Chapter 10
Archival Cytogenetic Slides
ERICH GEBHART
Introduction
A great number of cytogenetic slides stored worldwide are available for
FISH studies by the use of a recovery technique which would allow the use
of, e.g., archived tumor material. This permits the application of various
FISH techniques - particularly interphase FISH - to reevaluate chromosomal anomalies for summary studies of the diagnosis, prognosis and
therapeutic response (Schad, Dewald 1995).
A rather uncomplicated technique will be reported which reactivates
slides made permanent by embedding in Eukitt (storage age up to 15
years). It is based on the experience obtained from examinations on archival cytogenetic slides prepared from 33 breast carcinomas, from 23 cancerous effusions, from 100 rectal tumors (Gebhart et al1996), and from 65
bone marrow samples of human leukemias.
Outline
This technique consists of :
A coverslip removal procedure; a pretreatment step for exclusion of vestiges of RNA or proteins, and a modified FISH protocol. The technique can
be applied to slides made from the bone marrow of leukemic patients as
well as from slides prepared from cell suspensions of human solid tumors
and leukemias.
Erich Gebhart, Universitat Erlangen/Niirnberg, Institut fUr Humangenetik,
Schwabachanlage 10, Erlangen, 91054, Germany (phone +49-9131-8522351;
fax +49-9131-209297; e-mail egebhart@ humgenet.uni-erlangen.de)
PROTOCOL
Archival Cytogenetic Slides
ERICH GEBHART
Introduction
A great number of cytogenetic slides stored worldwide are available for
FISH studies by the use of a recovery technique which would allow the use
of, e.g., archived tumor material. This permits the application of various
FISH techniques - particularly interphase FISH - to reevaluate chromosomal anomalies for summary studies of the diagnosis, prognosis and
therapeutic response (Schad, Dewald 1995).
A rather uncomplicated technique will be reported which reactivates
slides made permanent by embedding in Eukitt (storage age up to 15
years). It is based on the experience obtained from examinations on archival cytogenetic slides prepared from 33 breast carcinomas, from 23 cancerous effusions, from 100 rectal tumors (Gebhart et al1996), and from 65
bone marrow samples of human leukemias.
Outline
This technique consists of :
A coverslip removal procedure; a pretreatment step for exclusion of vestiges of RNA or proteins, and a modified FISH protocol. The technique can
be applied to slides made from the bone marrow of leukemic patients as
well as from slides prepared from cell suspensions of human solid tumors
and leukemias.
Erich Gebhart, Universitat Erlangen/Niirnberg, Institut fUr Humangenetik,
Schwabachanlage 10, Erlangen, 91054, Germany (phone +49-9131-8522351;
fax +49-9131-209297; e-mail egebhart@ humgenet.uni-erlangen.de)
PROTOCOL