Core Outer Surface Biology Sampling:
The external surface of all received cores will be sampled and tested for the presence of adenosine triphospahte (ATP) (via bioluminescence) with the Lightning MVP instrument (manufactured by BioControl Systems Inc.) The Lightning MVP instrument uses a Luciferin-luciferase reagent to react with ATP to emit light. A cotton swab is swept across the surface of a core, then placed into a chamber where a reagent is added then the swab is exposed to a UV light. The reagent causes fluorescence in proportion to the amount of ATP present. The reading takes about 10s and it is measured in relative light units (RLU). The MVP instrument has a sensitivity of 15 Pico-grams of ATP. ATP is the energy mechanism for all living organisms so ATP luminometry is used as a screening instrument to determine the level of bacterial bioburden present on the cores. In MARTE it will provide a "quick look" assay for the presence of bacteria in cores.
Signs of Life Detection System (SOLID):
The instrument is a compact portable automated instrument that uses protein microarray technologies to detect microorganisms as well as their metabolic products. The aim is to detect any kind of biochemical compound (nucleic acids, proteins, polysaccharides, etc) using microarrays printed with antibodies or any other protein or molecule able to recognize and bind specifically to them. Molecular biology techniques allow fluorescent labeling of either the targets or the probes. A laser beam excites the sample and a CCD camera detects the bright spots. Core sections of scientific interest are sliced, powdered, and run through a series of molecular biology techniques using the SOLID instrument. SOLID performs an in-situ analysis of the core by using micro-arrays containing thousands of probes to detect organic compounds and whole cell characteristics from the powdered core samples, with a resolution better than PPB. Only 0.5 grams of powdered rock are required to conduct the procedure.
Biological Sample Extraction and Detection System (BSEDS) :
BSEDS uses Limulus Amebocyte Lysate (LAL) assay techniques to screen for biology and has as its main purpose to complement and make more robust, the existing biology detection system capability of the MARTE systems. The Limulus Amebocyte Lysate assay (LAL) is basically an aqueous enzyme extract from the blood cells of the horseshoe crab (Limulus Polyphemus) and is highly reactive to the bacterial endotoxins that are part of the lipopolysaccharide complex of gram negative bacteria and the beta glucan components of yeasts. When in the presence of the endotoxin, LAL initiates a cascade of enzyme activation steps that result in the proteolytic cleavage of an artificial peptide substrate. Upon cleavage this substrate releases p-nitroanaline, which is yellow in color and absorbs at 405 nm. The amount of endotoxin in the sample is quantified by the onset time of reaction and is measured using a laser diode at 405 nm coupled with a spectrophotometer. Hence shorter reaction times signify a higher amount of endotoxin in the sample and a standard endotoxin curve is used to help determine actual concentration of endotoxin in a given sample.
The system that will be used for the MARTE project is a modified version of commercially available handheld instrument called the Endosafe Portable Test System (PTS). The PTS will be coupled to an autonomous sample extraction system that will also provide the researcher with information about pH of the sample, specific ion concentration, and temperature. LAL uses a measure of endotoxin units per milliliter of sample (EU/ml). This measurement can be correlated back to the instrument calibration curve to obtain an approximate cell concentrating in the traditional measure of CFU/ml. The current sensitivity of the PTS system is 0.05 EU/ml which is equivalent to less than 1 CFU/ml. It is estimated that the PTS can detect as low at 1 cell per ml of sample.










