Single cell gene expression profiling

Multiplexed expression fluorescence in situ hybridization: Application to the analysis of cultured cells

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Most current methods of measuring gene expression rely on averaging many cellular responses or artificial amplification steps to reach a detectable threshold of signal. Columns are fashioned by removing the cotton from the top of one of these pipettes and using a portion of it to plug up the tip. To ease data processing and avoid manual manipulations that introduce bias, one requires coding the filtering and data analysis software in the JAVA programming language using the Java development kit and advanced imaging library from Sun Microsystems. Preparation of cell samples involves sterilizing a box of coverslips by boiling in 0.1 N HCl for 20 min. Rinsing and washing the coverslips in DDW several times. One needs to test hybridization before color coding and multiple transcript detection, which is accomplished using two bright dyes to show transcription sites. After this, assign each gene an arbitrary color code using combinations of dyes and test singly.

Original languageEnglish (US)
Title of host publicationCell Biology, Four-Volume Set
PublisherElsevier Inc.
Pages121-126
Number of pages6
Volume4
ISBN (Print)9780121647308
DOIs
StatePublished - 2006

Fingerprint

Gene Expression Profiling
Fluorescence In Situ Hybridization
Gene expression
Cultured Cells
Coloring Agents
Color codes
Color
Fluorescence
Cells
Programming Languages
Microsystems
Solar System
Transcription
Washing
Sun
Computer programming languages
Boiling liquids
Cotton
Libraries
Amplification

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Single cell gene expression profiling : Multiplexed expression fluorescence in situ hybridization: Application to the analysis of cultured cells. / Levsky, Jeffrey M.; Braut, Steven A.; Singer, Robert H.

Cell Biology, Four-Volume Set. Vol. 4 Elsevier Inc., 2006. p. 121-126.

Research output: Chapter in Book/Report/Conference proceedingChapter

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