Giemsa staining was used for the first two sporozoite challenge studies, and Field's stain in coplin jars for the later two studies. The thick film. Field Stain A and B, Preparation of the solution. Field stain consists of: Field stain A; Field stain B. Thin and thick blood films are made and fixed in methanol for. Rapid Field's Stain Method for Thin Blood Films. This is a modification of the original Field's stain to enable rapid staining of fixed thin films. It is a useful method for rapid presumptive species identification of malarial parasites.
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Do not heat fix. If patient specimen field stain blood, prepare routine thick and thin blood films. If patient specimen is solid such as corneal scrapings or a vaginal swabthe sample may be directly smeared field stain a clean glass slide.
Preparation of smears from Culture: Broths such as Modified Diamonds Medium, Cat. K02 or Acanthamoeba Broth, Cat. K should be mixed and an aliquot field stain be transferred to a sterile screw-capped tube or microcentrifuge tube.
Plates such as Non-Nutrient Agar, Cat. G should be scraped where feeding tracks are visible and directly smeared on a clean glass slide. Fix smears with 5 to10 drops of solution B, then field stain add equal drops of solution A. Add the stain powders Eosin and mix well.
Label the bottle Field stain B, field stain also write the date. Fill up two Coplin jars or wide-mouth bottles: Field Stain Field stain Blue stain.
Field Stain B Red stain.
Modified Field's Stain Kit
Make blood smear on a clean glass field stain and it is dried in the air. Fix in methanol for one minute or get Spray 'Easyfix'. Dry in the air. Neither managing clinicians nor field stain were aware of PCR data during the trial. Thick blood films Giemsa staining was used for the first two sporozoite challenge studies, and Field's stain in field stain jars for the later two studies.
Field stain - Wikipedia
The thick film was air dried in both methods. Field's field stain was applied by dipping the slide into Field's stain A for 3 seconds, then into tap water for 3 seconds with gentle agitationfield stain Field's stain B for a further 3 seconds and then washing gently in tap water to remove excess stain.
The slide was then air dried for at least 30 minutes. The lead microscopist examined slides produced by serial dilutions, blind to source. The increase in PCR product is quantitated by comparison field stain standard preparations of known parasite numbers. Results Sensitivity of blood films during sporozoite challenge The poisson distribution was used to calculate the likelihood of sampling a parasite within the blood volume examined in microscopy, at given parasite densities identified by PCR.
At low parasitaemias field stain was a discrepancy between the likelihood of diagnosis calculated by PCR readings, and the actual frequency of diagnosis at that density by thick film.
- Modified Field's Stain Kit
- Field stain Clinisciences
- Field Stain A and B, Preparation of the solution |
- Field stain
- Field Stain A and B, Preparation of the solution
This surprising finding suggested that a significant number of parasites were not visualized on a thick field stain despite being theoretically present in the original blood sample used to make the film.
Results did not vary according to staining protocol Giemsa or Field stain or by microscipist. A similar density threshold for reliable diagnosis of field stain by thick film examination is reported elsewhere [ 8 ].