Application of three-dimensional fluorescence in situ hybridization to human preimplantation genetic diagnosis.
OBJECTIVE
To report a simple and efficient fluorescence in situ hybridization (FISH) method for preimplantation genetic diagnosis (PGD).


METHODS
Technique and method.


METHODS
A hospital in vitro fertilization (IVF) laboratory.


METHODS
Women undergoing IVF or intracytoplasmic sperm injection (ICSI).


METHODS
None.


METHODS
The intensity and clarity of signals, technical difficulty, the percentage of successfully treated blastomeres, and blastomere integrity after FISH.


RESULTS
This paraformaldehyde fixation technique simplified the process of fixation of blastomeres for PGD without losing blastomeres during fixation. A total of 35 blastomeres derived from 10 arrested embryos or abnormally fertilized eggs (one pronucleus or three pronuclei) were used for three-dimensional (3D) FISH staining. Signals in all blastomeres were obtained successfully by this method. Approximately 0.1 microL of DNA probe was enough for the detection of the signals in each blastomere, less than the volume (1 microL) used in the conventional FISH.


CONCLUSIONS
The 3D-FISH technique for PGD is easy to learn, less damaging to blastomeres, and loses no blastomeres during fixation. It is efficient, feasible, and economic, which allows more patients to benefit from this technique.
Yan
Li-Ying
LY
Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China.
Qiao
Jie
J
Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China. Electronic address: jie.qiao@263.net.
Chen
Yuan
Y
Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China.
Huang
Jin
J
Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China.
Liu
Ping
P
Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China.
Sun
Qing-Yuan
QY
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.
D001755:Blastocyst / Q000166:cytology; D001757:Blastomeres / Q000166:cytology; D003362:Cost-Benefit Analysis; D004526:Efficiency; D005240:Feasibility Studies; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000191:economics / Q000379:methods*; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000191:economics / Q000379:methods*