Cell surface and cell outline imaging in plant tissues using the backscattered electron detector in a variable...

Area of Science:

• Plant biology
• Microscopy techniques
• Image analysis

Background:

• Scanning electron microscopy (SEM) is a long-standing technique for high-resolution plant cell surface imaging.
• Traditional SEM requires metal coating of samples to prevent charging, limiting analysis of uncoated tissues.
• Variable pressure-SEM (VP-SEM) enables imaging of uncoated plant tissues using secondary or backscattered electron detectors.

Purpose of the Study:

• To outline methods for backscattered electron imaging of diverse plant tissues.
• To demonstrate the advantages of backscattered electron imaging for high-contrast cell outlines.
• To provide a protocol for imaging and quantifying Arabidopsis thaliana leaf epidermal cells.

Main Methods:

• Utilized backscattered electron (BSE) detector under low vacuum conditions.
• Imaged uncoated barley leaf and Arabidopsis thaliana leaf epidermal tissues.
• Optimized imaging parameters like accelerating voltage and chamber pressure for various plant tissues.

Main Results:

• Achieved high-contrast cell outlines suitable for semi-automated image analysis.
• Demonstrated effective imaging of uncoated plant morphology and cell structures.
• Successfully quantified cell areas in barley leaf tissue using BSE imaging.

Conclusions:

• Backscattered electron imaging of uncoated plant tissue provides detailed morphology and clear cell outlines.
• The method is adaptable to various plant tissues and requires standard SEM equipment.
• This technique is suitable for laboratories with variable pressure-SEM or tabletop SEM.