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Fluorescent Photo-conversion: A second chance to label unique cells.

ABSTRACT: Not all cells behave uniformly after treatment in tissue engineering studies. In fact, some treated cells display no signs of treatment or show unique characteristics not consistent with other treated cells. What if the "unique" cells could be isolated from a treated population, and further studied? Photo-convertible reporter proteins, such as Dendra2, allow for the ability to selectively identify unique cells with a secondary label within a primary labeled treated population. In the current study, select cells were identified and labeled through photo-conversion of Dendra2-transfected human Wharton's Jelly cells (hWJCs) for the first time. Robust photo-conversion of green-to-red fluorescence was achieved consistently in arbitrarily selected cells, allowing for precise cell identification of select hWJCs. The current study demonstrates a method that offers investigators the opportunity to selectively label and identify unique cells within a treated population for further study or isolation from the treatment population. Photo-convertible reporter proteins, such as Dendra2, offer the ability over non-photo-convertible reporter proteins, such as green fluorescent protein, to analyze unique individual cells within a treated population, which allows investigators to gain more meaningful information on how a treatment affects all cells within a target population.

SUBMITTER: Mellott AJ

PROVIDER: S-EPMC4406481 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

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Fluorescent Photo-conversion: A second chance to label unique cells.

Mellott Adam J AJ Shinogle Heather E HE Moore David S DS Detamore Michael S MS

Cellular and molecular bioengineering 20150301 1

Not all cells behave uniformly after treatment in tissue engineering studies. In fact, some treated cells display no signs of treatment or show unique characteristics not consistent with other treated cells. What if the "unique" cells could be isolated from a treated population, and further studied? Photo-convertible reporter proteins, such as <i>Dendra2</i>, allow for the ability to selectively identify unique cells with a secondary label within a primary labeled treated population. In the curr ...[more]

PMID: 25914756

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Project description:The main objective of this research was to determine the existence of relative age effect (RAE) in five European soccer leagues and their second-tier competitions. Even though RAE is a well-known phenomenon in professional sports environments it seems that the effect does not decline over the years. Moreover, additional information is required, especially when taking into account second-tier leagues. Birthdates from 1,332 first-tier domestic players from France, England, Spain, Germany and Italy and birthdates from 1,992 second-tier domestic players for the 2014/2015 season were taken for statistical analysis. In addition to standard statistical tests, the data were analyzed using econometric techniques for count data using Poisson and negative binomial regressions. The results obtained confirmed a biased distribution of birthdates in favor of players born earlier in the calendar year. For all of the five first-tier soccer leagues there was an unequal distribution of birthdates (France ?2 = 40.976, P<0.001; England ?2 = 21.892, P = 0.025; Spain ?2 = 24.690, P = 0.010; Germany ?2 = 22.889, P = 0.018; Italy ?2 = 28.583, P = 0.003). The results for second-tier leagues were similar (France ?2 = 46.741, P<0.001; England ?2 = 27.301, P = 0.004; Spain ?2 = 49.745, P<0.001; Germany ?2 = 30.633, P = 0.001; Italy ?2 = 36.973, P<0.001). Econometric techniques achieved similar results: estimated effect of month of birth, i.e., long-term RAE on players' representativeness, is negative (statistically significant at the 1% level). On average, one month closer to the end of the year reduces the logs of expected counts of players by 6.9%. Assuming this effect as linear, being born in the month immediately before the cut-off date (i.e., December/August), reduces the logs of expected counts of players by approximately 75.9%. Further, ID (index of discrimination, that is, the ratio between the expected counts of players born in the middle of the first and the twelfth month of the selection year) is 2.13 and 2.22 for the first- and second-tier, respectively. In other words, in the top five European first-tier and second-tier leagues, one should expect the number of players born in the first month of the calendar year to be twice the number of those born in the last month. The RAE in the second-tiers is the same as in the first-tiers, so it appears that there is no second chance for later born players. This reduces the chances to recover talented players discarded in youth simply because of lower maturity.

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Fluorescent Photo-conversion: A second chance to label unique cells.

Publications

Fluorescent Photo-conversion: A second chance to label unique cells.

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