Critical steps and troubleshooting in sample preparation for wood and phloem formation : from sampling to microscopic observation

Balzano, Angela (author), Čufar, Katarina (author), Krže, Luka (author), Merela, Maks (author)

We present a technical note that supplements published procedures on optimal sample preparation for performing wood and phloem formation analyses. Before beginning sampling, it is important to learn about the characteristics of the tree or shrub species to be investigated. Some tips are given how to use the Trephor tool in the best way, how to remove the outer hard bark (periderm), how microcores should be handled after removal from the tree, and how they should be oriented for embedding in paraffin, and cutting thin sections for microscopy. Possible defects that may result from improper handling are illustrated and discussed. We also present optimal images to accurately identify different cell development stages in phloem and xylem, which is particularly challenging in hardwoods and Mediterranean tree and shrub species.

• Language: English
• Keywords: wood, tissue sampling, tissue preparation, microscopic slides, microscopy, xylogenesis, phloem formation, troubleshooting
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty
• Publication status: Published
• Publication version: Version of Record
• Publication date: 30.06.2022
• Year: 2022
• Number of pages: Str. 47-56
• Numbering: Let. 71, št. 1
• PID: 20.500.12556/RUL-138560
• UDC: 630*811.7:543.456
• ISSN on article: 0024-1067
• DOI: 10.26614/les-wood.2022.v71n01a05
• COBISS.SI-ID: 116719619
• Publication date in RUL: 28.07.2022

Record is a part of a journal

• Title: Les
• Shortened title: Les
• Publisher: Zveza društev inženirjev in tehnikov gozdarstva in lesarstva Slovenije, Zveza društev inženirjev in tehnikov gozdarstva in lesarstva Slovenije, Biotehniška fakulteta, Oddelek za lesarstvo, Založba Univerze, Biotehniška fakulteta, Oddelek za lesarstvo, Založba Univerze
• ISSN: 0024-1067
• COBISS.SI-ID: 13940224

Licences

• License: CC BY-NC 4.0, Creative Commons Attribution-NonCommercial 4.0 International
• Link: http://creativecommons.org/licenses/by-nc/4.0/
• Description: A creative commons license that bans commercial use, but the users don’t have to license their derivative works on the same terms.
• Licensing start date: 28.07.2022

Secondary language

• Language: Slovenian
• Title: Kritični koraki in reševanje težav pri pripravi vzorcev za spremljanje nastajanja lesa in floema : od vzorčenja do opazovanja pod mikroskopom

Abstract

Predstavljamo tehnična navodila, ki dopolnjujejo objavljene postopke za optimalno pripravo vzorcev za izvajanje analiz nastajanja lesa in floema. Pred začetkom vzorčenja je potrebno poznati značilnosti drevesne ali grmovne vrste, ki jo želimo preiskovati. Podanih je nekaj nasvetov, kako najbolje uporabiti orodje Trephor, kako odstraniti zunanjo trdo skorjo (periderm), kako ravnati z mikro izvrtki po odvzemu iz debla in kako jih orientirati za vklapljanje v parafin in rezanje tankih rezin za mikroskopske preiskave. Prikazane in razložene so možne napake, ki lahko nastanejo zaradi neustreznega ravnanja v različnih korakih postopka. Predstavljamo tudi optimalne slike tkiv za prepoznavanje različnih razvojnih faz celic v floemu in ksilemu, ki je še posebej zahtevno pri listavcih ter sredozemskih drevesnih in grmovnih vrstah.

• Keywords: les, odvzem vzorcev tkiva, priprava vzorcev, prostorninsko krčenje, mikroskopski preparati, mikroskopiranje, nastajanje lesa, nastajanje floema, odpravljanje težav

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: P4-0015
• Name: Les in lignocelulozni kompoziti