Acrylic leaf imprint stomatal analysis

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Method synopsis

Acrylic imprints of leaf surfaces are made on glass slides and analyzed under compound microscope to count stomata and measure their diameters.

Synthesized from method descriptions across 2 papers using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Plasticity in functional traits in the context of climate change: A case study of the subalpine forb Boechera stricta (Brassicaceae) (2015), Global Change Biology. Implementations in other papers (listed below) may differ.

1. Process leaf samples for morphological measurements
   Scanned fresh leaves using ImageJ software, dried at 50°C for 3 days, then weighed. Calculated specific leaf area (SLA) as leaf area/dried weight. Used 528 individuals for SLA measurements. Prepared epidermal impressions by mounting dried leaves on microscope slides using clear nail polish to create impressions of abaxial leaf surface.
   Quantities: 528 individuals for SLA, 4 distinct non-overlapping 0.0352 mm² areas photographed per impression at 400x magnificationDuration: 3 days drying time at 50°CConditions: Laboratory processing
   Equipment: ImageJ software, microscope, microscope slides, clear nail polish, drying oven, analytical balance
2. Measure stomatal characteristics
   Photographed 4 distinct non-overlapping 0.0352 mm² areas of each epidermal impression under 400x magnification. Averaged stomatal density over fields, then used ImageJ to quantify stomatal size by measuring distance between two inner dimensions of guard cells for 6 stomata per plant.
   Quantities: 4 fields per impression, 6 stomata measured per plant, 0.0352 mm² area per field, 400x magnificationDuration: Microscopy sessions during 2012-2013Conditions: Laboratory conditions
   Equipment: compound microscope, ImageJ software, digital camera

Typical Equipment

• acrylic
• glass slides
• clear tape
• compound microscope
• 10x/0.25 objective
• 100x/1.25 eyepiece
• 10x10 grid eyepiece

Output Measurements

• stomatal density (stomata/cm²)
• stomatal diameter (μm)

Papers Using This Protocol (2)

Species Studied With This Protocol

Places Where Used