PMID- 22541126
OWN - NLM
STAT- MEDLINE
DCOM- 20121220
LR  - 20161018
IS  - 1009-2137 (Print)
IS  - 1009-2137 (Linking)
VI  - 20
IP  - 2
DP  - 2012 Apr
TI  - [An improved protocol of preparing bone marrow cells for fluorescence in situ 
      hybridization].
PG  - 496-9
AB  - This study was aimed to establish a smear protocol for preparing bone marrow 
      cells and investigate its effect on fluorescence in situ hybridization (FISH) 
      signal. Probe DNA (C-myc, MDM2, STK6) was labeled with Spectrum Green, 
      PromoFluor-555 and PromoFluor-415 by nick translation. Five bone marrow samples 
      were tested by two methods separately. Traditional method: after removing the 
      erythrocytes by hypoosmotic solution, the bone marrow cells were fixed in 
      methanol/acetic acid (3:1). Improved method: erythrocytes were removed using 
      density gradient centrifugation and fixed in methanol. The samples were then 
      fixed again in 2 formaldehyde for 5 min. The FISH signal was assessed by 
      comparing the relative signal intensity of each fluorophore with the 
      autofluorescence background. The results indicated that improved method greatly 
      increased the ratio of fluorescence signal intensity in the Spectrum Green, 
      PromoFluor-555 and PromoFluor-415 channel (traditional method: 4.3 +/- 0.19, 3.52 +/- 
      0.04, 3.07 +/- 0.08; improved method: 9.89 +/- 0.41, 7.55 +/- 0.5, 5.67 +/- 0.18, n = 5, 
      P < 0.01) respectively. The signal intensity increased 2.32, 2.14 and 1.85-fold 
      in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel respectively. In 
      addition, the improved method decreased the split signals [traditional method: 
      (15.8 +/- 1.74), (20.42 +/- 2.88), (23.2 +/- 3.02); improved method: (8.6 +/- 1.2), 
      (12.28 +/- 1.33), (12.6 +/- 2.56), n = 5, P < 0.05]. It is concluded that the 
      improved optimal procedure which facilitates FISH intensity on bone marrow cells 
      is developed, showing potential for wide application in the diagnosis of 
      hematologic diseases.
FAU - Hu, Lin-Ping
AU  - Hu LP
AD  - Chinese Academy of Medical Sciences, Tianjin, China.
FAU - Ge, Jing
AU  - Ge J
FAU - Zhang, Li-Yan
AU  - Zhang LY
FAU - Xu, Jing
AU  - Xu J
FAU - Yuan, Wei-Ping
AU  - Yuan WP
FAU - Cheng, Tao
AU  - Cheng T
FAU - Zhang, Lei
AU  - Zhang L
LA  - chi
PT  - English Abstract
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - China
TA  - Zhongguo Shi Yan Xue Ye Xue Za Zhi
JT  - Zhongguo shi yan xue ye xue za zhi
JID - 101084424
SB  - IM
MH  - Bone Marrow Cells/*cytology
MH  - *Histocytological Preparation Techniques
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
EDAT- 2012/05/01 06:00
MHDA- 2012/12/21 06:00
CRDT- 2012/05/01 06:00
PHST- 2012/05/01 06:00 [entrez]
PHST- 2012/05/01 06:00 [pubmed]
PHST- 2012/12/21 06:00 [medline]
AID - 1009-2137(2012)02-0496-04 [pii]
PST - ppublish
SO  - Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2012 Apr;20(2):496-9.