Borrelia Beacon 740, Fla and Control

Some fresh fingertip blood was stained with a molecular beacon for borrelia species.

Method: One drop of molecular beacon stain on centre of slide. Added small drop of fingertip blood and mixed. 50mm coverslip with grease (lithium high melting point wheel-bearing grease) on each corner placed and pushed down until whole area covered with sample. Stored in light-proof container with small vial of water to reduce dessication, long sides of coverslip sealed with adhesive at 6 to 12 hours. 48 hours after slide-prep heat cycle of 10 minutes @ 95C then slow cooling down to 50C which takes approximately 40 minutes.

Matched pairs of darkfield and fluorescence filtered images edited only for noise and size. One fairly bright autofluorescing subject was found on the control slide (shown below). The Fla beacon slide had a few more fluorescing subjects. The 740 beacon slide had many subjects mostly with weak to moderate fluorescence. These include small cysts of 1 - 3 microns though some larger subjects are seen. The latter raise the problem of autofluorescing small lymphocytes which with some methods/stains is a consideration. With the method used here this has not been a problem. The control slide had no autofluoresing small lymphocytes and the Fla slide (which can be treated as a second control) had a single potential candidate, making it highly probable that the 740 beacon has performed as expected by locking onto cells of borrelia spirochete morphological forms.

Molecular Beacon 740 Images

Molecular Beacon Fla images

Control Slide Images

Last update February 18th 2014

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