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Chemicals used for
preservation of carcasses and samples
First
unrevised draft. Help for improvement would be appreciated
For
fixing of tissues for histology
For histological studies, buffered
neutral formalin is a good general purpose fixative. For certain histological
procedures, however, other fixatives may be better because they penetrate
tissues more rapidly and may render tissues more easily stained by certain
histological dyes. Tissues can be left in some fixatives (e.g., buffered
neutralized formalin) for several months; with other fixatives (e.g., Bouin's),
tissues must be transferred to alcohol immediately after fixing. Three
of the more widely used fixatives are described here. For more information
on fixatives and histological methods, Nagorsen and Peterson recommend
Luna (1968).
Formalin
(= formaldehyde solution):
Commercially available formalin solution
is usually a 37 % or 40 % (weight / volume) solution. It may be best to
take this full strength solution to the field to reduce volume and to dilute
it before use; in this case the full strength solution is treated as 100
%, which means that for production of a 10 % solution one part of 40 %
formalin solution must be mixed with 9 parts water. Formalin solution is
usually acidic (pH 3 to 4.6) and tends to decalcify teeth and to excessively
harden tissue. Therefore, it is best to buffer it to pH 7.0 (neutral) (Nagorsen,
Peterson 1980).
10 % buffered formalin (from
Nagorsen, Peterson 1980, Munson 2000)
For 1 litre:
100 ml formalin
(38-40 % formaldehyde)
900 ml distilled
water
Buffer added:
Buffer
mixture for precise buffering to neutrality (pH 7.0) (Nagorsen, Peterson
1980):
4 g of acid or monobasic sodium phosphate monohydrate (NAH2PO4H2O)
6.5 g of dibasic sodium phosphate anhydrate (NA2HPO4)
This dry salt mixture can be prepared in advance and carried into the field
in plastic bags.
Another
possible buffer mixture (Munson 2000):
4.5 gm sodium phosphate (monobasic)
3.6 g sodium hydroxide
Mixture
in cases when such buffer is not available:
4 g sodium chloride = table salt (Munson 2000)
or
a quarter of a teaspoon of borax powder or household ammonia (Nagorsen,
Peterson 1980) per litre
Bouin's
solution (from Nagorsen, Peterson 1980)
Picric acid, saturated
aqueous solution 750 ml
(if not stored in an aqueous solution, picric acid is highly volatile)
37-40% formalin
250 ml
Glacial acetic
acid 50 ml
Alcohol-formalin-acetic
acid solution (AFA) (from Nagorsen, Peterson 1980)
37-40% formalin
10 ml
Alcohol, 80% 90
ml
Glacial acetic
acid 5 ml
Formalin-sodium
acetate solution (from Nagorsen, Peterson 1980)
37-40% formalin
100 ml
Sodium acetate
20 g
Tap water 900
ml
Chemicals
for other purposes
Sterile
buffered glycerin (50%) for transporting tissues for culture when refrigeration
is not available (from Munson, 2000):
Buffer composed
of:
A: 21 g citric acid mixed in 1000 distilled water
B: 28.4 g anhydrous sodium phosphate in 1000 distilled water
Buffer: mixture of 9.15 ml of A and 90.85 ml of B.
100 ml of buffer mixed with 100 ml
of glycerin.
To take the mixture into the field,
it can then be sterilised in small tubes.
"Easy
Blood" for transporting DNA from blood cells for genetic studies when
refrigeration is not available and for preserving refrigerated or frozen
DNA for longer periods of time (from Munson 2002):
1.2 g Tris HCI
3.7 g Na2
EDTA
2 g sodium dodecyl
sulfate (SDS)
Water added to
100 ml
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In:
Loris and potto conservation database: field methods
http://www.species.net/primates/loris. |
Last
amendment: 7 November 2002
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