System for flexibly sorting particles

• US 5,550,058 A
• Filed: 12/15/1992
• Issued: 08/27/1996
• Est. Priority Date: 04/05/1991
• Status: Expired due to Fees

• Current Assignee: University of Rochester
• Original Assignee: University of Rochester
• Inventors: Leary, James F., Corio, Mark A.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Pyon, Harold Y.
• Application Number:
  US07/990,718
• Time in Patent Office: 1,351 Days
• Field of Search: 436/63, 436/800, 436/8, 209/906, 209/579, 209/3.1, 209/564, 422/82.01, 422/68.1
• US Class Current: 436/63
• CPC Class Codes: G01N 15/1427 with the synchronisation of components, a time gate for operation of components, or suppression of particle coincidences
• CPC Class Codes: G01N 15/1429 Signal processing
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: Y10S 209/906 Pneumatic or liquid stream feeding item

Abstract

This invention relates to a method and apparatus for flexibly controlling sorting decisions for a flow cytometer or similar instrument at a purity vs. yield ratio of sorted particles, particularly at high event rates. The system monitors distances between sequential objects to determine if they are in close enough proximity to be within a single resolution unit of the sorting device. The "friend" or "foe" status of neighboring events is considered by this system in dealing with detected "coincident" events. The system operates on information of the desirability to sort the object, i.e. droplet, based upon the properties of the individual object. A storage and retrieval method is provided for making the information available at the time a sort operation based on a sorting logic condition is performed. Accordingly, an easily reconfigurable mode of operation control is provided to allow customized sorting strategies, based on the needs of individual applications or experiments. This system may be included as an integral part of a flow cytometer/cell sorter or similar instrument and may also be provided as an outboard module to such systems.

Claims

1. A control system for sorting particles comprising:

a sensor for generating input signals for said particles;

timing circuitry receiving said input signals generated by said sensor and generating a trigger signal after a predetermined time interval for each of the input signals generated by said sensor;

inter-event counting circuitry for receiving the trigger signals generated by said timing circuitry and incrementing an inter-event counter in response to said input signals generated by said sensor received within said predetermined time interval;

inter-event storage circuitry for storing values of said inter-event counter;

sort decision storage circuitry for storing sort decision values for respective characteristics of said particles;

sort decision control circuitry receiving the trigger signal generated by said timing circuitry and thereby updating an availability level variable in response to the trigger signal generated by said timing circuitry;

said sort decision control circuitry generating a read storage signal for respectively reading sort decision values from said sort decision storage circuitry and values of said inter-event counter from said inter-event storage circuitry in response to the trigger signal generated by said timing circuitry;

comparing circuitry, coupled to said inter-event storage circuitry, for comparing said respective values of said inter-event counter with a predetermined value to thereby generate compared values when said respective values of said inter-event counter are less than said predetermined value; and

logic decision circuitry receiving said respective sort decision values from said sort decision circuitry, said availability level variable updated by said sort decision control circuitry, and said compared values generated by said comparing circuitry, and generating a sort pulse signal for said particles in response to the trigger signal generated by said timing circuitry whenever a selected yield/purity ratio which includes intermediate values between maximum yield and maximum purity occurs.

2. A system for sorting particles as in claim 1, which performs the steps of:

receiving detection signals from a sensor representing selected parameters of moving particles;

generating respective trigger signals and a respective individual sort decisions in response to the detection signals received from said sensor;

detecting an overlap condition of successive respective trigger signals in response to time separation between trigger signals of less than selected intervals and producing respective overlap detecting signals in response to the overlap condition;

determining a logic condition in response to a selected combination of the respective individual sort decisions and the overlap detecting signals; and

applying a sort pulse to the moving particles in response to the logic condition.

3. A system for sorting particles as in claim 2, wherein the value of the sort pulse applied to the moving particles is defined based on the logic condition.

4. A control system for sorting particles comprising:

a source of detection signals pertaining to selected parameters for respective particles generated in response to the respective particles passing a selected point;

a timing circuit which receives the detection signals and generates a respective trigger signal;

an overlap detection circuit, responsive to successive trigger signals generated by said timing circuit, to generate respective overlap signals in response to time separation of less than selected intervals;

a sort logic circuit, responsive to the trigger signals generated by said timing circuit, the detection signals and the overlap signals generated by said overlap detection circuit, to generate respective sort logic signals to be applied to the respective particles whenever a selected yield/purity ratio which includes intermediate values between maximum yield and maximum purity correspond to a selected logic condition.

5. A system for sorting particles as in claim 4, which performs the steps of:

receiving detection signals from a sensor representing selected parameters of moving particles;

generating respective trigger signals and a respective individual sort decisions in response to the detection signals received from said sensor;

detecting an overlap condition of successive respective trigger signals in response to time separation between trigger signals of less than selected intervals and producing respective overlap detecting signals in response to the overlap condition;

determining a logic condition in response to a selected combination of the respective individual sort decisions and the overlap detecting signals; and

applying a sort pulse to the moving particles in response to the logic condition.

6. A system for sorting particles as in claim 5, wherein the value of the sort pulse applied to the moving particles is defined based on the logic condition.

7. A system for sorting particles as in claim 4 in which the timing circuit further comprises an event timer circuit responsive to the trigger signal for measuring a time interval to when the sort logic signals generated by the sort logic circuit are applied to the respective particles and generating an event timing signal at the end of the time interval.

8. A system for sorting particles as in claim 7 in which the overlap detection circuit further comprises an overlap storage circuit including a plurality of latches for storing successive respective overlap signals and a coincidence detection circuit for comparing the successive overlap signals to the selected intervals of time separation.

9. A system for sorting particles as in claim 8 in which the sort logic circuit comprises:

a sort decision storage circuit, including a plurality of latches, for storing successive detection signals;

a sort value storage control circuit controlling the storing of the successive respective overlap signals in the overlap storage circuit and the successive detection signals in the sort decision storage circuit and generating an availability value in response thereto; and

a sort decision logic look-up table responsive to the availability value generated by the sort value storage control circuit, the successive respective detection signals and the successive respective overlap signals generated by the overlap detection circuit, to generate respective sort logic signals whenever a selected combination of the successive availability values generated by the sort value storage control circuit, successive respective detection signals and successive respective overlap signals generated by the overlap detection circuit occurs.

10. A system for sorting particles as in claim 9 further comprising a sort pulse definition circuit to generate the value of the respective sort logic signal to be applied to the particles.

11. A system for sorting particles as in claim 10 further comprising a sort pulse amplifier circuit to amplify the respective sort logic signals before applying the sort logic signals to the particles.