Single Molecule FISH of Lgr5 mRNA

The FISH procedure was performed according to previously published methods [47 (link)]. 7 μm cryo-sections were cut for hybridizations. Lgr5 mRNA Stellaris probe (mouse: NM_010195) was designed by Probe Designer at www.singlemoleculefish.com and synthesized by Biosearch Technologies. The FISH probe set consists of 96 TMR fluorophore labelled oligonucleotides. DAPI nuclear dye was included during the final wash. Images were captured at 405nm and 568nm using 100x objective len by an Olympus FV1000 upright confocal microscope.

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Goh A.M., Xue Y., Leushacke M., Li L., Wong J.S., Chiam P.C., Rahmat S.A., Mann M.B., Mann K.M., Barker N., Lozano G., Terzian T, & Lane D.P. (2015). Mutant p53 accumulates in cycling and proliferating cells in the normal tissues of p53 R172H mutant mice. Oncotarget, 6(20), 17968-17980.

Publication 2015

FV1000 upright confocal microscope

Confocal microscope Dapi Fish Hybridizations Mouse Mrna Oligonucleotides

Corresponding Organization :

Other organizations : Agency for Science, Technology and Research, A*STAR Graduate Academy, Institute of Medical Biology, Houston Methodist, Institute of Molecular and Cell Biology, The University of Texas MD Anderson Cancer Center

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Variable analysis

independent variables

FISH procedure performed according to previously published methods [47]

dependent variables

Lgr5 mRNA expression

control variables

7 μm cryo-sections used for hybridizations
Lgr5 mRNA Stellaris probe (mouse: NM_010195) designed by Probe Designer at www.singlemoleculefish.com and synthesized by Biosearch Technologies
FISH probe set consists of 96 TMR fluorophore labelled oligonucleotides
DAPI nuclear dye included during the final wash
Images captured at 405nm and 568nm using 100x objective lens by an Olympus FV1000 upright confocal microscope

controls

Positive control: Not specified
Negative control: Not specified

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