Transmission Electron Microscopy of the Phloem with Minimal Artifacts
Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
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Transmission Electron Microscopy of the Phloem with Minimal Artifacts. / Hunziker, Pascal; Schulz, Alexander.
Phloem: Methods and Protocols. red. / Johannes Liesche. Humana Press, 2019. s. 17-27 (Methods in Molecular Biology, Bind 2014).Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
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TY - CHAP
T1 - Transmission Electron Microscopy of the Phloem with Minimal Artifacts
AU - Hunziker, Pascal
AU - Schulz, Alexander
PY - 2019
Y1 - 2019
N2 - It is a universal feature of seed plants that their phloem consists of a continuous sieve-tube system throughout the plant that is highly pressurized by its sugar contents. Cellular continuity and the pressure flow, osmotically generated in the source leaves, allow the assimilates to reach all sinks organs. However, both phloem features, the cellular continuity and the high pressure, are challenges when fixing the phloem for transmission electron microscopy. With very few exceptions, the tissue preparation necessary for the fixation evokes rapid wound responses that eventually result in artifacts. This chapter describes the steps necessary to minimize development of artifacts in the phloem and includes preparation of fixatives, a dissection procedure that optimizes penetration of the fixatives and application to axial and lateral plant organs. Moreover, as alternative to the established fixation of fresh hand sections, we suggest a xylem-assisted perfusion fixation method for herbaceous plants. After the initial fixation, the subsequent dehydration, embedding, and ultrathin sectioning of the material follow routine procedures, which are briefly discussed, as is the orientation of samples for obtaining transverse and longitudinal phloem sections.
AB - It is a universal feature of seed plants that their phloem consists of a continuous sieve-tube system throughout the plant that is highly pressurized by its sugar contents. Cellular continuity and the pressure flow, osmotically generated in the source leaves, allow the assimilates to reach all sinks organs. However, both phloem features, the cellular continuity and the high pressure, are challenges when fixing the phloem for transmission electron microscopy. With very few exceptions, the tissue preparation necessary for the fixation evokes rapid wound responses that eventually result in artifacts. This chapter describes the steps necessary to minimize development of artifacts in the phloem and includes preparation of fixatives, a dissection procedure that optimizes penetration of the fixatives and application to axial and lateral plant organs. Moreover, as alternative to the established fixation of fresh hand sections, we suggest a xylem-assisted perfusion fixation method for herbaceous plants. After the initial fixation, the subsequent dehydration, embedding, and ultrathin sectioning of the material follow routine procedures, which are briefly discussed, as is the orientation of samples for obtaining transverse and longitudinal phloem sections.
KW - Aldehyde fixatives
KW - Companion cell
KW - Perfusion fixation
KW - Phloem fixation
KW - Pressure release
KW - Sieve element
KW - Transmission electron microscopy
KW - Wound response
U2 - 10.1007/978-1-4939-9562-2_2
DO - 10.1007/978-1-4939-9562-2_2
M3 - Book chapter
C2 - 31197783
AN - SCOPUS:85067430560
SN - 978-1-4939-9561-5
T3 - Methods in Molecular Biology
SP - 17
EP - 27
BT - Phloem
A2 - Liesche, Johannes
PB - Humana Press
ER -
ID: 224335990