Quantum Microbiology™

Summary Points

• Quantum Microbiology™ (QM) is Accelr8's proprietary process for implementing single-cell microbiology methods.
• This page shows a demonstration of QM principles using images from a BACcelr8r™ research prototype.
• Sequence of images shows how images of immobilized cells, combined with appropriate assays, provide cellular-level data for rapid pathogen identification and antibiotic resistance analysis.
• The image sequence begins after sample preparation and introduction into a BACcelr8r cassette.
• This demonstration shows how QM methods will ultimately enable the BACcelr8r to deliver pathogen identification in 2 hours or less, rough antibitype identification in 4 hours or less, and complete antibiotic susceptibility testing in 8 hours or less.
• Complete analysis times are not fixed, but may be shorter when organisms prove susceptible to one or more fast-acting drugs. The system will deliver reports at the time data become statistically significant.

Analytical Image Sequence

The following sequence of image frames from the BACcelr8r research prototype's microscope shows the sequence of Quantum Microbiology™ (QM) assays. The sequence begins upon sample introduction into the cassette, after 30 minutes of specimen preparation (elapsed time is 30 minutes since starting sample prep). The right-hand column lists the time required for the process step and the cumulative time elapsed (parentheses) since starting sample prep. Times are examples from a specific run, and vary with different samples and processes.

Numbers in the lower-right of the single-clone images are the unique identification number of the clone. Each cassette has its own unique serial number, and each clone number encodes its flowcell identity. A database record tracks its spatial location within a field of view, indexed to an optical fiducial mark (not visible in the magnified images shown).

CAPTURE: Electrical field concentrates bacteria at the flowcell surface where they bind to a capture coating. The digital microscope focuses on the surface plane. Bacteria scatter the source light and appear as white specks.
(Partial field of view) Sample suspension enters the flowcell region. Few bacteria (bright specks) are near the surface		0 min (30 min)
End of capture period. All bacteria concentrated and captured on the surface. Electrical field stops.		+5 min (35 min)
SPECIES ID: Expose bacteria to various stains and other agents that indicate each organism's species. Image analyzer reads responses and color-codes each bacterial cell by type of response.
(Zoom in) Single progenitor cell on the surface, follow through the remaining steps. Add first ID reagent mix into the flowcell. Begin tracking growth.		+0 min (35 min)
Negative property (no stain) on Gram channel. Growing cell shows rod bacillus morphology.		+10 min (45 min)
Negative ID with first ID wave. Growth continues. Start second ID wave (of 2).		+0 min (45 min)
Positive ID on Klebsiella pneumoniae channel (green). Growth continues.		+10 min (55 min)
If necessary, allow additional growth for entire sample to meet criteria. Measure growth rate for each cell.		+35 min (90 min)
Report counts for each species (species response, Gram, morphotype) and growth rates.	REPORT Counts per species, morphology, Gram	+0 min (90 min)
ROUGH ANTIBIOTYPE ID: Identify each clone's major strain category, assigned by responses to key antimicrobial indicator agents.
Acquire images during exposure. Find each cell's growth or death in each interval. This clone not dead yet but growth stopped.		+10 min (100 min)
Later interval, count one cell killed in this clone (yellow or orange mortal stain channel positive, one cell), this interval, this agent.		+20 min (120 min)
Later interval, count second cell killed in this clone (mortal stain channel positive, one new cell), this interval, this agent.		+20 min (140 min)
Further images of this clone add no new information. Continue until report criteria met for all clones, then deliver report.		+70 min (210 min)
Report counts of major antibiotypes per species, count exceptions within species, for all clones and agents.	REPORT Counts per species, rough antibiotype ID	+0 min (210 min)
ANTIBIOTIC RESISTANCE ANALYSIS: Expose organisms to library of antibiotic drugs (one per flowcell). Measure the responses of each clone using a mortal stain (positive when cell dies). Continue to measure growth of unstained clones.
(Zoom out) Continue to measure growth and kill in all clones across all flowcells for all antimicrobials. View: back to the start of antibiotic delivery.		(replay)
Later image. Green cells have not yet died. Yellow and orange cells have died.		+1-5 hrs (~4-8 hrs)
Report time-kill statistics for each adequate agent. Call susceptibilities per species/drug. Count exceptions within species.	REPORT Time-kill, adequate agents. Susceptibility calls. Minority resistance counts.	+0 min (~4-8 hrs)