Research Techniques
The PDF files provided below give details on the techniques that have proven very reliable over many years in the field and laboratory. Also included are sources for supplies. Also shown below are some photographs to illustrate the techniques.
Making, staining, and storing blood smears.
See a movie of a smear being made
On the left below is shown three slides that have two smears made per slide. This is the method we use when traveling to field sites where large numbers of smears will be made. The slides shown in the picture on the left are fresh-made (not yet dipped into methanol). The top slide has one smear, the middle slide shows our practice when on field trips of putting two smears per slide, and the bottom slide is one typical for mammal blood with both a thick and thin smear.
The photo to the right shows some smears that have been stained with Giemsa. Three good slides are shown on the top. The ones staining pink are from gravid female lizards. Proteins used to produce egg yolk cycle through the blood and stain pink with Giemsa. The two lower slides show some poorly made smears. Note the “bubbles” on the left smear, second from the bottom. This is a result of grease on the slide.
Making and storing dried blood dots for genetic studies.
Storing dried blood is easy and should be done with every animal sampled. The left photograph shows a filter paper disk with several blood samples, the right photo shows the disk in a small plastic zip-lock bag with some silica gel to keep the disk dry. These disks can be kept at room temperature until returning to the lab, then kept in the freezer at -20 C.
Scanning slides, identifying and counting parasites.
How to catch lizards and take blood samples.
Catching and taking blood samples from lizards is easy, and described in the pdf file.
Noosing lizards can be very enjoyable, provided a properly made noose is used. We use tri-filament fishing line, but the pictures are shown here with heavy cord for illustration (such cord might be used to noose enormous Varanus lizards!). First, a knot is tied at the end of the line, then an overhand knot made. The long end of the string is pushed into the overhand knot, and then tightened. The picture on the lower left shows the finished noose. When a lizard is captured, it can be very quickly released by pulling on the “handle” of the noose (the knotted end).
Molecular methods: extraction DNA from dried blood dots, PCR, and sequencing
How to isolate microsatellite markers for malaria parasites