Instrument and system for rapid microorganism identification and antimicrobial agent susceptibility testing
A system for automated microorganism identification and antibiotic susceptibility testing comprising a reagent cartridge, a reagent stage, a cassette, a cassette, stage, a pipettor assembly, an optical detection system, and a controller is disclosed. The system is designed to dynamically adjust motor idle torque to control heat load and employs a fast focus process for determining the true focus position of an individual microorganism. The system also may quantify the relative abundance of viable microorganisms in a sample using dynamic dilution, and facilitate growth of microorganisms in customized media for rapid, accurate antimicrobial susceptibility testing.
1. A method for focusing on a microorganism to be imaged, comprising:
providing prescan information wherein intensity weighting versus focus position is obtained for a plurality of focus positions of an objective;
projecting light from a light source through the objective at an image plane;
capturing a first image of reflected light with the objective at a first position;
capturing a second image of reflected light with the objective at a second position, wherein the first position is an out-of-focus position a first fixed distance from a first side of the image plane and wherein the second position is an out-of-focus position a second fixed distance from a second side of the image plane, opposite the first side;
computing intensity weightings of the reflected light for the first and second images;
calculating a focus position using the computed intensity weightings in association with the prescan information by mapping the computed intensity weightings onto the prescan information, wherein the prescan information includes the intensity weighting versus focus position on both the first and second sides of the image plane;
moving the objective to the calculated focus position; and
capturing an image of a microorganism with the objective in the calculated focus position, which allows an identification of the microorganism.
2. The method of claim 1 , wherein the light source is a first light source used for generating a reflection off of a glass and liquid interface at the image plane and wherein the capturing of the image is with a second light source.
3. The method of claim 2 , wherein the first light source is a colored light source designed to generate a colored reflection off the glass and liquid interface and the second light source is a white light source designed to capture the image of the microorganism.
4. The method of claim 1 , further including computing a center of the reflected light in the first image and comparing the center to a known center to determine an offset distance from the known center and only using the first image if the offset distance is within a predetermined limit.
5. The method of claim 4 , further including repeatedly moving the objective and re-capturing the first image until the offset distance is within the predetermined limit.
6. The method of claim 1 , wherein the projecting of the light creates a spot of light on the image plane that is detectable through greyscale analysis.
7. The method of claim 1 , wherein the calculating of the focus position includes adding a known offset to an intensity weighting computation.
8. The method of claim 1 , wherein the computing of intensity weightings includes obtaining a greyscale value for each pixel in the first image and multiplying its greyscale value by its position relative to a center point divided by a full-image sum intensity, which is each pixel's intensity value added together.
9. The method of claim 1 , wherein the prescan information further includes spot offset versus focus position for the plurality of focus positions of the objective.
10. A method for focusing on a microorganism to be imaged, comprising:
capturing a first image of reflected light with an objective at a first position;
capturing a second image of reflected light with the objective at a second position, wherein the first position is an out-of-focus position a first fixed distance from a first side of an image plane and wherein the second position is an out-of-focus position a second fixed distance from a second side of the image plane, opposite the first side;
computing intensity weightings of the reflected light for the first and second images;
calculating a focus position using the computed intensity weightings through mapping of the computed intensity weightings onto prescan information, wherein the prescan information includes a plurality of intensity weightings versus focus positions on both the first side and the second side of the image plane; and
moving the objective to the calculated focus position for capturing an image which allows for an identification of the microorganism.
11. The method of claim 10 , wherein the focus position is determined based on data associated with the first and second images with respect to the prescan information and the method further includes capturing an image of a microorganism with the objective in the calculated focus position.