Scanning Electron Microscopy Laboratory Portfolio
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1 SUNY College of Environmental Science and Forestry Digital ESF N.C. Brown Center for Ultrastructure Studies Fall 2016 Scanning Electron Microscopy Laboratory Portfolio Marissa Lanzatella SUNY College of Environmental Science and Forestry Follow this and additional works at: Part of the Nanoscience and Nanotechnology Commons, and the Structural Materials Commons Recommended Citation Lanzatella, Marissa, "Scanning Electron Microscopy Laboratory Portfolio" (2016). N.C. Brown Center for Ultrastructure Studies This Presentation is brought to you for free and open access by Digital ESF. It has been accepted for inclusion in N.C. Brown Center for Ultrastructure Studies by an authorized administrator of Digital ESF. For more information, please contact digitalcommons@esf.edu.
2 Scanning Electron Microscopy Laboratory Portfolio Marissa Lanzatella December 5, 2016 Submitted for MCR 484/783 Scanning Electron Microscopy Fall 2016 N.C. Brown Center for Ultrastructure Studies
3 These images were prepared as part of the class MCR 484 Scanning Electron Microscopy at SUNY College of Environmental Science and Forestry, Fall 2016 All images were acquired on the JEOL JSM 5800 LV Scanning Electron Microscope in the N. C. Brown Center for Ultrastructure Studies 2
4 Marissa Lanzatella Major: Biotechnology Minor: Microscopy Career Goals: to research mycology utilizing microscopy The images found in this collection are examples of the knowledge and skills I have developed through the MCR 484 Scanning Electron Microscopy course taken in the fall of I took this course because I hope to work as a microscopist in the future. Electron microscopy is of particular interest to me. I wanted to gain as much experience as possible on various pieces of equipment, such as the SEM. 3
5 Table of Contents A portfolio of micrographs from lab sessions demonstrating the following techniques: 1. Best Image 2. Most Difficult Image 3. Favorite Image 4. Standard Low Magnification 5. Secondary Electron Image and Probe diameter a. Spot size 8 b. Spot size Specimen Preparation: Propylene Oxide 7. Specimen Preparation: Sputter Coating 8. Specimen Preparation: Critical Point Drying 9. Image Quality I: Depth of Field 10. Image Quality II: Accelerating Voltage 11. Backscattered Electron Imaging a. SEI image b. BEI image c. BEI image 12. Low voltage of Uncoated Biological Sample The images I am presenting in this collection were chosen because they exemplify the knowledge and skills I have developed along with the care, quality, and concern for the work I produce. 4
6 Figure 1: Best Image I have chosen this as my best image because it is sharply focused and has a decent depth of field, though towards the top edge some features are lost. It is also very interesting to look at, with the sharp angle of the crystal and encompassing rock. Its pair, a BEI image, gives a unique perspective but is unfortunately not in focus as well. 5
7 Figure 1. One of two images comparing SEI and BEI using a sample of a geode. This image features SEI. It has a partial depth of field. The top is out of focus. Accelerating Voltage 20 kv; Spot Size 16; Objective Aperture 2; Working Distance 15 mm; 500x magnification; 20 6 micrometer bar; SEM.
8 Figure 2: Most Difficult Image I have chosen this as the hardest image to capture because of the multiple attempts at image processing it took. It shows my progress by the end of the semester. It reveals not only my processing abilities but also my basic competency with the SEM. The image still could be improved; my strength with the Z control was not the greatest and I should have rotated the image slightly more to ensure they overlapped better. 7
9 Figure 2. Stereoimage of sputtercoated carbonized wood. Depth can be seen around the edges; the two ascending wood fragments did not match their images well after being tilted, as seen by the discrepancies in green and red. Accelerating Voltage 15 kv; Spot Size 8; Objective Aperture 1; Working Distance 35 mm; 200x magnification; 50 micrometer bar; SEM. 8
10 Figure 3: My Favorite Image I have chosen this as my favorite because it was my first micrograph and I am proud of it. No special techniques were employed for it but looking at this image inspires feelings of triumph in me as I think of how much I have learned and that I would really like to pursue microscopy. Of course, the charging could be reduced and some information was lost compared to a smaller spot size. The depth of field also needs improvement. 9
11 Figure 3. One of two images comparing spot size using the eye of a bark beetle. The larger spot size displays more charging, less graininess, and the loss of some information. Accelerating Voltage 15 kv; Spot Size 16; Objective Aperture 2; Working Distance 14 mm; 650x magnification; 20 micrometer bar; 10 SEM.
