Biology Reference
In-Depth Information
Table 1
Phosphate buffer composition (mixing ratios) to gain required pH
x
y
pH
x
y
pH
93.5
6.5
5.7
45.0
55.0
6.9
92.0
8.0
5.8
39.0
61.0
7.0
90.0
10.0
5.9
33.0
67.0
7.1
87.7
12.3
6.0
28.0
72.0
7.2
85.0
15.0
6.l
23.0
77.0
7.3
81.5
18.5
6.2
19.0
81.0
7.4
77.5
22.5
6.3
16.0
84.0
7.5
73.5
26.5
6.4
13.0
87.0
7.6
68.5
31.5
6.5
10.5
89.5
7.7
62.5
37.5
6.6
8.5
91.5
7.8
56.5
43.5
6.7
7.0
93.0
7.9
51.0
49.0
6.8
5.3
94.7
8.0
Table 2
Composition of individual steps of ethanol-butanol dehydration series
Step no.
Distilled water (%)
Ethanol (%)
Butanol (%)
1
70
20
10
2
60
25
15
3
45
30
25
4
30
30
40
5
20
25
55
6
10
20
70
7
-
15
85
8
-
-
100
2.2 Whole-Mount
Preparation
NaI-based clearing solution for whole mounts: Dissolve 0.04 g of
Na
2
S
2
O
3
in 20 ml of 65 % (aq. v/v) glycerol. Add and dissolve 17 g
of NaI, 2 % (v/v) of DMSO to fi nal solution. The fi nal solution
should be clear and colorless with refractive index close to 1.5.
2.3 Dehydration,
Paraffi n Infi ltration,
and Embedding
1. Ethanol-butanol dehydration series: Composition of individ-
ual steps is specifi ed in Table
2
.
2. Anhydrous ethanol and butanol: To effi ciently remove water
from the standard stock butanol or 96 % ethanol (
see
ref. [
40
]),
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