procedure

FIELD COLLECTION

Locate a tree with approximately 0.1 square meters (1 square foot) of lichen coverage.
Using the thermometer, record the air temperature of the location under the shade of the tree (celsius).
Identify tree using a tree field guide or the Virginia Tech app.
Using the tablet and the air monitor, record the amount of particulate matter of the location (pm)
Take epicollect data of the air temperature, location, tree type, pictures of the tree, and pm of the area
Using a knife, collect a small (approximately 4x4 cm) piece of bark from the tree in a brown paper bag. Label the bag with tree type, location, date, initials, field number.
Using a scalpel, collect samples of lichen on the tree in a brown paper bag, approximately 1.5 grams total. Label the bag with tree type, location, date, initials, field number (all samples from the same tree have the same field number).
Record data on the data sheet.
Repeat steps 1-8 for each tree

pH (IN LAB)

Measure 5 mL of distilled water, use a pipette to transfer water and to measure 5 mL of water.
Transfer water to a 4 dram vial (21 mm x 70 mm diameter).
Using a knife cut the piece of bark into an approximately 2.5 gram sample (use scale to mass bark). Record the mass of the bark.
Place bark into the vial with the 5 mL of distilled water.
Label each vial appropriately (tree type, location, date, initials, field number).
Soak for 24 hours.
Use pH drops to test pH (5 drops for 5 mL of water).
Record pH of the sample.
Record data on the data sheet.
Repeat step 1-9 for each bark sample.

TARDIGRADES

From the collected lichen, measure 1.5 gram of lichen (use sterile container and a scale to measure 1.5 gram of lichen).
Measure 5 mL of distilled water, use a pipette to transfer water and to measure 5 mL of water.
Transfer the distilled water to a 4 dram vial (21 mm x 70 mm diameter).
Carefully transfer lichen to the vial, to ensure no lichen spills use a funnel.
Label each vial appropriately (tree type, location, date, initials, field number).
Soak for 24 hours.
Divide each sample into approximately 3 equal samples (0.5 grams of lichen) into 3 different petri dishes
Place petri dish under microscope at a magnification of 400x.
For 10 minutes, find as many tardigrades as possible by thoroughly inspecting 3 areas of the petri dish. Keep a tally mark of number tardigrades found during the 10 minute interval.
When the 10 minute interval is over, record the total number of tardigrades found for the sample on the data sheet.
Repeat step 1-8 for each lichen sample.