Konference: 2011 7. Sympozium a workshop molekulární patologie a histo-cyto-chemie
Číslo abstraktu: 008p
Autoři: A. Lodererova; MUDr. Eva Honsová, Ph.D.; L. Voska; V. Gabris; J. Maluskova
Combining two different scientific disciplines –
morphology and immunochemistry – immunohistochemistry has developed
as an important instrument in research and clinical pathology.
However, in some cases there is a need for knowledge about the
relative localization of targets, which can only be obtained by
vizualizing all relevant targets in one slide. Multiple staining
can be defined as the detection of two or more targets on one
slide. This technique enables the depiction of two or more
targets/antigens in connection with other morphological features.
We use the Benchmark ULTRA system in our lab, which is the next
generation in IHC/ISH staining instrumentation. For the double
staining method we use sequential staining. The most important step
is to optimize the pretreatment step (time, buffer or protease) for
both antigens. For the first detection step we selected ultraView
Universal DAB system (brown) and for the second one ultraView
Universal Alkaline Phosphatase Detection Kit (red).
The following combinations of antigens were
determined as helpful: Ki 67 + CK 7; Ki 67 + CD 31; CD 34 + CK 7;
CD 31 + p 53; D2- 40 + AE 1/3; CD 3 + CK 7; kappa + lambda light
Conclusion: Detection of more
targets/antigens in one slide has good results. Moreover, this
method spares the tissue, which is very important in cases with
small biopsy samples.
Datum přednesení příspěvku: 29. 4. 2011