ppat.1009262.g003.tif (333.89 kB)
Workflow from participant recruitment to image analysis.
figure
posted on 2021-02-01, 18:32 authored by Ryan Dinkele, Sophia Gessner, Andrea McKerry, Bryan Leonard, Ronnett Seldon, Anastasia S. Koch, Carl Morrow, Melitta Gqada, Mireille Kamariza, Carolyn R. Bertozzi, Brian Smith, Courtney McLoud, Andrew Kamholz, Wayne Bryden, Charles Call, Gilla Kaplan, Valerie Mizrahi, Robin Wood, Digby F. Warner(i) Recruitment of TB GeneXpert+ patients. (ii) One hour of bioaerosol production during tidal breathing and non-induced cough within the Respiratory Aerosol Sampling Chamber (RASC). (iii) Liquid capture of patient bioaerosol via Bertin Coriolis μ Biological Air Sampler. (iv) Bioaerosol concentration and staining with 100 μM DMN-trehalose during overnight (~16 hours) incubation at 37 oC. (v) Sample arraying within the nanowell device. (vi) Manual sample scanning and bacilli enumeration. (vii) Nanowell imaging (row 2 represents a zoomed in section from row 1). Columns represent 3 different patients. Bacilli not matching inclusion criteria are excluded from subsequent analysis. Scale bar, 5 μm.
