Wright-Giemsa staining

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Method synopsis

Blood slide preparation and microscopic examination for haemosporidian parasite identification using Wright-Giemsa staining and compound microscopy with voucher image verification.

Synthesized from method descriptions across 5 papers using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Assessing the sensitivity of foraging and vigilance to internal state and environmental variables in yellow-bellied marmots (<i>Marmota flaviventris</i>). (2016), Behavioral Ecology and Sociobiology. Implementations in other papers (listed below) may differ.

1. Collect blood and fecal samples
   Blood samples of 0.5-2.5ml were collected from the femoral vein and immediately placed into heparinized tubes on ice for transport to lab where blood smears were made. Fresh fecal samples were collected in self-sealing plastic bag on ice for fecal glucocorticoid metabolite and intestinal parasite assays. Samples were collected within 2 weeks of behavioral observation date.
   Quantities: 0.5-2.5ml blood per individual, fecal samples from each captured individualDuration: During processing at captureConditions: Samples immediately placed on ice for transport
   Equipment: heparinized tubes, self-sealing plastic bags, ice
2. Process blood samples for parasite screening
   One drop of fresh blood was used to make thin film smears. Slides were stained and fixed using a Fisher Scientific Hema 3 stain kit and scored for presence or absence of Trypanosoma lewisi under microscope at 40x magnification. If no Trypanosoma lewisi were observed in 30 min, sample was scored as parasite free.
   Quantities: One drop blood per thin film smear, 30 min maximum screening time per sampleDuration: 30 minutes maximum per sample screeningConditions: 40x magnification microscope examination
   Equipment: Fisher Scientific Hema 3 stain kit, microscope

Typical Equipment

• sterile 30 gauge lancets
• micro-capillary tubes
• microscope slides
• alcohol fixative
• Wright-Giemsa stain
• compound microscope
• oil-immersion 100x lens
• digital imaging system

Output Measurements

• parasite presence/absence
• parasite species identification
• infection status

Papers Using This Protocol (5)

Species Studied With This Protocol

Places Where Used