A series of micrographs relating to a biological specimen Fungal Specimens fixed in paraformaldehyde (4%), gluteraldehyde (0.1%) in PBS buffer and stained with osmium tetroxide (1% in water). Specimens were then critically point dried, placed on an aluminum stub with carbon paint and sputter coated. Insect specimens were placed on an aluminum stub and sputter coated. Non-biological: A series of micrographs relating to a non-biological specimen Paperclip specimens were uncoated and mounted on an aluminum stub with carbon paint Thread was sputter coated and mounted on an aluminum stub with carbon paint Vanadium (Vn) was uncoated and mounted on an aluminum stub with carbon paint
12 Biological Specimen Figure 8. Scanning electron micrograph of a truffle spore attached to hyphae. The spores are located on the inside of the truffle. This is a cross-sectioned image. Magnification: 2300x Voltage: 7kV Working Distance: 13 Spot Size: 10 Chevrons Aperture 1
13 Biological Specimen Figure 9. Scanning electron micrograph of a truffle spore attached to hyphae. Some charging can be seen on the left hand side of the image and was decreased by double coating the specimen Magnification: 3500x Voltage: 7kV Working Distance: 13 Spot Size: 10 Chevrons Aperture 1
14 Biological Specimen Figure 10. Scanning electron micrograph of a truffle spore attached to hyphae. Charging was corrected by as second sputter coat Magnification: 5500x Voltage: 7kV Working Distance: 13 Spot Size: 10 Chevrons Aperture: 1
15 Biological Specimen Figure 11. Scanning electron micrograph of a broken wasp mouth mandible Magnification: 900x Voltage: 8kV Working Distance: 12 Spot Size: 12 Chevrons Aperture 1
16 Biological Specimen Figure 12. Scanning electron micrograph of a broken wasp mouth mandible Magnification: 1800x Voltage: 8kV Working Distance: 12 Spot Size: 12 Chevrons Aperture 1
17 Non-Biological Specimen Figure 13. Scanning electron micrograph of an uncoated paperclip Magnification: 1800x Voltage: 8kV Working Distance: 12 Spot Size: 12 Chevrons Aperture 1
18 Non-Biological Specimen Figure 14. Scanning electron micrograph of an uncoated paperclip. The marks seen here are indentations from a pair of pliers used to break the paperclip. Magnification: 300x Voltage: 8kV Working Distance: 14 Spot Size: 13 Chevrons Aperture 1
19 Non-Biological Specimen Figure 15. Scanning electron micrograph of a sputter coated mass of sweater thread. Some charging can be seen where the coating was too thin. Magnification: 1800x Voltage: 8kV Working Distance: 12 Spot Size: 12 Chevrons Aperture 1
20 Non-Biological Specimen Figure 16. Scanning electron micrograph of an uncoated sample of Vanadium (Vn). Magnification: 1600x Voltage: 1.8kV Working Distance: 12 Spot Size: 11 Chevrons Aperture 1
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