Laser Microdissection of Cells and Isolation of High-Quality RNA After

Higher plants are composed of different tissue and cell types. Distinct cells host different biochemical and physiological processes which is reflected in differences in gene expression profiles, protein and metabolite levels. When omics are to be carried out, the information provided by a specific cell type can be diluted and/or masked when using a mixture of distinct cells. Thus, studies performed at the cell- and tissue-type level are gaining increasing interest. Laser microdissection (LM) technology has been used to isolate specific tissue and cell types. However, this technology faces some challenges depending on the plant species and tissue type under analysis. Here, we show for the first time a LM protocol that proved to be efficient for harvesting specific tissue types (phloem, cortex and epidermis) from olive stem nodal segments and obtaining RNA of high quality. This is important for future transcriptomic studies to identify rooting-competent cells. Here, nodal segments were flash-frozen in liquid nitrogen-cooled isopentane and cryosectioned. Albeit the lack of any fixatives used to preserve samples’ anatomy, cryosectioned sections showed tissues with high morphological integrity which was comparable with that obtained with the paraffin-embedding method. Cells from the phloem, cortex and epidermis could be easily distinguished and efficiently harvested by LM. Total RNA isolated from these tissues exhibited high quality with RNA Quality Numbers (determined by a Fragment Analyzer System) ranging between 8.1 and 9.9. This work presents a simple, rapid and efficient LM procedure for harvesting specific tissue types of olive stems and obtaining high-quality RNA.

Download Full-text

Laser microdissection of conifer stem tissues: Isolation and analysis of high quality RNA, terpene synthase enzyme activity and terpenoid metabolites from resin ducts and cambial zone tissue of white spruce (Picea glauca)

BMC Plant Biology ◽

10.1186/1471-2229-10-106 ◽

2010 ◽

Vol 10 (1) ◽

pp. 106 ◽

Cited By ~ 58

Author(s):

Eric Abbott ◽

Dawn Hall ◽

Bjoern Hamberger ◽

Joerg Bohlmann

Keyword(s):

Enzyme Activity ◽

Picea Glauca ◽

Laser Microdissection ◽

White Spruce ◽

Terpene Synthase ◽

High Quality ◽

Resin Ducts ◽

Cambial Zone

Download Full-text

Some problems of objective-prism spectra classification

Symposium - International Astronomical Union ◽

10.1017/s0074180900053183 ◽

1966 ◽

Vol 24 ◽

pp. 51-52

Author(s):

E. K. Kharadze ◽

R. A. Bartaya

Keyword(s):

Short Wave ◽

High Quality ◽

Stellar Spectra ◽

Objective Prism

The unique 70-cm meniscus-type telescope of the Abastumani Astrophysical Observatory supplied with two objective prisms and the seeing conditions characteristic at Mount Kanobili (Abastumani) permit us to obtain stellar spectra of a high quality. No additional design to improve the “climate” immediately around the telescope itself is being applied. The dispersions and photographic magnitude limits are 160 and 660Å/mm, and 12–13, respectively. The short-wave end of spectra reaches 3500–3400Å.

Download Full-text

A non-cyanide method of electropolishing silver

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100107873 ◽

1978 ◽

Vol 36 (1) ◽

pp. 146-147

Author(s):

R. L. Lyles ◽

S. J. Rothman ◽

W. Jäger

Keyword(s):

Electron Microscopy ◽

Sulphuric Acid ◽

Methyl Alcohol ◽

Health Hazards ◽

Silver Alloy ◽

High Quality ◽

Silver Alloys ◽

Free Sample ◽

Specimen Quality ◽

Standard Techniques

Standard techniques of electropolishing silver and silver alloys for electron microscopy in most instances have relied on various CN recipes. These methods have been characteristically unsatisfactory due to difficulties in obtaining large electron transparent areas, reproducible results, adequate solution lifetimes, and contamination free sample surfaces. In addition, there are the inherent health hazards associated with the use of CN solutions. Various attempts to develop noncyanic methods of electropolishing specimens for electron microscopy have not been successful in that the specimen quality problems encountered with the CN solutions have also existed in the previously proposed non-cyanic methods.The technique we describe allows us to jet polish high quality silver and silver alloy microscope specimens with consistant reproducibility and without the use of CN salts.The solution is similar to that suggested by Myschoyaev et al. It consists, in order of mixing, 115ml glacial actic acid (CH3CO2H, specific wt 1.04 g/ml), 43ml sulphuric acid (H2SO4, specific wt. g/ml), 350 ml anhydrous methyl alcohol, and 77 g thiourea (NH2CSNH2).

Download Full-text