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Center for Cancer Research National Cancer Institute
Tissue Array Research Program (TARP)

Protocols

The following suggested protocols may be used when using the multi-tumor tissue microarray slides.

H&E Protocol For Array Slides On Tape Section Slides

The following suggested protocol was provided by
David E. Kleiner M.D., Ph.D and Stephen M. Hewitt, M.D.,Ph.D.

  • Staining the Instrumedic's acrylic coated slides used for tape sectioning.
  • It may be used for general H&E's as well.
  • This stain is programmed on our Leica Autostainer XL.
  • Slides cut onto regular slides may be stained per routine.

Step

Reagent

Time

1

Xylene

2 minutes

2

Xylene

2 minutes

3

Xylene

2 minutes

4

100% Ethanol

1 minute

5

100% Ethanol

1 minute

6

95% Ethanol

1 minute

7

95% Ethanol

1 minute

8

Water

2 minutes

9

Hematoxylin1

3 minutes

10

Water

2 minutes

11

Bluing Agent2

30 seconds

12

Water

30 seconds

13

Eosin Aqueous3

3 minutes

14

Water

30 seconds

15

Oven 65°

2 minutes

16

100% Ethanol

1 minute

17

100% Ethanol

1 minute

18

Xylene

1 minute

19

Xylene

1 minute

20

Xylene

1 minute

  1. We use Lerner-2 double strength hematoxylin, 0.36% hematoxylin.
  2. We use a LiCo3 0.044%, methanol 50%, bluing agent.
  3. It is essential aqueous eosin is used, or the acrylic adhesive on the slides will be stained.
    We use Shandon Eosin Y Aqueous, 0.5% Eosin Y.

For more technical information only (no ordering, shipping, or billing inquiries, please), contact the TARP Lab at 301-435-2951, or hewitts@mail.nih.gov.

Added by Cline, Ave (NIH/NCI) [E] , last edited by Cline, Ave (NIH/NCI) [E] on Jul 08, 2011 09:28