12 Additional Examples of My Work The following images are additional examples of my work; I have included them because they show my progress as I learned more about the SEM and my capability with certain techniques. 11
13 Figure 4. Low magnification image of a coated mite s mouthparts. Accelerating voltage was decreased to reduce charging. Charging built up on fine hairs. Quality is much better than at the higher magnifications. Accelerating Voltage 15 kv; Spot Size 9; Objective Aperture 1; Working 12 Distance 14 mm; 1900x magnification; 5 micrometer bar; SEM.
14 Figure 5a. One of two images comparing spot size using the eye of a bark beetle. The smaller spot size displays more graininess but less charging. Accelerating Voltage 15 kv; Spot Size 8; Objective Aperture 2; Working Distance 14 mm; 650x magnification; 20 micrometer bar; SEM. 13
15 Figure 5b. One of two images comparing spot size using the eye of a bark beetle. The larger spot size displays more charging, less graininess, and the loss of some information. Accelerating Voltage 15 kv; Spot Size 16; Objective Aperture 2; Working Distance 14 mm; 650x magnification; 20 micrometer 14 bar; SEM.
16 Figure 6. A micrograph of a maple leaf treated with propylene oxide. The cellular debris is a result of cell lysis due to the treatment. The leaf vein sustained some damage but was hardier than the surrounding cells. Accelerating Voltage 15 kv; Spot Size 9; Objective Aperture 2; Working Distance mm; 1300x magnification; 10 micrometer bar; SEM.
17 Figure 7. The sputtercoated underside of a succulent leaf. It was sputtercoated for 45 seconds with Au/Pd at 45 kv. The central structure is unknown. Accelerating Voltage 15 kv; Spot Size 8; Objective Aperture 2; Working Distance 20 mm; 600x magnification; 20 micrometer bar; SEM. 16
18 Figure 8. The top layer of a critical point dried leaf. The raised lines are veins and the central object is likely a salt crystal. Accelerating Voltage 15 kv; Spot Size 9; Objective Aperture 2; 17 Working Distance 20 mm; 2000x magnification; 5 micrometer bar; SEM.
19 Figure 9. Depth of field analysis at a medium aperture and short working distance of a TEM grid. One of four images showing different comparisons of aperture and working distance. While a lot of detail is visible, the very edges of the grid do not have lines as crisp as the center. Accelerating Voltage 10 kv; Spot Size 16; Objective Aperture 2; Working Distance 12 mm; 200x Magnification; 50 micrometer bar; 18 SEM.
20 Figure 10. Watch screw surface at a low accelerating voltage. One of two images to compare accelerating voltage effects. It has a decent depth of field but is grainy. Accelerating Voltage 10 kv; Spot Size 13; Objective Aperture 1; Working Distance 20 nm; 2700x Magnification; 5 19 micrometer bar; SEM.
21 Figure11a. One of two images comparing SEI and BEI using a sample of a geode. This image features SEI. It has a partial depth of field. The top is out of focus. Accelerating Voltage 20 kv; Spot Size 16; Objective Aperture 2; Working Distance 15 mm; 500x magnification; micrometer bar; SEM.
22 Figure 11b. One of two images comparing SEI and BEI using a sample of a geode. This image features BEI. It has a larger area out of focus. The details aren t as strong. Accelerating Voltage 20 kv; Spot Size 16; Objective Aperture 2; Working Distance 15 mm; 500x magnification; micrometer bar; SEM.
23 Figure 11c. Molybdenum ore viewed under BEI. Some solid carbon, or other element of low atomic number, appears to also be present as the darker crystals. Accelerating Voltage 20 kv; Spot Size 14; Objective Aperture 2; Working Distance 16 mm; 500x magnification; 20 micrometer bar; SEM. 22
24 Figure 12. Low voltage image of uncoated monkey hair. Stage was tilted. Some detail is apparent but the lines are not all crisp. Accelerating Voltage.8 kv; Spot Size 15; Objective Aperture 2; Working Distance 25 mm; 550x magnification; 20 micrometer bar; SEM. 23
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