Flow cytometry-based systems and methods for detecting microbes

Abstract

In various embodiments, the present disclosure describes methods and systems for detecting microbes in a sample. The methods are generally applicable to quantifying the number of target bacteria in a sample counted from a detection region of a flow cytometer histogram. The detection methods can be employed in the presence of other microorganisms and other non-target microbe components to selectively quantify the amount of a target microbe. The methods are advantageous over those presently existing for testing of foodstuffs and diagnostic evaluation in their speed, accuracy and ease of use. Various swab collection devices and kits useful for practicing the present disclosure are also described herein.

What is claimed is: 
     
         1 . A method for optimizing the performance of a flow cytometer, said method comprising:
 a) increasing a sensitivity of at least one detection channel on the flow cytometer by increasing a gain on the at least detection one channel;   b) assigning a signal threshold values for each at least one detection channels;   c) subjecting particles to flow cytometer analysis;   d) collecting raw data from the flow cytometer for a time range; wherein the time range comprises a plurality of intervals; and wherein the raw data comprises signals and non-signals for each at least one detection channel; and   e) analyzing the raw data from each of the plurality of intervals to provide processed data; wherein analyzing comprises: eliminating raw data from each of the plurality of intervals in which the signals do not exceed the assigned signal threshold for each at least one detection channel; and selecting raw data from each of the plurality of intervals in which the signals do exceed the assigned signal threshold for each at least one detection channel.   
     
     
         2 . The method of  claim 1 , wherein increasing the sensitivity of at least one detection channel further comprises increasing a photomultiplier tube voltage. 
     
     
         3 . The method of  claim 1 , further comprising increasing the gain of the at least one detection channel. 
     
     
         4 . The method of  claim 1 , further comprising increasing both the gain of the at least one detection channel and increasing a photomultiplier tube voltage. 
     
     
         5 . The method of  claim 4 , further comprising subjecting the sample to serial gating. 
     
     
         6 . The method of  claim 1 , wherein the detection channel is FSC 
     
     
         7 . The method of  claim 1 , wherein the detection channel is SSC 
     
     
         8 . The method of  claim 1 , wherein the detection channel is FL-1. 
     
     
         9 . The method of  claim 1 , wherein the detection channel is FL-2. 
     
     
         10 . The method of  claim 1 , wherein the detection channel is FL-3. 
     
     
         11 . The method of  claim 1 , wherein the detection channel is FL-4. 
     
     
         12 . The method of  claim 1 , wherein the method for optimizing the performance of the flow cytometer further comprises panel equivalent target identification. 
     
     
         13 . The method of  claim 1 , wherein the method for optimizing the performance of the flow cytometer further comprises off target triangulation. 
     
     
         14 . The method of  claim 1 , wherein the at least one detection channel is a plurality of detection channels selected from the group of FSC, SSC, FL-1, FL-2, FL-3 and FL-4. 
     
     
         15 . The method of  claim 14 , wherein analyzing includes eliminating raw data in which the signals do not exceed the assigned signal threshold for any of the plurality of detection channels. 
     
     
         16 . The method of  claim 14 , wherein analyzing includes examining refractive properties of the particles. 
     
     
         17 . The method of  claim 1 , wherein the particles are individual cells, microbes, beads, quantum dots, gold particles quantum spheres or a combination thereof.