Henderson lab standard sample blocking for prognostic studies neuro pg'1

1
Henderson lab - standard sample blocking for
prognostic studies (neuro pg.1)
lumbar s.c.
cervical s.c.
brain
cassette
1
2
(I)
6
F
A
4
9
(G)
T2
T13
B
C
3
D
L1
5
L2
E
7
Motor tests performed prior dissections.
Landmarks and positions of standard dissection
points are indicated for each segment. Cuts
(numbers) are indicated in the order performed.
Tissue segments (letters) are indicated for
database registration. Mice are perfused (4 PFA)
and all tissue segments postfixed in 4 PFA at
40C O/N. Spinal segments are postfixed in the
spinal column and removed the following day.
Yellow lines indicates which face of a tissue
segment should be placed down in the cassette for
sectioning. Circles indicate sites of VR/ DRGs
used. Samples A-F (vial 1), samples G-J (vial 2).
Tails segments (K) from each animal go in 200C
freezer.
10
(H)
L3
hindlimb
(J)
L4
L5
8
11
2
Henderson lab - standard sample blocking for
prognostic studies (neuro pg.2)
Initial exam 2 samples per genotype
Paraffin prep. (1)
Frozen prep. (1)

• Postfix 4 PFA 40C O/N
• Embedding (A-F G)
• 7 um sections taken in sets of 6
• (Th / DAPI, CR, LFA, exp. 1, 2, 3)

• Postfix 4 PFA 40C O/N
• 30 sucrose 16-24 hrs.
• Embedding in OCT (A-F G)
• Section (30 um-floating, 10 um-thaw mount)

Proj. No. exp. No. slide No.
7x4 28 um / slide 6 slides / set 168 um /
set Indicate interval between sets (if any)
Ex. ROD -A 12 7um