Fanny Hidayati, Ph.

D
 Microscopic Examination of Wood: Sample
Preparation and Techniques for Light
Microscopy by Jacques C. Tardif and France
Conciatori IN Plant Microtechniques and
Protocols with editors: Edward Chee Tak
Yeung, Claudio Stasolla, Michael John Sumner,
Bing Quan Huang
 Penerbit: Springer
 Tahun 2015
 Stained, dehydrated, and cleared thin
sections will be permanently mounted using
undamaged and clean, dry slides and
coverslips.
 We usually mount the best 2–3 sections per
slide from the initial pool of thin sections to
assure that backup sections are available.
 If transversal, radial, and tangential sections are
to be mounted on a single slide,the staine d
sections are transferred from the watch glass to
the mounting slide immediately after the clearing
stage.
 A fine brush may be used to transfer the section
but we prefer using fine tip Dumoxel tweezers.
 The excess clearing solvent (e.g., xylene, d-
limonene) is drained using absorbent paper to
avoid unnecessary thinning of the mounting
medium.
 At this stage, the practitioner should work
quickly (and with confidence) to avoid letting the
sections dry on the slide prior to applying the
preferred mounting medium.
 After draining the excess of clearing solvent,
a slight amount of mounting medium is
deposited on the sections.
 Some suggested first adding a minimum
quantity of medium directly to the slide to
ease positioning the thin sections.
 With the tip of a glass rod (or a pipette), a few
drops of the medium are applied in the center
of the sections and allowed to spread a
moment.
 In general 1–3 drops (or a thin line) are
sufficient.
 It is also important to assure that the medium
maintains optimal viscosity as it will normally
slowly dry in the course of the day.
 Once the mounting medium is applied, the
sections are quickly covered with a coverslip.
 If the sections curl, they should carefully be
straightened with fine tip tweezers in one
hand and the angled portion of an angled
dissecting needle in the other.
 In insufficient quantity, an imperfect seal will
be created allowing air pockets or bubbles to
form and be drawn under the coverslip as the
solvent evaporates.
 Once mounted, the excess mounting medium
should be removed to avoid later cleanup
problems.
 Great care is required when manipulating a
freshly prepared slide because the coverslip can
easily be dislodged and the tissue damaged.
 Given that mounted sections may be of uneven
thickness it may be necessary to flatten them by
placing on the cover slip one or two small lead
weights (about 25 g each) or using a weak
magnet and an iron plate.
 This procedure also helps prevent air bubbles
forming under the covers lip as the
preparation dries.
 Excessive compression should be avoided as
it may squeeze out too much of the mounting
medium favoring the formation of air bubbles
underneath the cover slip as the solvent
evaporates.
 Once mounted, the slide can be left to dry at
room temperature for weeks to months.
 The process can be hastened by placing the
slides at 40–60 °C in a drying oven or on a
slide warming plate for many hours, days, or
weeks.
 It is essential to initially store all slides in a
horizontal position until the medium is
properly hardened.
 There is nothing worse than finding out many
weeks after drying that the cover slip has
moved and the storing box is soiled with
mounting medium.
 Extreme precautions should be taken if
freshly mounted slides are to be examined
under a compound microscope as traces of
mounting medium on an objective may be
difficult to remove.
 Once slides are dried and cleaned, a proper label
should be applied to each one.
 Basic labeling should have been carried during all
procedures from the sample preparation to the
mounting and drying procedures.
 Proper labeling is crucial and it cannot be
stressed enough that thin sections from various
origins cannot be mixed up.
 At this point, a self-adhesive label may be placed
on the left side of the slide.
 Alternatively permanent ink can be used.
 The information provided on each slide label
may include a sample ID, collection date,
species name, exceptional feature in the
preparation (may be marked on the underside
of the slide with ink in case an immersion
lens should be used), as well as other
pertinent details regarding the sample and
the investigation.
 For example, if a cr oss-dated tree-ring
sequence is mounted, the date of the first
and last complete rings will be indicated.
 Once properly labeled, the slide will be stored
usually in specific slide boxes.