Conventional SEM Specimen at high vacuum – requires sample fixation and dehydration or freezing. Charging is minimized by coating sample with metal or carbon or lowering the operating kV.
Variable Pressure Scanning Electron Microscope -Vacuum in the sample chamber can range from high vacuum (< 10-6 Pascals) up to 3,000 Pa. -Environmental- 3,000 Pa/ Variable – 750 Pa - Gas in the sample chamber allows uncoated and unfixed samples to be imaged. -Detectors used at higher pressures are backscatter or special secondary detectors. - Moisture on the sample can be controlled by cooling/heating stage and water injection system.
LV SEM – High Vacuum Mode
LV SEM - Low Vacuum Mode
SEM Electron/Specimen Interactions When the electron beam strikes a sample, both photon and electron signals are emitted. Incident Beam Specimen X-rays - composition info Auger electrons - Surface sensitive compositional Backscattered electrons - Atomic number and topographical Cathodoluminescence - Electrical Secondary electrons - Topographical Specimen Current Electrical
Secondary Electron Detector
Backscatter electron detector
Signal Detection in Variable Pressure Mode
Charging is minimized
Peltier stage Heats to 50 C Cools to - 25 C
Control of Water vapor and temperature
Applications Live centipede Bacteria on rock
Fresh moss with liquid water
Pattern produced in silica gel
Live Drosophila larva