Laser-assisted microdissection: applications in molecular pathology

Sirivatanauksorn, Yongyut; Drury, Rosybel; Crnogorac‐Jurčević, Tatjana; Sirivatanauksorn, Vorapan; Lemoine, Nicholas R.

doi:10.1002/(sici)1096-9896(199910)189:2<150::aid-path451>3.0.co;2-g

Cited by 58 publications

(21 citation statements)

References 47 publications

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“…This greatly hampers comparative gene expression studies and necessitates selective procurement of both normal and malignant ductal epithelial cells from within complex tissues. Laser capture microdissection (LCM) is a recently described method for isolating pure cell populations (Bonner et al, 1997;Sirivatanauksorn et al, 1999). We have used a Pixcell II system (Arcturus, Sunnyvale, USA) to obtain pure neoplastic and normal duct cells in order to investigate the molecular changes that accompany malignant transformation of this epithelial tumour type.…”

Section: Introductionmentioning

confidence: 99%

Expression profiling of microdissected pancreatic adenocarcinomas

et al. 2002

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Section: Introductionmentioning

confidence: 99%

Expression profiling of microdissected pancreatic adenocarcinomas

et al. 2002

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“…The other used a mixture of RNAs from chronic pancreatitis and health pancreas as the driver population in representational di erence analysis (Gress et al, 1997). Recently, the laser-capture microdissection technique for procuring individual components of complex tissues has been designed (Emmert-Buck et al, 1996;Sirivatanauksorn et al, 1999), but has proven to be labour-intensive and time-consuming. In addition, the amount of material generated by microdissection is limited and, without ampli®cation procedures, usually not su cient for gene expression studies.…”

Section: Introductionmentioning

confidence: 99%

Gene expression profiles of pancreatic cancer and stromal desmoplasia

Efthimiou²,

et al. 2001

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Gene expression studies were undertaken in normal pancreas and pancreatic adenocarcinomas to determine new candidate genes that can potentially be used as markers of the disease. The characteristic desmoplastic stromal reaction of pancreatic adenocarcinoma greatly hampers expression studies in this tumour type, and usually necessitates time-consuming tissue microdissection for enrichment of the tumour cell population. We show that ®ne needle aspiration of cancer provides a fast and e cient way of obtaining samples highly enriched in tumour cells with su cient yields of RNA. Using Atlas cancer cDNA arrays with 588 cancer-related genes, we describe gene expression pro®les of normal pancreas, bulk pancreatic tumour tissues and pancreatic tumour aspirates containing more than 95% tumour cells. Analysis of bulk tissue specimens revealed di erentially expressed genes belonging predominantly to the stromal component of the tumour. This contrasted with the results obtained from tumour-cell enriched samples. Several genes already described in pancreatic cancer (caspase 8, TIMP1, CD9, IL-13) were also di erentially expressed in our study. Furthermore, we found dysregulated expression of genes not previously associated with pancreatic adenocarcinoma, such as Rac 1, GLG1, NEDD5, RPL-13a, RPS9 and members of the Wnt5A gene family. In summary, we present a panel of genes newly identi®ed in the pathogenesis of pancreatic adenocarcinoma and demonstrate that ®ne needle aspirates of the tumour mass are a convenient source of material for gene expression studies in tumours accompanied by desmoplastic reactions. Oncogene (2001) 20, 7437 ± 7446.

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“…They have been shown to isolate precisely single cells or cell clusters under optical control for use in several downstream applications for DNA, RNA, and protein analysis. [5][6][7] In particular, the fragile mRNA can be obtained in a quality that is even suitable for construction of cDNA libraries or array hybridization.…”

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confidence: 99%

cDNA Array Hybridization after Laser-Assisted Microdissection from Nonneoplastic Tissue

Fink

Kohlhoff

Stein

et al. 2002

The American Journal of Pathology

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Differential gene expression can be investigated effectively by cDNA arrays. Because tissue homogenates result inevitably in an average expression of a bulk of different cells, we aimed to combine mRNA profiling with cell-type-specific microdissection. Using a polymerase chain reaction (PCR)-based preamplification technique, the expression profile was shown to be preserved. We modified the existing protocol enabling to apply the total amount of extracted RNA from microdissected cells. A mean amplification factor of nearly 1000 allowed to reduce the demand of initial RNA to ϳ10 ng. This technique was used to investigate intrapulmonary arteries from mouse lungs (ϳ500 cell equivalents). Using filters with 1176 spots, three independent experiments showed a high consistency of expression for the preamplified cDNAs. These profiles differed primarily from those of total lung homogenates. Additionally, in experimental hypoxia-induced pulmonary hypertension, amplified cDNA from intrapulmonary vessels of these lungs was compared to cDNA from vessels dissected from

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Laser-assisted microdissection: applications in molecular pathology

Cited by 58 publications

References 47 publications

Expression profiling of microdissected pancreatic adenocarcinomas

Expression profiling of microdissected pancreatic adenocarcinomas

Gene expression profiles of pancreatic cancer and stromal desmoplasia

cDNA Array Hybridization after Laser-Assisted Microdissection from Nonneoplastic Tissue

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