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Making a Permanent Mounted Slide
A drop of pond water under a coverslip is a wet mount, and it has about twenty minutes to live. The water creeps out, the coverslip drifts, air pushes in from the edges, and whatever you found is gone before you can show it to anyone.
A permanent mount replaces the water with a resin that sets clear and hard, sealing the specimen between glass forever. Victorian slide-makers turned this into an industry — there are mounted diatom arrangements from the 1870s that are still perfect — and the chemistry has barely changed.
The reason it works is not just preservation. The mounting medium is chosen to have a refractive index close to glass, so the specimen stops scattering light at every boundary and detail that was lost in glare simply appears. A good mount makes a specimen LOOK better, not only last longer.
Pemula
3 hours
Instruksi
1
1
Clean the glass properly
Clean the glass properly
Wash slides and coverslips in detergent, rinse, dip in alcohol and dry with a lint-free cloth. Handle them by the edges only.
Grease makes the mountant pull away into islands as it dries, leaving a permanently ruined slide that looks fine while it is wet. A fingerprint at 400× is a landscape. This is the same lesson as cleaning a TLC plate, and it fails the same way: silently, later, and only after you have done all the other work.
Material untuk langkah ini:
Kaca objek dengan kaca penutup20 buah
Etanol mutu laboratorium, 95 %100 mlAlat yang dibutuhkan:
Sarung Tangan Nitril2
2
Fix the specimen so it stops changing
Fix the specimen so it stops changing
Soak the specimen in a fixative — formalin, or a simple alcohol-acetic acid mixture — for long enough to reach the middle of it. Thin material needs minutes, thick material needs hours.
Fixing cross-links the proteins so the tissue stops decaying and holds its shape through everything that follows. Skip it and your beautiful section turns to mush in the alcohol. Work with formalin only in good ventilation and gloves — the vapour is an irritant and a sensitiser, and an alcohol-acetic fixative is perfectly adequate for plant material.
Material untuk langkah ini:
Larutan formaldehida (formalin 37 %)50 mlEtanol mutu laboratorium, 95 %200 ml
Asam asetat (glasial)20 mlAlat yang dibutuhkan:
Stoples KacaSarung Tangan Nitril
Kacamata Pelindung Tertutup3
3
Dehydrate in steps, never in one jump
Dehydrate in steps, never in one jump
Move the specimen up a series of alcohol baths — 30, 50, 70, 90 and finally 100 per cent — giving it time in each.
Water and resin do not mix, so every trace of water must go before mounting. But dropping wet tissue straight into absolute alcohol pulls water out so fast that cells shrink and distort, and delicate structures collapse. The graded series is slow and it is the difference between a section that looks like tissue and one that looks like a raisin.
Material untuk langkah ini:
Etanol mutu laboratorium, 95 %500 mlAlat yang dibutuhkan:
Stoples Kaca
Stopwatch4
4
Mount without trapping air
Mount without trapping air
Put a single small drop of mountant on the slide, lower the coverslip from one edge at a shallow angle, and let it fall the rest of the way under its own weight.
Dropping it flat traps a sheet of bubbles that you will never get out. Lowering from one edge pushes the air ahead of the advancing liquid. Use less mountant than feels right: too much oozes out and takes weeks to harden, too little pulls air in from the edges as it shrinks. One small drop for a standard coverslip.
Material untuk langkah ini:
Medium perekat preparat mikroskop10 mlAlat yang dibutuhkan:
Pipet berskala (Mohr)5
5
Dry it flat, and label it immediately
Dry it flat, and label it immediately
Lay the slide flat, somewhere warm and dust-free, for days to weeks depending on the mountant. Write the label as soon as the coverslip is down.
Tilted slides let the coverslip creep and the specimen ends up off-centre or half out. And an unlabelled slide is worthless within a month — you will not remember what it was, where it came from or how it was stained. Species or material, source, date, fixative, stain: five fields, thirty seconds, and the slide stays useful for a century.
6
6
Why the mountant makes things visible
Why the mountant makes things visible
Memuat notebook Jupyter…
Material untuk langkah ini:
Gliserin (nabati)50 ml7
7
Ring the semi-permanent mounts
Ring the semi-permanent mounts
For a glycerol or water mount, paint a ring of clear nail varnish around the coverslip edge in two thin coats, letting the first dry.
The ring does two jobs: it stops evaporation and it glues the coverslip down so an objective cannot drag it. Two thin coats beat one thick one, which shrinks as it dries and pulls away from the glass. Victorian mounters made this decorative — coloured rings and gilt paper — and the practical version is exactly the same thing without the flourish.
Bahan
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Alat yang Diperlukan
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