The cumbersome restrictions of the suit, helmet, and gloves make many normal tasks and motions difficult or impossible. We will try various tools and methods to accomplish a set of both generalized and experiment-specific tasks that are routinely used in field geomicrobiology, mineralogy, and ecological studies.

Activities include the following, going from lesser to greater fine motor skills:

1. Simple hand samples collected in sterile Whirlpaks
   
   
2. Comparison of different sizes and types of forceps for manipulating small samples
   
   
3. Flame-sterilizing small tools with ethanol and lighter while not setting gloves on fire.
   
   
4. Gluing tiny samples onto SEM stubs
   
   
5. Screwing and unscrewing small culture tubes and inoculating media with sample.
   
   
6. Exoenzyme procedure: i.e. placing damp(fragile), sterile impregnated filter papers on surfaces, positioning sterile foil packets over them, putting in alkaline reaction stop solution, and retrieving the filters untorn.

Kelly Snook and Frank Schubert will test the first procedure during a full sim EVA to a desert varnish site previously scouted by Frank. Ephie Morphew and Sam Burbank will test all of the remaining procedures during a full sim EVA at another site.

Details of Procedures:

1. Sterile Whirlpaks will be opened by pulling on the two yellow tabs on the bag. Holding the outside of the bag, place the inside of the bag over specimen rock or chunk of material and scoop into the bag by squishing the bag around the object, turning bag over to upright position and letting material drop in. Twirl bag shut and bend in ends.
   
   
2. Take a selection of different forceps instruments and test the ability to pick up rocks of various sizes and shapes, small pieces of loosely consolidated materials, lichens, surface scrapings and mud patterns. Note the difficulty of wielding various tools and other drawbacks or advantages with the different sizes and styles. Will forceps in both hands help in manipulation? Do two people function better in manipulation than one?
   
   
3. Flame sterilizing is easier done with two people in almost all circumstances. Person 1: Dip business end of tool to be sterilized into small bottle of ethyl alcohol (ethanol) that you are holding. Person 2: Apply lighter, torch or other ignition device produced flame to the tool to light the alcohol. When using the tool first, repeat several times. Repeat between procedures that either touch different samples or are placed into different media or solutions, or anytime that a tool is accidentally brushed or touched to a non-sterile surface.
   
   
4. SEM stubs are small aluminum discs with a stem onto which samples are affixed for later coating with metal and analysis in scanning electron micrography. Usually needlepoint forceps are used to delicately manipulate the sample onto the disc. Conventional model glue (e.g. Duco Cement) is applied in a small drop to the disc and the sample is placed on it. Care must be taken to let the glue dry prior to packing or shaking the sample vials.
   
   
5. A variety of screwcap culture tubes are used to collect living organism samples in attempts to enable them to grow in captivity for further study. There are many kinds of media (bug food) that we use for various purposes. First, just to see if we can get anything to grow we use different kinds of media in hopes that something we offer will suit someone that we have captured. In other cases, we start by knowing that we want organisms that can do a particular type of metabolism. We make media that can indicate the presence of a byproduct of the metabolism that is of interest, e.g. by changing color, liquefying, or producing gas. Lastly, enrichment cultures are those that attempt to capture organisms that live by using a particular substance. We make bug food that contains only that substance. Whomever grows on it is suspected of being able to use that substance upon future verification experiments. Screwcap tubes are small and it is critical to keep the lip of the tube untouched and sterile. Samples are either plucked with sterile forceps, scraped with a sterile knife blade, collected with a sterile inoculating loop, or scooped with a small sterile metal spatula. The material is then placed in the tube and the tube is immediately capped. This procedure will be challenging with gloves on.
   
   
6. I have developed imprint exoenzyme assays to detect the presence and rough quantification of surface activities of organisms on rocky surfaces. All organisms exude substances into their environments. Frequently, these are enzymes that are characteristic of a particular metabolic process or other chemical transformation carried out by the organisms. We use a variety of substrates that various bug enzymes can act on. These are tagged with a chemical functional group that is not fluorescent when attached to the main substrate molecule, but becomes fluorescent when cleaved from that substrate by the appropriate enzyme if it encounters it on the sample surface. The way we usually do this is to have whirlpak bags with filter paper circles impregnated with the tagged substrates. Flame sterilized forceps are used to pull these out and place them on the surfaces. Pre-sterilized aluminum foil sheets are pressed over the filter to make it conform to the rock surface. These are left in place from 10 minutes to many hours depending upon the environmental conditions at a given site. Then the foil is lifted off, the filter paper is manipulated with forceps into an alkaline solution that stops the reaction. The filter is put on the foil, wrapped for protection and then returned to the lab for further processing and inspection with an epifluorescent microscope.