Assessment Run 5 2001 

CD3

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The slides to be stained for CD3 contained four T-cell lymphomas and one B-cell lymphoma in a spleen 'matrix'.

45 laboratories submitted the staining. Of these, 35 used Dako's pAb, 8 used mAb PS1, 1 used mAb F7.2.38, and 1 used mAb UCHL-1.

At the assessment optimal staining was achieved in 10, acceptable in 21, borderline in 12 and poor staining in 2 of the laboratories. Optimal results were in one or more cases achieved with all of the Abs Dako's pAb, PS1 and UCHL-1.

Mandatory for an optimal staining was an efficient HIER and appropriate dilution of the primary antibody.

The main reasons for a borderline or poor staining was insufficient HIER (too short efficient heating time and/or inappropriate pH) and a too dilute primary Ab (in one too concentrated Ab) in relation to the overall sensitivity of the protocol.

 

Fig. 1A. Optimal staining of a T-cell lymphoma, using mAb PS1 (good protocol 1). All T-cells are strongly stained, while residual B-cells (upper left corner) are unstained. Exactly the same staining result were obtained using Dako's pAb (good protocol 2).

 

Good protocol 1 Good protocol 2 

Fig. 1B (same field as in Fig. 1A, same Ab). Acceptable staining of a T-cell lymphoma. The reaction can be interpreted, but the signal is weak.
Fig 1C (same field as in Fig. 1A, same Ab). Insufficient staining. The T-cells is very faintly stained, the nuclear counterstaining predominate. Fig. 2 Staining of spleen. Insufficient staining using DAKO's polyclonal Ab. Due to high concentration of the Ab, a considerable cross reaction with B-cells is seen.

MV/SN

Last update: 01-04-07