High content screening technique

O Brien, P.J. (2008) Chapter 13 High content analysis of sublethal cytotoxicity in human HepG2 hepatocytes for assessing potential and mechanism for chemical and drug-induced human toxicity, in High Content Screening Science, Techniques and Applications (ed. S.A. Haney), John Wiley Sons, Hoboken, NJ,... 

Currently, two trends can be observed in the high throughput community miniaturization into the nanoliter dispensing regime and new high content screening (HCS) techniques. Small volume screening (either on 1536-well plates or the re-... 

The HPLC sugar assay was developed and validated as an efficient screening technique. As a result, some limitations may be encountered. Samples high in salt content may create difficulties, particularly interfering with glucose or sucrose. In some cases, some of the salt can be removed with an HPLC clean-up column or the use of a column different from the one specified (e.g., anion-exchange or calcium-loaded column). The preferred solution is to analyze such samples using GC instead of HPLC. 

Visual screening, facilitated by the color of the alkaloid, yielded highly productive cell lines of berberine (2/). The strong fluorescent properties of serpentine allowed the determination of its concentration in individual cells by flow cytometry, and subsequent sorting of the cells with high contents yielded a highly productive cell line (23). The same technique was used for berberine-containing cells (24). 

There is growing interest related to rapid screening and full characterization of the constituents of plants with medicinal properties. The high content in polyphenols accounts for in vitro and in vivo antioxidant activity of the extracts obtained from plants on the other hand, the high complexity of the samples extracted, depending on the method employed, may preclude complete resolution by conventional HPLC techniques. For this purpose, a comprehensive two-dimensional liquid chromatography (LCxLC) system, comprised of an RP-Amide first dimension and a partially porous octadecylsilica column in the second dimension, has been compared with a one-dimensional system (125). The chromatographic methods optimized in this research allowed the complete resolution and full characterization of polyphenols and xanthines in mate extracts. 

The requirement for drug screening is to have the ability to detect as wide a range of compounds as possible in as little sample (plasma/serum/whole blood, urine/vitreous humour, stomach contents/vomit or tissues) as possible at a high sensitivity but with no false positive(s). Ideally some sample should be left to permit confirmation of the results using another technique such as paper chromatography or TLC, and quantification of any poison(s) present to aid clinical interpretation of the results. 

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