Biomedical Engineering Reference
In-Depth Information
-
SSC (sodium chloride-sodium citrate buffer)
Dissolve 8.77 g NaCl (150 mMoles) and 2.88 g citric acid (15
mMoles) in 800 ml ddH
2
O. Adjust pH to 7.0 with 0.5 N sodium
hydroxide, then fill up to 1000 ml.
10
×
SSC means the tenfold concentrated buffer. If 10
×
SSC was
10 × SSC
adjusted to pH 7.0, after dilution to working concentration the
pH is different to 7.0, but this difference is negligible in most
cases.
-
TBS (Tris-bufferd physiologic saline)
Dissolve 4.25 g Tris (35 mMoles), 7.6 g NaCl (130 mMoles), 0.47 g
KCl (2.5 mMoles), and 0.2 g NaN
3
(should be substituted by
other biocides, because azide is an inhibitor for iron-containing
enzymes) in 800 ml ddH
2
O.AdjustthewantedpHbetween7.6
and 8.0 with 1 N hydrochloric acid, then fill up to 1000 ml.
TBS should not be prepared with pH below 7.4, because the
buffer capacity decreases considerably.
TBSmaybemadeastenfoldstockandstoredfrozeninaliquots.
TBS stock
Sometimes you will find the following composition for TBS:
12.11 g Tris (0.1 Mole), 2.05 g NaCl (35 mMoles), 0.75 g glycine
(10 mMoles), 0.2 g NaN
3
or Thimerosal per 1000 ml, pH 7.8 - 8.4.
-
TE-Buffer
Dissolve 1.21 g Tris (10 mMoles) and 0.372 g EDTA, disodium
salt dihydrate (1 mMole EDTA) in 900 ml ddH
2
O, adjust pH 8.0
with hydrochloric acid and fill up to 1000 ml. Aliquot the buffer,
autoclave at 121
◦
C and store at 4 - 8
◦
C.
A tenfold stock of TE buffer is possible. Check pH after dilution
TE stock
to working concentration, because the pK of Tris has a relatively
large temperature and concentration dependency.
7.4.2 Buffers and Media for Tissue and Cell Culture
and Organ Perfusion
Most of the buffers are available as pre-mixed solids or ready-to-
use solutions, but preparation of these buffers makes it possible to
modify the composition.
The amount of components is given in milligrams per liter
(mMoles/l in parenthesis). If optically active substances are indi-
cated, use the naturally occurring isomer. Because the addition of
antibiotics depends on the intended purpose, they are not listed.
Buffers containing sodium (hydrogen) carbonate are not al-
lowed to be autoclaved. These buffers have to be sterile filtered
through a 0.2-
µ
m filter.
-
BasalMediumEaglewithEarle'sBalancedSalts
(BME modified)
CaCl
2
200 (1.80), KCl 400 (5.36), MgSO
4
7H
2
O 100 (0.41), NaCl
6800 (116.44), NaHCO
3
2200 (26.16), NaH
2
PO
4
·
·
H
2
O 125 (0.80);
Glucose 1000 (5.55), phenol red 10;
