Submission date

Name of primary investigator

Tissue type, Case number, special characteristics

 
1.     Tissue Processing

Primary fixation

(1 hour)

Fix tissue in 2.5% glutaraldehyde, 2.0% formaldehyde in 0.1M Na cacodylate buffer, pH 7.4, for 2 h.

Wash

(2 hours)

Wash at least 3X in 0.1M Na cacodylate, pH 7, 20 min.

Post-fixation

(1 hour)

1% OsO4 with 0.5% K4Ferro(CN)6 in 0.1M Na cacodylate buffer for 1 h at RT followed by 1 wash in buffer then water.

Dehydration

(1.5 hours)

50, 75, 80, 90, 95% ethanol, 10 min each, 100% ethanol, 5 min X 3

Propylene oxide X 2

Infiltration

(3 hours then overnight)

1:2 Epoxy:PO 2 h (Epoxy = Epon Araldite)

1:1 Epoxy:PO 2 h

3:1 Epoxy:PO Overnight

100% Epoxy 2 changes then ON on rocker.

Polym erization

(2d)

Transfer to fresh epoxy beds in BEEM caps, heat cure at 65C for 2 days.

 

  1. Ultramicrotomy, post stain thin sections

 

  1. TEM exam, photography

 

4.     Develop films, Scan and send images to PI

 

Blocks list:

Description

Block #

Comment

 

05RPxxx

 

 

Results Summary: