Identification of expressed genes by laser-mediated manipulation of single cells

被引：342

作者：

Schütze, K ^{[1
]}

Lahr, G ^{[1
]}

机构：

[1] Acad Hosp Munchen Harlaching, Dept Med 1, Laser Lab & Mol Biol, Munich, Germany

来源：

NATURE BIOTECHNOLOGY | 1998年 / 16卷 / 08期

关键词：

gene analysis technology; in situ analysis; RT-PCR;

D O I：

10.1038/nbt0898-737

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We describe a rapid noncontact method for the capture of single cells or small tissue areas of any size or shape directly within the cap of a common microfuge tube. Prior to the laser-mediated transfer, the specimen is isolated by laser microbeam microdissection, forming a clear-cut gap around the selected area. Laser treatment does not impair subsequent RNA analysis. We have used this method to isolate a single cell from archival colon adenocarcinoma, and were able to detect point mutations within codon 12 of c-Ki-ras2 mRNA after nested RT-PCR analysis.

引用

页码：737 / 742

页数：6

共 37 条

[1]

Antinori S, 1996, HUM REPROD, V11, P2488

[2] RAS GENES [J].

BARBACID, M .

ANNUAL REVIEW OF BIOCHEMISTRY, 1987, 56 :779-827

[3]

BAUERLE D, 1996, LASER PROCESSING CHE, P420

[4]

Becker I, 1996, LAB INVEST, V75, P801

[5]

Bohm M, 1997, INT J ONCOL, V10, P131

[6]

Bohm M, 1997, AM J PATHOL, V151, P63

[7] Cell sampling - Laser capture microdissection: Molecular analysis of tissue [J].

Bonner, RF ;

EmmertBuck, M ;

Cole, K ;

Pohida, T ;

Chuaqui, R ;

Goldstein, S ;

Liotta, LA .

SCIENCE, 1997, 278 (5342) :1481-&

[8] 3 DIFFERENT MUTATIONS IN CODON-61 OF THE HUMAN N-RAS GENE DETECTED BY SYNTHETIC OLIGONUCLEOTIDE HYBRIDIZATION [J].

BOS, JL ;

VERLAANDEVRIES, M ;

JANSEN, AM ;

VEENEMAN, GH ;

VANBOOM, JH ;

VANDEREB, AJ .

NUCLEIC ACIDS RESEARCH, 1984, 12 (23) :9155-9163

[9] Laser beam microdissection in the diagnosis of cutaneous B-cell lymphoma [J].

Cerroni, L ;

Minkus, G ;

Putz, B ;

Hofler, H ;

Kerl, H .

BRITISH JOURNAL OF DERMATOLOGY, 1997, 136 (05) :743-746

[10] Fertilization of bovine oocytes induced solely with combined laser microbeam and optical tweezers [J].

ClementSengewald, A ;

Schutze, K ;

Ashkin, A ;

Palma, GA ;

Kerlen, G ;

Brem, G .

JOURNAL OF ASSISTED REPRODUCTION AND GENETICS, 1996, 13 (03) :259-265

← 1 2 3 4 →