System for axial pattern analysis of multicellular organisms

A method for sorting multicellular organisms comprising the steps of:

• providing a population of test organisms, wherein each member of the population displays at least one spatially distinct, optically detectable, phenotypic characteristic;
• analyzing the arrangement of spatially distinct, optically detectable, phenotypic characteristics of each population member;
• and
• depositing members of the population based on the arrangement of spatially distinct, optically detectable, phenotypic characteristics.

An instrument for analyzing and selectively dispensing elongate multicellular organisms comprising:

• a source containing multicellular organisms in a fluid suspension;
• means for causing the fluid suspension to move in a direction of flow;
• means for aligning the elongate multicellular organisms relative to the direction of flow;
• a light source for producing an optical beam through which the elongate multicellular organisms pass after becoming aligned;
• a first optical detector for detecting light over a solid angle of at least 20 degrees and over a collection angle of approximately 0.0 to 6.0 degrees in the horizontal axis and approximately 17 degrees in the vertical axis for detecting passage of said organisms through said optical beam;
• and
• a fluid switch downstream of a point where said organisms pass through said optical beam, said switch responsive to the first optical detector to allow detected objects to pass to a sample container.

A method of selectively dispensing elongate multicellular organisms comprising the steps of:

• centering and orienting the sample objects in a flowing fluid stream;
• passing the fluid stream through a sensing zone;
• optically detecting the presence of a multicellular organism passing through the sensing zone by means of a light scatter sensor that has an acceptance angle of at least 20 degrees and over a collection angle of approximately 0.0 to 6.0 degrees in the horizontal axis and approximately 17 degrees in the vertical axis;
• creating a data representation of sequential optical characteristics of the multicellular organism comprising output signals from additional optical sensors;
• diverting at least some portion of the fluid stream with a switched fluid stream based on the data representation so as to collect ones of the multicellular organisms remaining in portions of the sample stream that were not diverted.

A process for analyzing elongate multicellular organisms by flow cytometry comprising the steps of:

• creating a population of test organisms wherein each member of the population displays a marker pattern, said marker pattern representing a plurality of spatially consistent first features spaced apart along a length of each organism and wherein each member of the population also displays at least one of a second feature modifiable or inducible when the population is subjected to a test treatment, each of said first and said second features being detectable through analysis with a flow cytometer;
• subjecting the population to a test treatment;
• analyzing members of the population with a flow cytometer equipped to process elongate multicellular organisms;
• detecting the marker pattern on the members analyzed;
• and
• using the detected marker pattern to determine status of the second feature on each of the members analyzed.

A process for preparing a model strain of elongate multicellular organisms intended for specialized flow cytometry analysis comprising the steps of:

• creating a marker strain of organisms wherein each member of the strain displays a marker pattern, said marker pattern representing a plurality of marker features spaced apart along a length of each organism and spatially consistent from member to member, said marker features being detectable through analysis with a flow cytometer;
• creating a test strain of organisms wherein each organism of the test strain displays at least one test feature modifiable or inducible when the test strain is subjected to a test treatment, said test features being detectable through analysis with a flow cytometer;
• and
• creating a model strain by combining the marker pattern from the marker strain with the test features from the test strain so that each organism of the model strain displays both the marker pattern and at least one test feature.