System and method for calculating a droplet delay time, and sorting device
The present application discloses a system and a method for calculating a droplet delay time in a liquid flow of a sorting device. The system includes: a first laser source configured to emit a first laser beam to a liquid flow of a sorting device, the first laser beam and the liquid flow intersecting at a first laser interrogation point; a second laser source configured to emit a second laser beam to the liquid flow, the second laser beam and the liquid flow intersecting at a second laser interrogation point; and a droplet delay time calculation unit configured to calculate, based on of the time when the particle in the liquid flow passes through the first and second laser interrogation point and the time when the particle passes through the second laser interrogation point, a first delay time, and calculate, a droplet delay time.
1 . A system for calculating a droplet delay time in a liquid flow of a sorting device, comprising:
a first laser source configured to emit a first laser beam to the liquid flow of the sorting device, the first laser beam and the liquid flow intersecting at a first laser interrogation point located in a nozzle system of the sorting device;
a second laser source configured to emit a second laser beam to the liquid flow, the second laser beam and the liquid flow intersecting at a second laser interrogation point located outside the nozzle system, wherein the second laser interrogation point is before a droplet separation point;
a first detector corresponding to the first laser source and configured to detect emission, in response to the first laser beam, of a particle in the liquid flow to determine the time when the particle passes through the first laser interrogation point;
a second detector corresponding to the second laser source and configured to detect emission, in response to the second laser beam, of the particle in the liquid flow to determine the time when the particle passes through the second laser interrogation point;
a processor programmed to calculate, on the basis of the time when the particle in the liquid flow passes through the first laser interrogation point and the time when the particle passes through the second laser interrogation point, a first delay time for the liquid flow flowing from the first laser interrogation point to the second laser interrogation point, and calculate, on the basis of the first delay time, a droplet delay time,
wherein the processor is further configured programmed to calculate a second delay time for the liquid flow flowing from the second laser interrogation point to the droplet separation point and calculate the sum of the first delay time and the second delay time as the droplet delay time; and
a camera device adapted to obtain an image of the liquid flow, the second laser interrogation point and the droplet separation point being within a pick-up range of the camera device, wherein processor is further programmed to calculate the second delay time by using a plurality of images of the liquid flow obtained by the camera device.
2 . The system according to claim 1 , wherein the processor is further programmed to calculate an average value of first measurement times of a plurality of particles as the first delay time,
wherein for each particle, the first measurement time refers to a difference between the time, determined on the basis of the detection of the first detector, when the particle passes through the first laser interrogation point and the time, determined on the basis of the detection of the second detector, when the particle passes through the second laser interrogation point.
3 . The system according to claim 1 , wherein the processor is further programmed to: for each of multiple types of particle sizes, calculate a first delay time for the particle size on the basis of first measurement times of a plurality of particles with the particle size, so that the first delay time is calculated to be different for different particle sizes,
wherein for each particle, the first measurement time refers to a difference between the time, determined on the basis of the detection of the first detector, when the particle passes through the first laser interrogation point and the time, determined on the basis of the detection of the second detector, when the particle passes through the second laser interrogation point.
4 . The system according to claim 3 , further comprising a particle size calculation unit configured to, for each particle, calculate the size of the particle on the basis of a pulse width of a signal obtained by the first detector detecting the emission, in response to the first laser beam, of the particle and a spot size of the first laser beam.
5 . The system according to claim 3 , wherein the processor is further programmed to create, on the basis of first delay times of the multiple types of particle sizes, a lookup table or an expression that characterizes a relationship between particle sizes and first delay times.
6 . The system according to claim 1 , further comprising a third laser source used for emitting a third laser beam to the liquid flow for fluorescence excitation; and
a light beam adjusting element configured to transmit the first laser beam and the third laser beam to the liquid flow along the same path,
wherein the first laser beam is also used for fluorescence excitation.
7 . The system according to claim 1 , wherein the sorting device is a flow cytometer.
8 . The system according to claim 7 , wherein the flow cytometer is a glass-pool-based flow cytometer;
the nozzle system comprises a glass pool and a nozzle; and
the first laser interrogation point is located in the glass pool.
9 . A sorting device, comprising the system according to claim 1 .
10 . A method for calculating a droplet delay time in a liquid flow of a sorting device, comprising:
respectively using a first laser source and a second laser source of the sorting device to emit a first laser beam and a second laser beam to a liquid flow of the sorting device;
calculating, on the basis of the time, determined on the basis of detection of a first detector of the sorting device, when a particle in the liquid flow passes through a first laser interrogation point located in a nozzle system of the sorting device and the time, determined on the basis of detection of a second detector of the sorting device, when the particle passes through a second laser interrogation point outside the nozzle system, a first delay time for the liquid flow flowing from the first laser interrogation point to the second laser interrogation point;
calculating a droplet delay time on the basis of the first delay time,
wherein the first laser beam and the liquid flow intersect at the first laser interrogation point;
wherein the second laser beam and the liquid flow intersect at the second laser interrogation point;
wherein the second laser interrogation point is before a droplet separation point;
wherein the first detector is configured to detect emission, in response to the first laser beam, of the particle in the liquid flow to determine the time when the particle pass through the first laser interrogation point; and
wherein the second detector is configured to detect emission, in response to the second laser beam, of the particle in the liquid flow to determine the time when the particle pass through the second laser interrogation point; and
calculating a second delay time for the liquid flow flowing from the second laser interrogation point to the droplet separation point, wherein the operation of calculating the droplet delay time comprises:
calculating the sum of the first delay time and the second delay time as the droplet delay time; and
calculating the second delay time by using a plurality of images of the liquid flow obtained by a camera device, wherein the second laser interrogation point and the droplet separation point are within a pick-up range of the camera device.
11 . The method according to claim 10 , wherein the operation of calculating the first delay time comprises: calculating an average value of first measurement times of a plurality of particles as the first delay time,
wherein for each particle, the first measurement time refers to a difference between the time, determined on the basis of the detection of the first detector, when the particle passes through the first laser interrogation point and the time, determined on the basis of the detection of the second detector, when the particle passes through the second laser interrogation point.
12 . The method according to claim 10 , wherein the operation of calculating the first delay time comprises: for each of multiple types of particle sizes, calculating a first delay time for the particle size on the basis of first measurement times of a plurality of particles with the particle size, so that the first delay time is calculated to be different for different particle sizes,
wherein for each particle, the first measurement time refers to a difference between the time, determined on the basis of the detection of the first detector, when the particle passes through the first laser interrogation point and the time, determined on the basis of the detection of the second detector, when the particle passes through the second laser interrogation point.
13 . The method according to claim 12 , further comprising: for each particle, calculating the size of the particle on the basis of a pulse width of a signal obtained by the first detector detecting the emission, in response to the first laser beam, of the particle and a spot size of the first laser beam.
14 . The method according to claim 12 , wherein the operation of calculating the first delay time further comprises: creating, on the basis of first delay times of the multiple types of particle sizes, a lookup table or an expression that characterizes a relationship between particle sizes and first delay times.
15 . The method according to claim 10 , further comprising: using a third laser source of the sorting device to emit a third laser beam to the liquid flow for fluorescence excitation, wherein the first laser beam is also used for fluorescence excitation, and wherein the first laser beam and the third laser beam are transmitted to the liquid flow along the same path by a light beam adjusting element of the sorting device.
16 . The method according to claim 10 , wherein the sorting device is a flow cytometer.
17 . The method according to claim 16 , wherein the flow cytometer is a glass-pool-based flow cytometer, the nozzle system comprises a glass pool and a nozzle; and the first laser interrogation point is located in the glass pool.