PMID- 23331574
OWN - NLM
STAT- MEDLINE
DCOM- 20150519
LR  - 20191210
IS  - 1469-8730 (Electronic)
IS  - 0967-1994 (Linking)
VI  - 22
IP  - 3
DP  - 2014 Aug
TI  - Sample preparations are important for fluorescence in situ hybridization of cells 
      biopsied from preimplantation embryos.
PG  - 300-4
LID - 10.1017/S0967199412000639 [doi]
AB  - Fluorescence in situ hybridization (FISH) is a cytogenetic technology used to 
      detect chromosomal abnormalities in preimplantation human embryos. However, its 
      efficiency is not stable due to improper sample preparation. The present study 
      was designed to modify the current sample preparation technique and then to 
      evaluate its efficiency in human preimplantation genetic diagnosis (PGD). Day 3 
      cleavage embryos as well as day 5 and 6 blastocysts were biopsied by mechanical 
      aspiration method. In the present study, two methods were used for sample 
      preparation of the biopsied cells. Method I was the traditional method, in which 
      each blastomere was placed in a hypotonic solution for 5 min and then fixed on 
      glass slides. The slides were kept at room temperature before the FISH 
      procedures. Method II was a modified method, in which all blastomeres were placed 
      individually in hypotonic solution drops covered by oil for at least 5 min and 
      then fixed on slides with 0.1% Tween/HCl. After fixation, the slides were kept at 
      -20 degrees C for at least 30 min before the FISH procedures. The two methods were 
      compared in terms of time consumption and proportions of blastomeres with FISH 
      signals. In total, 329 blastomeres from day 3 embryos were fixed by Method I with 
      an average fixation time of 8-10 min for each blastomere. By contrast, with 
      Method II, 362 blastomeres were fixed and the average time was 3-4 min for each 
      blastomere. After FISH, more nuclei had signals with Method II (97.2%) than with 
      Method I (86.9%). All cells that were biopsied from blastocysts and prepared with 
      Method II had FISH signals. However, Method I was not suitable for the fixation 
      of multiple cells biopsied from blastocysts as cells were not traceable during 
      the fixation. The present study indicates that proper sample preparation is 
      critical for obtaining FISH signals in cells biopsied from preimplantation human 
      embryos; hence these modifications can increase the efficiency of human PGD.
FAU - Li, Lifei
AU  - Li L
AD  - The Reproductive Medicine Research Center of the 1Hospital of Lanzhou 
      University,Gansu Province 730000,China.
FAU - Zhang, Xuehong
AU  - Zhang X
AD  - The Reproductive Medicine Research Center of the 1Hospital of Lanzhou 
      University,Gansu Province 730000,China.
FAU - Wang, Weihua
AU  - Wang W
AD  - Houston Fertility Institute,2500 Fondren Rd.,Suite 350,Houston,Texas,USA.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20130118
PL  - England
TA  - Zygote
JT  - Zygote (Cambridge, England)
JID - 9309124
SB  - IM
MH  - Biopsy
MH  - Blastocyst/*cytology
MH  - Blastomeres/cytology
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Preimplantation Diagnosis/*methods
EDAT- 2013/01/22 06:00
MHDA- 2015/05/20 06:00
CRDT- 2013/01/22 06:00
PHST- 2013/01/22 06:00 [entrez]
PHST- 2013/01/22 06:00 [pubmed]
PHST- 2015/05/20 06:00 [medline]
AID - S0967199412000639 [pii]
AID - 10.1017/S0967199412000639 [doi]
PST - ppublish
SO  - Zygote. 2014 Aug;22(3):300-4. doi: 10.1017/S0967199412000639. Epub 2013 Jan 18.