Advanced 3D Analysis and Optimization of Single-Molecule FISH in Drosophila Muscle.Single-molecule fluorescence in situ hybridization (smFISH) provides direct access to the spatial relationship between nucleic acids and specific subcellular locations.  Although smFISH is well established in cell culture or thin sections, the utility of smFISH is hindered in thick tissue sections due to the poor probe penetration of fixed tissue, the inaccessibility of target mRNAs for probe hybridization, high background fluorescence, spherical aberration along the optical axis, and the lack of methods for image segmentation of organelles.  Studying mRNA localization in 50 μm thick Drosophila larval muscle sections, these obstacles are overcome using sample-specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple-organelle segmentation. 
smFISH
Advanced 3D Analysis and Optimization of Single-Molecule FISH in Drosophila Muscle.Single-molecule fluorescence in situ hybridization (smFISH) provides direct access to the spatial relationship between nucleic acids and specific subcellular locations.  Although smFISH is well established in cell culture or thin sections, the utility of smFISH is hindered in thick tissue sections due to the poor probe penetration of fixed tissue, the inaccessibility of target mRNAs for probe hybridization, high background fluorescence, spherical aberration along the optical axis, and the lack of methods for image segmentation of organelles.  Studying mRNA localization in 50 μm thick Drosophila larval muscle sections, these obstacles are overcome using sample-specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple-organelle segmentation. 
smFISH
Advanced 3D Analysis and Optimization of Single-Molecule FISH in Drosophila Muscle.Single-molecule fluorescence in situ hybridization (smFISH) provides direct access to the spatial relationship between nucleic acids and specific subcellular locations.  Although smFISH is well established in cell culture or thin sections, the utility of smFISH is hindered in thick tissue sections due to the poor probe penetration of fixed tissue, the inaccessibility of target mRNAs for probe hybridization, high background fluorescence, spherical aberration along the optical axis, and the lack of methods for image segmentation of organelles.  Studying mRNA localization in 50 μm thick Drosophila larval muscle sections, these obstacles are overcome using sample-specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple-organelle segmentation. 
smFISH
Advanced 3D Analysis and Optimization of Single-Molecule FISH in Drosophila Muscle.Single-molecule fluorescence in situ hybridization (smFISH) provides direct access to the spatial relationship between nucleic acids and specific subcellular locations.  Although smFISH is well established in cell culture or thin sections, the utility of smFISH is hindered in thick tissue sections due to the poor probe penetration of fixed tissue, the inaccessibility of target mRNAs for probe hybridization, high background fluorescence, spherical aberration along the optical axis, and the lack of methods for image segmentation of organelles.  Studying mRNA localization in 50 μm thick Drosophila larval muscle sections, these obstacles are overcome using sample-specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple-organelle segmentation. 
smFISH
Studying mRNA localization in 50 μm thick Drosophila larval muscle sections, these obstacles are overcome using sample-specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple-organelle segmentation. 
maximum likelihood testing
Advanced 3D Analysis and Optimization of Single-Molecule FISH in Drosophila Muscle.Single-molecule fluorescence in situ hybridization (smFISH) provides direct access to the spatial relationship between nucleic acids and specific subcellular locations.  Although smFISH is well established in cell culture or thin sections, the utility of smFISH is hindered in thick tissue sections due to the poor probe penetration of fixed tissue, the inaccessibility of target mRNAs for probe hybridization, high background fluorescence, spherical aberration along the optical axis, and the lack of methods for image segmentation of organelles.  Studying mRNA localization in 50 μm thick Drosophila larval muscle sections, these obstacles are overcome using sample-specific optimization of smFISH, particle identification based on maximum likelihood testing, and 3D multiple-organelle segmentation. 
smFISH