Abstract
Scientists have expressed continued interest in the development of microsensor technology that can indicate toxicity that occurs within a cell after a chemical challenge. One of the more useful approaches of microsensor technology is the application of fluorescent spectroscopy to indicate early-stage injury with optimal specificity and decreased background interference. If the toxicity is detected during the early onset period of injury, then the probability for therapeutic recovery is promising. There has been increasing interest in the use of infrared (IR) and near infrared (NIR) dyes as biological microsensors due to their fluorescent spectral characteristics. Three of the most essential characteristics are the ability to minimize background interference by extraneous biological matrices, to exhibit optimal molar absorptivity and quantum yields, to maintain chronic cellular homeostasis. Therefore, the present study was developed to determine if selected NIR dyes would distribute within isolated neutrophil populations without altering normal cellular homeostasis using fluorescent wavelength analysis. The results demonstrate that selected NIR dyes undergo internalization within target cell populations while maintaining cell viability and homeostasis. In addition, these dyes demonstrate changes in absorbance and fluorescence analysis after the immune cells were challenged with a stimulant. Moreover, critical cellular functions, such as degranulation and superoxide production were not compromised by the internalization of the NIR dyes. These data suggest that selected NIR dyes can undergo intracellular uptake within neutrophil cultures without altering the normal functional capabilities of the targeted cell population.
Highlights
The degree of environmental contamination has increased in recent years
The current study has demonstrated that the internalization of near infrared (NIR) micro-sensor dyes (NIR microsensor dyes 780 and 786) by isolated neutrophils did not result in a cellular lethality
Super Oxide Release (% Positive Control). These observations have demonstrated that the NIR micro-sensor dyes are not harmful once internalization within neutrophil populations has occurred
Summary
The degree of environmental contamination has increased in recent years Exposure to these contaminants could result in severe changes in normal cellular activity. Fluorescence is an energy phenomenon that involves the photoemission from excited molecules as their excited electrons return to their ground states [1, 2]. When the electron returns to the ground state, the absorbed energy is either emitted through a nonradiative process or a radiative one. Neutrophils were chosen as the in vitro cell model because they are an essential component of the body’s innate immune response They are a part of the first line of defense against antigenic invaders and they interact with many toxic substances or chemicals that may be distributed by the circulatory system to various tissues. If neutrophil activity is altered by any foreign invader, it would be beneficial to have the capability to monitor/indicate the development of cellular toxicity so that corrective therapy could be initiated
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