PMID- 30125088
OWN - NLM
STAT- MEDLINE
DCOM- 20190412
LR  - 20190412
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 90
IP  - 19
DP  - 2018 Oct 2
TI  - Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics.
PG  - 11470-11477
LID - 10.1021/acs.analchem.8b02630 [doi]
AB  - We present a novel method for real-time monitoring and kinetic analysis of 
      fluorescence in situ hybridization (FISH). We implement the method using a 
      vertical microfluidic probe containing a microstructure designed for rapid 
      switching between probe solution and nonfluorescent imaging buffer. The FISH 
      signal is monitored in real time during the imaging buffer wash, during which 
      signal associated with unbound probes is removed. We provide a theoretical 
      description of the method as well as a demonstration of its applicability using a 
      model system of centromeric probes (Cen17). We demonstrate the applicability of 
      the method for characterization of FISH kinetics under conditions of varying 
      probe concentration, destabilizing agent (formamide) content, volume exclusion 
      agent (dextran sulfate) content, and ionic strength. We show that our method can 
      be used to investigate the effect of each of these variables and provide insight 
      into processes affecting in situ hybridization, facilitating the design of new 
      assays.
FAU - Ostromohov, Nadya
AU  - Ostromohov N
AUID- ORCID: 0000-0002-7677-3099
AD  - IBM Research-Zurich , Saumerstrasse 4 , 8803 Ruschlikon , Zurich , Switzerland.
AD  - Faculty of Mechanical Engineering , Technion-Israel Institute of Technology , 
      Haifa 3200003 , Israel.
FAU - Huber, Deborah
AU  - Huber D
AD  - IBM Research-Zurich , Saumerstrasse 4 , 8803 Ruschlikon , Zurich , Switzerland.
FAU - Bercovici, Moran
AU  - Bercovici M
AUID- ORCID: 0000-0002-7803-4791
AD  - Faculty of Mechanical Engineering , Technion-Israel Institute of Technology , 
      Haifa 3200003 , Israel.
FAU - Kaigala, Govind V
AU  - Kaigala GV
AUID- ORCID: 0000-0003-1145-7287
AD  - IBM Research-Zurich , Saumerstrasse 4 , 8803 Ruschlikon , Zurich , Switzerland.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180831
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (DNA Probes)
RN  - 0 (Formamides)
RN  - 4781T907ZS (formamide)
RN  - 9042-14-2 (Dextran Sulfate)
SB  - IM
MH  - DNA Probes/chemistry
MH  - Dextran Sulfate/chemistry
MH  - Formamides/chemistry
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Kinetics
MH  - MCF-7 Cells
MH  - Osmolar Concentration
EDAT- 2018/08/21 06:00
MHDA- 2019/04/13 06:00
CRDT- 2018/08/21 06:00
PHST- 2018/08/21 06:00 [pubmed]
PHST- 2019/04/13 06:00 [medline]
PHST- 2018/08/21 06:00 [entrez]
AID - 10.1021/acs.analchem.8b02630 [doi]
PST - ppublish
SO  - Anal Chem. 2018 Oct 2;90(19):11470-11477. doi: 10.1021/acs.analchem.8b02630. Epub 
      2018 Aug 31.