Rapid antibiotic susceptibility testing by tracking sub-micron scale motion of single bacterial cells

A method for rapid antibiotic susceptibility testing by tracking sub-micron scale motion of single bacterial cells comprising:

• adding different doses of at least one antibiotic to different ones of a plurality of wells in a multi-well glass slide, each well having a surface;
• adding portions of a biological sample with live bacteria to the plurality of wells, incubating bacterial cells from the live bacteria to tether a plurality of bacterial cells onto one of the surfaces to create tethered bacterial cells;
• operating a light microscopy apparatus to image each of the plurality of wells including imaging the tethered bacterial cells;
• operating a camera to obtain images of the tethered bacterial cells from the light microscopy apparatus;
• tracking the sub-micron motion of images of the tethered bacterial cells;
• analyzing bacterial motion of tethered cells in different wells at the different doses;
• operating a processor to apply an image analysis process to the images to quantitate the motions of the tethered bacterial cells to detect changes in the motions of the tethered cells;
• storing the motion changes in memory;
• and
• operating a processor to perform statistical analysis on a population of cells for each antibiotic dose to generate an antibiotic dose curve proportional to the motion changes, wherein the antibiotic dose curve plots data including a decrease in movement over time indicating a proportional effectiveness of an antibiotic applied to a well.

A method for rapid antibiotic susceptibility testing by tracking sub-micron scale motion of single bacterial cells comprising:

• adding different doses of at least one antibiotic to different ones of a plurality of wells in a multi-well glass slide, each well having a surface;
• adding portions of a biological sample with live bacteria to the plurality of wells, incubating bacterial cells from the live bacteria to tether a plurality of bacterial cells onto one of the surfaces as tethered bacterial cells, where the tethered bacterial cells are incubated with APTES in Ethanol on the surface for up to 15 seconds;
• operating a light microscopy apparatus to image each of the plurality of wells including imaging the tethered bacterial cells;
• operating a camera to obtain images of the tethered bacterial cells from the light microscopy apparatus;
• tracking the sub-micron motion of images of the tethered bacterial cells;
• analyzing bacterial motion of tethered cells in different wells at the different doses;
• operating a processor to apply an image analysis process to the images to quantitate the motions of the tethered bacterial cells to detect changes in the motions of the tethered cells;
• storing the motion changes in memory;
• and
• operating a processor to perform statistical analysis on a population of cells for each antibiotic dose to generate an antibiotic dose curve proportional to the motion changes, wherein the antibiotic dose curve plots data including a decrease in movement over time indicating a proportional effectiveness of an antibiotic applied to a well.