Amplification of mRNAs from Single, Fixed, TUNEL-Positive Cells

O’Dell, Dianne M.; Raghupathi, Revathi K.; Crino, Peter B.; Morrison, Barclay; Eberwine, James; McIntosh, Tracy K.

doi:10.2144/98254bm04

Cited by 24 publications

(15 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Samples containing as few as 1 -10 cells can be isolated in this manner, and subsequently used in microarray experiments (Kamme et al, 2003;Seshi et al, 2003). Alternatively, similar samples can also be obtained through manual microdissection, where single-cell isolation can be accomplished using micromanipulators and pulled glass micropipettes as scalpels (Eberwine et al, 1992;O'Dell et al, 1998). Once separated from the cell matrix, the cell body is aspirated into the micropipette and placed into a sample tube for analysis.…”

Section: Microarray Technologymentioning

confidence: 99%

Expression profiling of small cellular samples in cancer: less is more

Glanzer

Eberwine

2004

Br J Cancer

View full text Add to dashboard Cite

Section: Microarray Technologymentioning

confidence: 99%

Expression profiling of small cellular samples in cancer: less is more

Glanzer

Eberwine

2004

Br J Cancer

View full text Add to dashboard Cite

“…Furthermore, specific cell types could be further characterized by looking at the gene expression pattern of these particular cells. Hitherto, gene expression profiling of single cells has been performed using aspiration of the intracellular contents of live cells or through manual dissection of fixed cells using a needle or scalpel (Dell et al, 1998). After RNA amplification, the expression of genes has been detected using radioactive labels, usually on limited, membrane-bound arrays (Mackler et al, 1992;BrooksKayal et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Single-Cell Microarray Analysis in Hippocampus CA1: Demonstration and Validation of Cellular Heterogeneity

et al. 2003

View full text Add to dashboard Cite

Laser capture microdissection in combination with microarrays allows for the expression analysis of thousands of genes in selected cells. Here we describe single-cell gene expression profiling of CA1 neurons in the rat hippocampus using a combination of laser capture, T7 RNA amplification, and cDNA microarray analysis. Subsequent cluster analysis of the microarray data identified two different cell types: pyramidal neurons and an interneuron. Cluster analysis also revealed differences among the pyramidal neurons, indicating that even a single cell type in vivo is not a homogeneous population of cells at the gene expression level. Microarray data were confirmed by quantitative RT-PCR and in situ hybridization. We also report on the reproducibility and sensitivity of this combination of methods. Single-cell gene expression profiling offers a powerful tool to tackle the complexity of the mammalian brain.

show abstract

“…Gene expression in fixed material has been investigated with different fixatives in various tissues, including the brain (Cummings et al, 2001;Relf et al, 2002;Scheidl et al, 2002;Van Deerlin et al, 2002). Each fixation method was found to have a specific impact on the integrity and yield of mRNA (O'Dell et al, 1998;Srinivasan et al, 2002), probably introducing errors in the interpretation of expression data. Formalin fixed, paraffin embedded tissues in general gave rise to a low yield and rather poor quality of RNA, whereas ethanol fixed tissue yielded a relatively intact RNA gene expression profile (Finke et al, 1993;Foss et al, 1994;Karlsen et al, 1994;Macabeo-Ong et al, 2002;Rupp and Locker, 1988;Serth et al, 2000;Shibutani et al, 2000;Specht et al, 2001;Stanta and Schneider, 1991).…”

Section: Introductionmentioning

confidence: 99%