Penempelan (affixing)

 Hasil sayatan blok pada tahapan sectioning kemudian

ditempelkan pada permukaan slide
 Penempelan membutuhkan reagen tertentu agar dapat
melekat erat dan tidak lepas selama proses pewarnaan
 Reagen yang dapat digunakan:
 Albumin mayer (merupakan campuran antara albumin dan gliserol)
 Lem pabrikan (merupakan reagen yang lebih praktis)
 Haupt’s affixative
 Celloidin
There are several commonly used affixatives

1. Haupt’s affixative contains :

100 ccs (cubic centimeter) distilled water,
1gm gelatin,
2 gm phenol crystals,
15 cc glycerine.
To make it,

1. Dssolve 1 g. plain Knox gelatin (use the type that comes

four envelopes to the package) or other pure, finely
divided gelatin in 100 cc distilled water at a temperature of
30°C.)
2. When completely dissolved, add 2 g. phenol crystals and
15 cc glycerin:
3. Stir well, then filter.
How to use :
1. A 3 to 4% aqueous solution of formalin is used for floating sections.
2. Place a drop of the adhesive on the clean slide, smear evenly so as
to leave a barely perceptible layer,
3. then immediately add the formalin solution by means of a pipette.
4. Place the sections on the formalin solution,
5.put the slide on a warming table (at a temperature of between 40 and
43°C. for; paraffin with a melting point around 58 to 60°C.) until the
sections have straightened out.
6. Remove the slide from the warming table, and set aside until the
waterbecomes cool;
7. Drain off excess water, arrange the sections as desired, and set the
slide aside to dry.

By putting mounted but still wet slides in a drying oven together with a watch glass or two of full
strength formalin, the fumes of the formalinwill further assist in coagulating the gelatin of the adhesive.
-
In botanical microtechnique,
adhesives are fluids or substances used for affixing paraffin sections
to slides.

Haupt's Adhesive by far is the best fluid yet devised for the
affixing of sections to the slide

may also be used for affixing unicellular specimens

and many colonial algae.
2. Celloidin.
Thick woody sections and serial sections of certain marine algae which
are retained on the slide with the utmost difficulty may be coated with a
1 to 2% solution of celloidin (in equal parts of absolute alcohol and
ether) after first having been affixed with Haupt's adhesive and
thoroughly dried.

Remove the paraffin with carbol-xylol (10% phenol in xylol) and the latter
with 95% ethyl alcohol.

Under ordinary-circumstances the interpolation of a 1 % solution of

celloidin in equal' parts of absolute alcohol and ether in the staining
schedule between the absolute alcohol-xylol and 95% alcohol stages will
make the majority of sections adhere firmly to the slides.
2. Ullrich's Adhesive.

100 cc of distilled water

1 cc of standard water-glass solution
1 cc of concentrated ammonia

Dry thoroughly in the air after stretching the sections

with the aid'of heat.
Dissolve the paraffin with xylol as usual
Bring down the series of alcohols to 70%.
(to this percentage add a trace of hydrochloric acid.)
 Mayer's Adhesive is the older standard
adhesive.
3. Mayer’s affixative is consist of :
5 cc egg albumen,
50 cc glycerine,
1 gm sodium salicylate.

1. To the white of a fresh egg add about an equal quantity of glycerin and
1 g of sodium salicylate or a crushed crystal of thymol.
2. Shake well,
3. filter through sterile cotton or two or three thicknesses of sterile
cheese-cloth.
It can be kept for as long as 6 months, solutions more than a month old
lose their adhesive quality.
Pada mikrotehnik tumbuhan

Mayer's adhesive possesses less holding quality than

Haupt's

and also has the annoying property of absorbing coal-

tar dyes.
4. If eggs are cheap and easily available, one may
use filtered white of egg alone.
The general steps of affixing paraffin sections can be concluded as

1. Clean the required slides,

2. Mark the cleaned slides,
3. Drop affixative on each slide,
4. Put on another slide,
5. Spread the affixative,
continued ...

6. Drop floating medium,

7. Divide the paraffin into required length,
8. Transfer the sections,
9. Add more floating medium if incomplete floating occurs,
10. Rise the temperature,
11. Remove slides and redundant floating medium,
12. drying the section
Tahapan affixing

 Penempelan sayatan paraffin

 Melapisi slide dengan albumin mayer
 Meletakkan slide diatas meja lalu permukaannya ditutup dengan air
 Hasil sayatan dipotong lalu dipindahkan ke atas permukaan air diatas
slide menggunakan scalpel atau kuas yang telah dibasahi
 Slide diletakkan pada hotplate dan biarkan pita sayatan
mengembang dan merata
 Buang kelebihan air lalu keringkan hingga seluruh molekul air
menguap.
 Hal yang perlu diperhatikan selama proses
penempelan
 Gunakan slide bersih
 Lapisi slide dengan albumin mayer setipis mungkin
 Jangan menyentuh permukaan slide yang telah dilapisi albumin
mayer
 Buang seluruh albumin pada permukaan slide yang tidak ditempat
dengan tissue
 Permukaan pita sayatan yang mengkilap menghadap permukaan
slide
 Jaga agar paraffin jangan sampai mencair
 Slide yang telah ditempeli dapat disimpan dalam oven hangat
selama tiga hari
NEXT STEP

Processing paraffin sections include

1. Deparaffination,
2. Removing the deparaffing solution,
3. Hydration,
4. Staining,
5. Dehydration,
6. Dealcoholisation and clearing,
7. Mounting the cover slide