Standard plant histology (Polemoniaceae)

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Method synopsis

Standard plant histological protocol involving glutaraldehyde fixation, resin embedding, serial sectioning at 1.5 μm, and multiple staining techniques including toluidine blue for general structure and specialized stains for cell wall components.

Synthesized from method descriptions across 1 paper 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 — Self-sterility in <i>Ipomopsis aggregata</i> (Polemoniaceae) is due to prezygotic ovule degeneration (2006), American Journal of Botany. Implementations in other papers (listed below) may differ.

1. Tissue fixation and processing
   Harvested carpels were immediately fixed in 3% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.2. Fixed tissues were dehydrated through an ethanol series, infiltrated with Spurr's resin, and polymerized at 70°C.
   Quantities: All harvested carpels processedDuration: Fixation immediate upon harvest; polymerization at 70°CConditions: Standard histological processing conditions with precise pH and temperature control
   Equipment: 3% glutaraldehyde, 0.1 M sodium cacodylate buffer, ethanol series, Spurr's resin
2. Serial sectioning
   Carpel tissue fixed in glutaraldehyde was sectioned at 1.5 μm thickness serially using standard microtomy techniques. Serial sections allowed complete examination of ovule and seed development patterns.
   Quantities: 1.5 μm thick serial sections of all processed carpelsDuration: Standard sectioning time per sampleConditions: Room temperature sectioning with sharp knives
   Equipment: microtome, 1.5 μm sectioning capability
3. Histological staining
   Sectioned tissues were stained with toluidine blue (O'Brien and McCully, 1981) to characterize general cellular structures, and counterstained with toluidine blue (O'Brien and McCully, 1981) to detect pectins. Some sections were stained with ether aniline blue to detect callose, or with phloroglucinol to detect lignin, or with ruthenium red to detect pectins.

Typical Equipment

• 3% glutaraldehyde
• 0.1 M sodium cacodylate buffer
• ethanol series
• Spurr's resin
• microtome
• toluidine blue
• ether aniline blue
• phloroglucinol
• ruthenium red

Output Measurements

• cellular structure
• cell wall composition
• tissue organization

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used