Henderson lab - standard sample blocking for prognostic studies (neuro pg.1)

lumbar s.c. cervical s.c.

hindlimb

brain

*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/ DRG’s used. Samples A-F (vial 1), samples G-J (vial 2). Tails segments (K) from each animal go in –200C freezer.

1 2

3

4

5

6

7

8

9

10

11

A

BC

D

E

F

L2

(H)L3

L4

L5

(I)

(J)

T2

cassette

T13

L1

(G)

Henderson lab - standard sample blocking for prognostic studies (neuro pg.2)

-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)

Initial exam: 2 samples per genotype

Paraffin prep. (1) Frozen prep. (1)

-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)

7x4 = 28 um / slide6 slides / set = 168 um / setIndicate interval between sets (if any)P

roj.

No.

– e

xp.

No.

– s

lide

No.

Ex.

RO

D

-A

#1

2

7u

m