Automation of Scanning Electron Microscopy Sample Prep

Automating SEM sample prep involves using specialized technologies and systems to streamline the process:

• • Fixation: Automated systems apply chemical fixatives (e.g., glutaraldehyde) to preserve sample structure.
  • Rinsing:Robots rinse samples in buffer solutions to remove excess fixatives.
  • Dehydration: Automated solvent exchange systems progressively replace water with ethanol or acetone through a graded series.
  • Drying:Critical point drying or automated freeze-drying systems remove all remaining liquid without collapsing fine structures.
  • Mounting: Robotic arms or precision tools place samples on SEM stubs or holders using conductive adhesives or tapes.
  • Sputter Coating: An automated sputter coater deposits a thin, uniform layer of conductive material (e.g., gold, platinum) on the sample surface.
  • Labelling and Tracking: Systems integrate barcode or RFID tagging for sample identification and tracking.

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Benefits of Automation Over Manual Methods

• Increased Speed:Automated systems significantly speed up the sample preparation process, allowing for higher throughput and faster turnaround times, which is especially valuable in large-scale studies.
• Improved Reproducibility: Automation ensures consistent sample preparation by minimizing human error and variability between different samples or operators, leading to more reliable results.
• Higher Throughput: Automation allows for handling larger sample volumes simultaneously, ideal for high-throughput projects.
• Better Accuracy and Precision: Robots perform tasks with higher precision, reducing contamination and errors.
• Cost Savings: Though the upfront cost can be higher than manual methods, automation reduces labor and reagent waste in the long run.

Methods of Scanning Electron Microscopy (SEM) Sample Prep

Chemical Fixation 

Preserves biological specimens by crosslinking proteins using agents like glutaraldehyde and osmium tetroxide.

Critical Point Drying

Replaces water in samples with a transitional fluid, then dries it under specific temperature and pressure to avoid surface tension damage.

Sputter Coating

Deposits a thin conductive metal layer (e.g., gold, palladium) on non-conductive samples to prevent charging under the electron beam.

Freeze Fracturing

Rapidly freezes and fractures samples to expose internal structures, often followed by shadowing with metal for imaging.