INTRODUCTION. Over the past 15 years since I joined support groups for patients with Lyme disease, an occasional and recurring topic of discussion has been breathing problems. These include a chronic cough and what some have described as having symptoms like persistent low-grade pneumonia. My recent microscopy study of sputum demonstrates the presence of significant numbers of spirochaetes showing varied morphology.
METHODS. Various methods to prepare slides were used. The simplest and a fairly effective method involves putting a small sample on a microscope slide with an added drop of DI water or saline. Wait a few seconds for the sample to absorb as much water as possible. This is carefully squashed with a 50mm glass coverslip until the sample spreads and becomes thin enough for high-power darkfield microscopy. Alternative 1. With the sample in a suitable sample collection tube with a cap, an equal volume of DI water or normal saline is added and the tube is shaken. This rapidly achieves full hydration of the sample which reduces its viscosity and improves handling. Once thinned, the sample can be centrifuged to eliminate bubbles. Alternative 2. Add to the sample an equal volume of: N-Acetyl L Cysteine 100mg/ml DI water or normal saline. Agitate vigorously and leave for a few minutes then centrifuge to eliminate bubbles. This is very effective for thinning the sample and is a first-stage in preparing a sputum sample for PCR.
RESULTS. Micrographs of the results can be seen here: https://www.flickr.com/photos/76898309@N08/albums/72157713091911953 The album of micrographs includes standard darkfield imaging of plain samples, giemsa stained and acridine orange stained samples. The latter includes matched pairs of filtered/unfiltered fields of view. The final set of compilation pictures demonstrate spirochaetes in varied forms, from cysts (curled into a ball inside a membrane) and partially open cysts, to fully formed spirochaetes.
For original unedited photos please contact me with the filename.
Peter Kemp. February 2020
The species of borrelia has yet to be determined as well as viability. Stay tuned.