Borrelia burgdorferi + Borrelia garinii FITC

Some fresh fingertip blood was mixed with KPL's anti-borrelia spp. polyclonal FITC antibody (cat. no.02-97-91 Link to product page) AND Abcam's Anti-Borrelia burgdorferi garinii polyclonal FITC antibody (cat. no. ab20118 Link to product page).

Method: Approx 10 to 15uL of each fluorescent antibody stain (diluted 1:20 distilled water) on centre of slide. Added small drop of fingertip blood and mixed. 50mm coverslip with grease on each corner placed and pushed down until whole area covered with sample. At this point the sample is deep enough for continuous mixing of the sample with the stains whilst stored in a dark, humid box kept in a warm place. After 6 hours the grease holding-up the coverslip was pressed down firmly to produce a shallow sample and the long sides of the coverslip were sealed with adhesive. Then stored for 18 hours (24 hours total) before microscopy observations made over the following 24 hours.

Previous experiments with either of the stains had shown some interesting results with some agents clearly fluorescing far above background or autofluorescence levels. However, the staining used high concentrations to be effective suggesting that not all the flourescien conjugated antibodies were finding matched protiens in the sample. The results from the combined stains could be explained by agents in the sample binding different antibodies from both stains. If correct, this could indicate that the borrelia species in the sample is related to both Borrelia burgdorferi and Borrelia garinii but might not be an exact match for either. Alternative explanations could be that the borrelia morphology found in fresh or slightly aged (up to 24 hours) whole blood; or borrelia morphology found in persistent infection, only presents with certain protiens that some antibodies can attach to and these happen to have matched some present in the combined stains.

The Control Slide was searched for the most fluorescent subjects that could be found. Images at the bottom of the page show the generally modest to minimal autofluorescence of some lymphocytes and other agents. Exposure time/sensitivity for the Control Slide was 2 to 4 times the exposure levels required for the stained slides.

The images below show matched pairs of the same field of veiw of darkfield (the 'yellow' picture in each pair) and fluorescence filtered images (the 'green' picture in each pair) are unedited except for cropping and size reduction.

Control Slide:

 

Last update January 31st 2014

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