RNA sampling from tissue sections using infrared laser ablation

被引：7

作者：

Wang, Kelin ^{[1
]}

Donnarumma, Fabrizio ^{[1
]}

Herke, Scott W. ^{[2
]}

Dong, Chao ^{[1
]}

Herke, Patrick F. ^{[2
]}

Murray, Kermit K. ^{[1
]}

机构：

[1] Louisiana State Univ, Dept Chem, 331 Choppin Hall, Baton Rouge, LA 70803 USA

[2] Louisiana State Univ, Coll Sci, Genom Facil, Baton Rouge, LA 70803 USA

来源：

ANALYTICA CHIMICA ACTA | 2019年 / 1063卷

基金：

美国国家科学基金会;

关键词：

Laser ablation; RNA; Extraction; Rat brain tissue; Bioanalyzer; qPCR; HIGH-QUALITY RNA; SINGLE-STEP METHOD; GENE-EXPRESSION; CAPTURE MICRODISSECTION; MASS-SPECTROMETRY; MESSENGER-RNA; MICROARRAY; EXTRACTION; ACID; DNA;

D O I：

10.1016/j.aca.2019.02.054

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

RNA was obtained from discrete locations of frozen rat brain tissue sections through infrared (IR) laser ablation using a 3-mu m wavelength in transmission geometry. The ablated plume was captured in a microcentrifuge tube containing RNAse-free buffer and processed using a commercial RNA purification kit. RNA transfer efficiency and integrity were evaluated based on automated electrophoresis in microfluidic chips. Reproducible IR-laser ablation of intact RNA was demonstrated with purified RNA at laser fluences of 3-5 kJ/m(2) (72 +/- 12% transfer efficiency) and with tissue sections at a laser fluence of 13 kJ/m(2) (79 +/- 14% transfer efficiency); laser energies were attenuated similar to 20% by the soda-lime glass slides used to support the samples. RNA integrity from tissue ablation was >90% of its original RIN value (similar to 7) and the purified RNA was sufficiently intact for conversion to cDNA and subsequent qPCR assay. (C) 2019 Elsevier B.V. All rights reserved.

引用

页码：91 / 98

页数：8

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