Synchrotron X-ray leaf venation imaging (Plantae)

Knowledge graph centered on Synchrotron X-ray leaf venation imaging (Plantae) with 19 nodes and 45 connections. Top connected: Asteraceae, Arabidopsis, X-ray imaging of leaf venation networks, Arabidopsis thaliana, fish and wildlife.

Method synopsis

High-resolution X-ray imaging of leaf venation networks using synchrotron light sources with both absorption and phase contrast approaches. Samples are mounted on translation stages and imaged with monochromatic X-rays at various energies and detector distances.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure as described in the canonical source

Steps below were extracted from the paper that introduces this protocol — X-ray imaging of leaf venation networks (2012), New Phytologist. Implementations in other papers (listed below) may differ.

1. Collect and prepare leaf samples
   Collected 408 leaves from 44 woody and herbaceous species spanning a wide climate gradient in the Colorado Rocky Mountains. Each leaf was pressed flat and dried at 60°C for at least 3 days, after which a 5 × 5 mm² section was cut from the lamina of each leaf using a razor blade.
   Quantities: 408 leaves from 44 species for synchrotron absorption contrast experiments; 599 leaves from 37 species for iodine contrast experimentsDuration: At least 3 days drying time at 60°CConditions: Leaves dried at 60°C
   Equipment: razor blade
2. Mount samples for synchrotron imaging
   Each sample was mounted on small aluminum platform using modeling clay; these platforms were then rapidly transferred by robotic arm to a translation stage placed in the beam path. X-rays were converted to visible light using a lutetium aluminum garnet (LuAG) scintillation crystal and then imaged using a 4× microscope objective and 2048 × 2048 pixel 12-bit CCD array.
   Quantities: 2048 × 2048 pixel array, 12-bit resolution, final resolution was 1.66 μm pixel⁻¹Duration: Maximum exposure time of 450 ms per imageConditions: Under 2 d beamline time at APS's 2-BM beamline
   Equipment: aluminum platform, modeling clay, robotic arm, translation stage, lutetium aluminum garnet (LuAG) scintillation crystal, 4× microscope objective, 2048 × 2048 pixel 12-bit CCD array
3. Perform synchrotron X-ray imaging
   Fixed L at 55 m while varying d from 0.01 to 0.10 m, and varied the X-ray energy from 7 to 30 keV. For each sample a bright field (with sample), flat field (no sample), and dark field (no X-rays) were acquired with a maximum exposure time of 450 ms. A final image was calculated as (bright – dark)/(flat – dark) and then local contrast was further enhanced using contrast-limited adaptive histogram equalization.

Typical Equipment

• aluminum platform
• modeling clay
• robotic arm
• translation stage
• lutetium aluminum garnet (LuAG) scintillation crystal
• 4× microscope objective
• 2048 × 2048 pixel 12-bit CCD array
• bending magnet
• double-bounce multi-layer monochromator

Output Measurements

• venation network structure
• vein connectivity
• spatial resolution images

Papers Using This Protocol (1)

Species Studied With This Protocol