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Is the surface dirty? Are you looking for the clean surface or the dirt? If the dirt, no preparation is required! If the clean surface, dirt must be removed
Solvent in ultrasonic cleaner Methanol is best to finish cleaning Plasma cleaning also an option
Ultrasonic cleaner
Sample Preparation
Scanning Electron Microscope Beam of electrons rastered across sample Current flows from column to sample How does it complete the path back?
2. Variable pressure
1. Backfill gas bleeds off charge 2. Backfill gas amplifies signal 3. Allows imaging of non-conductive materials
Start simple
1. Conductive samples: metals, semiconductors
Fasten sample directly to stub Carbon tape: 8 mm to 50 mm wide x 20 m Carbon dots: 9 mm, 12 mm, 25 mm Carbon sheet: 5 x 12 cm Aluminum/Copper tape: single or double sided Silver paint Carbon paint
Carbon coating
High vacuum deposited: coating not conformal: works best on polished samples Simple thermal evaporation of carbon from yarn Preferred for analysis for light elements
Coatings can be THICK or THIN, particulate or smooth Coating are usually high Z metals such as Cr, Ta, W, Pt, Au With a THICK (20-50nm) coating the beam interaction occurs mainly within the coating Layer The SE-signal is then mostly SE2 (i.e. converted BS electrons) The topographic resolution is limited by the thickness of the metal coat and the SE II range (i.e ~mm)
SE II
Low Z material - polymer, or biological Low BSE coefficient Large exitation volume
SE II
There is very little SE II from the metal layer.. and no signal from specimen itself because SE produced beneath the metal layer cannot leave the specimen
Topographic resolution is now only limited by thickness of the metal coat and the diameter of the electron beam
Particulate Coatings
Au produces very big particles (30nm) so do not use pure gold for coatings Au/Pd, Pt, W, or Ir make smaller (<1-3nm) particles All have a very high SE yield and can be deposited in a low cost, sputter coater The coatings are stable for long periods of time Particulate coatings are ideal below 100kx but they can be useful even at higher magnifications..
Uncoated
Note the benefits of a reduction in charging and the gain in image contrast and detail. The fine grain - while visible - permits accurate focus and image stigmation. Resolution ~ 1.3nm
Quality Assurance
Particulate metal coatings Aim for a single layer coverage - thickness will then be equal to the average grain size, typically 1.5 to 3nm depending on the metal used Checking - the shadow on the filter paper should only be visible under strong, oblique, illumination and will be a blush of gray. Any hint of color, or a metallic sheen, indicates that the film is 1020nm thick at least
bulk value
1nm
2nm
3nm
Film thickness
If the metal layer is smooth and continuous then a new and important benefit occurs for high resolution imaging The SE1 varies with the thickness of the metal film This effect saturates at a thickness equal to about 3 The conformation of the film to surface topography thus provides contrast
50 nm
In this example a 1.5nm Cr film has been used The mass thickness contrast resolves edges and make nanometer scale detail (such as the molecular Fab fragments) readily visible The high SE yield also improves the signal to noise ratio of the image Unfortunately - use at once then discard because Cr films oxidize and degrade very rapidly in air
From Ren Hermann et al., (1991) J Struct Biol 107: 38-47
Depositing Cr films
Useful Cr coats cannot be deposited in a rough-pumped sputter coater because of the rapid oxidation that occurs The system should have a base pressure of 10-7 T and be dry pumped. After initial pump-down back fill the system with dry nitrogen to a pressure of about 1 Torr. Pump down once more to the base pressure, then back fill and pump again. Finally fill the liquid nitrogen trap to lower any residual water vapor pressure to below 10-8 T For the first 60 seconds of deposition the sample should be covered by a shutter to prevent oxide reaching the surface By experiment determine the slowest possible deposition rate. This ensures the smoothest and most uniform film
Structureless Coats
Structureless coatings Aim for 1 to 1.5nm to get the maximum SE1 contrast across features. These films are hard to focus and stigmate and intensity is low Checking the film thickness is impossible. Try varying the time until the best image is produced then copy exactly Remember that Cr, and Ti are very reactive and will oxidize rapidly even in a good vacuum. Once deposited these films should be used immediately and then discarded. Other materials may be more stable
Coating Summary
Coatings are an essential part of the technique of high resolution SEM because they generate interpretable contrast, improve resolution, and enhance the S/N ratio Thin coatings are better than thick coatings - do not make your sample a piece of jewelry Below 100kx magnification particulate coatings are superior to Cr or Ti because of their higher SE yields Above 100kx magnification can use chromium or titanium continuous films to generate mass thickness contrast and enhance resolution, or can use nano-grain Pt or W films (even though the grains are visible) for charge control and as an aid to accurate focus and stigmation Carbon is a contaminant not a coating unless it deposited by an ion sputter tool