Chapter 7
In Situ Hybridization for RNA:
Radioactive DNA Probe
T AKASHI SAW AI AND MIW A UZUKI
Introduction
In situ hybridization (ISH) was first described in 1969 by Pardue
and Gall. Tritium labeled nucleic acid was first used and detected
via exposure to a nuclear track emulsion. Though ISH has been
improved over the years, it still takes a long time to obtain fine
and precise results especially when using tritium labeled probes
as compared to hours of those non-radioacitve probes. The other
potential labels such as 125 1 and 32p for ISH were recognized in
the early 1970s. The limitations of radioactive isotopic detection
for ISH were recognized from an early stage. Today in vitro biochemical labeling of purified nucleic acids is universally employed and in the future it seems that chemically synthesized
oligonucleotides will be used.
The combination of radioisotopes and detection via a contact
emulsion has been successfully used for many years. We have
applied wide biochemical research such as in the field of developmental biology and pathology. Researchers need to be
aware of the underlying principles of detection to select the
best combination of radiolabeling and assay conditions to
suit the particular experiment. Although ISH with the radioactive method is rather old, it still has the fundamental principles of
understanding ISH. Our hybridization procedures are basically
the same as those described by Hayashi et al. 1986, which is the
modified Hafen's method (Hafen et al. 1983).
Correspondence to Takashi Sawai, Iwate Medical University, Dept. of
Pathology, 19-1 Uchimaru, Morioka-City, 020-8505, Japan (phone +8119-651-5111 ext. 3510; fax +81-19-651-9246; e-mail tsawai@iwatemed.ac.jp)
In Situ Hybridization for RNA:
Radioactive DNA Probe
T AKASHI SAW AI AND MIW A UZUKI
Introduction
In situ hybridization (ISH) was first described in 1969 by Pardue
and Gall. Tritium labeled nucleic acid was first used and detected
via exposure to a nuclear track emulsion. Though ISH has been
improved over the years, it still takes a long time to obtain fine
and precise results especially when using tritium labeled probes
as compared to hours of those non-radioacitve probes. The other
potential labels such as 125 1 and 32p for ISH were recognized in
the early 1970s. The limitations of radioactive isotopic detection
for ISH were recognized from an early stage. Today in vitro biochemical labeling of purified nucleic acids is universally employed and in the future it seems that chemically synthesized
oligonucleotides will be used.
The combination of radioisotopes and detection via a contact
emulsion has been successfully used for many years. We have
applied wide biochemical research such as in the field of developmental biology and pathology. Researchers need to be
aware of the underlying principles of detection to select the
best combination of radiolabeling and assay conditions to
suit the particular experiment. Although ISH with the radioactive method is rather old, it still has the fundamental principles of
understanding ISH. Our hybridization procedures are basically
the same as those described by Hayashi et al. 1986, which is the
modified Hafen's method (Hafen et al. 1983).
Correspondence to Takashi Sawai, Iwate Medical University, Dept. of
Pathology, 19-1 Uchimaru, Morioka-City, 020-8505, Japan (phone +8119-651-5111 ext. 3510; fax +81-19-651-9246; e-mail tsawai@iwatemed.ac.jp)