Application and Progress of Raman Tweezers in Single Cells

College of Physics and Electronic Engineering, Guangxi Normal University, Guilin 541004, China
Chinese Journal of Analytical Chemistry (Impact Factor: 0.75). 05/2009; 37(5):758-763. DOI: 10.1016/S1872-2040(08)60105-X


Raman tweezers is a new optical technique that combines laser tweezers with Raman spectroscopy, which has the capability of studying single biological cells or organelles in nearly natured solution. Owing to its characteristics of noncontact, noninvasion, fast identification, and real-time detection, it has found application in improving the Signal Noise Ratio of Raman spectroscopy, optimizing the detection and sorting single cells or organelles, practicing the real-time detection of biochemistry dynamics process so as to make the activity and mechanisms of biomolecules understood deeply. The source, principle of Raman tweezers, and its optical application in biology are depicted and reviewed particularly in this article.

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    ABSTRACT: Laser tweezers Raman spectroscopy can help with observing and studying individual cells or organelles in a natural state for a relatively long period. In the present experiment, Laser tweezers Raman spectroscopy (LTRS) was used as a tool to report physiological metabolism such as cells growth and nucleic acid, proteins, lipid and glucose of a single active cold-adapted Aromatic hydrocarbons-degradating strains isolated from Antarctic Sea. After the Raman spectrum was collected and analyzed, the findings are as follows: Raman spectrum identified the components of a single cold-adapted Aromatic hydrocarbons-degradating strain and there were more proteins and carbohydrate produced during the Planococcus sp. NJ41 and Shewanella sp. NJ49 growth and degradation; but there was more lipid than the proteins produced during the Pseudoalteromonas sp. NJ289 growth and degradation; the amount of proteins produced by the strains corresponds with the production of degradation rate-limiting enzyme, and was also related to the capacity of low-temperature degradation of aromatic hydrocarbons.
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    ABSTRACT: Extensive research has been carried out in an effort to screen the oleaginous microorganisms. Here, Raman spectroscopy and laser tweezers were used to sort oleaginous yeast from mixed yeast cells. The preprocessing of subtracted background, 17 points S-G smoothing filter, polynomial fitting baseline correction and vector normalization were performed and the main features information of intracellular substances from the Raman spectroscopy of yeast cells was extracted by combining principal component analysis. Based on the distinguished composition of oleaginous yeast and non-oleaginous different yeast, a sorting model was established. The test yeast cell in optical trapping was distinguished real-time by the model referring to its Raman spectra. The cells distinguished as oleaginous yeast were collected by means of optical manipulation. The sorted oleaginous yeast cells were verified by microbial culture and Sudan black B test. The result illustrates that Raman spectroscopy combined with optical manipulation is an effective technique for sorting oleaginous yeast and other economic microorganisms.
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