US 5,675,517 A · Fluorescence spectral overlap compensation for high speed flow cytometry systems

Abstract

A fluorescence spectral overlap electronic compensation circuit for high speed flow cytometry systems is provided. In high-speed flow cytometry systems, such as systems with a pulse rise time up to 2 .mu.s, baseline restoration circuits may not adequately eliminate the DC offset of input signals, in which case, DC offset will result in errors after overlap compensation. In addition, analog spectral overlap compensation operations may result in signal distortions that are unacceptable when using a log amp whose output signal depends on the absolute value of its input signal. The disclosed fluorescence spectral overlap compensation circuit includes an adjustable DC offset compensation circuit to more accurately reduce the DC offset components of signals, and a half-wave rectifier for eliminating the signal distortions caused by spectral overlap compensation operations.

Claims21

Dependent claims

A preamplifier for use in a flow cytometry system, the preamplifier comprising:

a baseline restoration circuit coupled to receive a voltage-encoded signal, the baseline restoration circuit generating an estimated DC component responsive to the voltage-encoded signal;
and
an offset compensation circuit coupled to the baseline restoration circuit, the offset compensation circuit generating an offset signal indicating a voltage offset;
and
wherein the preamplifier is configured to attenuate the voltage-encoded signal based on the voltage offset and the estimated DC component to produce a DC compensated signal.

2 more elements

A data acquisition system comprising:

a detector disposed to detect particles having a first tag type, the detector generating an analog current which varies responsive to detecting particles having the first tag type;
a current-to-voltage converter coupled to the detector, the current-to-voltage converter receiving the analog current from said detector and generating the voltage-encoded signal responsive to the analog current;
a preamplifier comprising:
a baseline restoration circuit coupled to receive a voltage-encoded signal, the baseline restoration circuit generating an estimated DC component responsive to the voltage-encoded signal;
and
an offset compensation circuit coupled to the baseline restoration circuit, the offset compensation circuit generating an offset signal indicating a voltage offset;
and
wherein the preamplifier is configured to attenuate the voltage-encoded signal based on the voltage offset and the estimated DC component to produce a DC compensated signal;
an amplifier coupled to the preamplifier, the amplifier amplifying the DC compensated signal and generating an amplified signal;
an analog-to-digital converter coupled to the amplifier, the analog-to-digital converter generating digital data responsive to the amplified signal;
a computer coupled to the analog-to-digital converter, the computer categorizing the particles responsive to the digital data;
a second detector disposed to detect particles having a second tag type, the second detector generating a second analog current which varies responsive to detecting particles having the second tag type;
a reduced signal transmission circuit coupled to the second detector, the reduced signal transmission circuit receiving the second analog current and generating a reduced level signal, the reduced level signal being the second analog current reduced by a percentage;
an overlap compensation circuit coupled between the preamplifier and the amplifier, the overlap compensation circuit being coupled to the reduced signal transmission circuit, the overlap compensation circuit subtracting the reduced level signal from the DC compensated signal to produce an overlap compensated signal;
and
a half-wave rectifier coupled between the overlap compensation circuit and the amplifier, the half-wave rectifier receiving the overlap compensated signal and generating a rectified signal, the amplifier amplifying the rectified signal to produce the amplified signal.

13 more elements

A spectral overlap compensation system for use in a flow cytometry system, the spectral overlap compensation system comprising:

an overlap compensation circuit coupled to a first detector, said overlap compensation circuit receiving a first signal from said first detector, the overlap compensation circuit subtracting a second signal from the first signal to produce an overlap compensated signal, wherein the second signal is based on signals produced by one or more second detectors;
a half-wave rectifier coupled to the overlap compensation circuit, the half-wave rectifier receiving the overlap compensated signal and generating a rectified signal;
and
an amplifier coupled to the half-wave rectifier, the amplifier amplifying the rectified signal and generating an amplified signal.

1 more element

A data acquisition system for use in a flow cytometry system, comprising:

a first detector disposed to detect particles having a first tag type, the first detector generating a first analog current which varies responsive to detecting particles having the first tag type;
a first preamplifier coupled to the first detector, the first preamplifier receiving the first analog current from the first detector, the first preamplifier estimating a first estimated DC component of the first analog current based on the first analog current, the first preamplifier attenuating the first analog current based on a first voltage offset and the first estimated DC component to produce a first DC compensated signal;
a first overlap compensation circuit, the first overlap compensation circuit including
a first reduced signal transmission circuit coupled to the first preamplifier, the first reduced signal transmission circuit receiving the first DC compensated signal and generating a first reduced level signal, the first reduced level signal being the first DC compensated signal reduced by a first percentage, and
a first subtraction circuit coupled to the first preamplifier, the first subtraction circuit subtracting a second reduced level signal from the first DC compensated signal to produce a first overlap compensated signal;
a first half-wave rectifier coupled to the first overlap compensation circuit, the first half-wave rectifier receiving the first overlap compensated signal and generating a first rectified signal;
a first amplifier coupled to the first half-wave rectifier, the first amplifier amplifying the first rectified signal to generate a first amplified signal;
a first analog-to-digital converter coupled to the first amplifier, the first analog-to-digital converter generating digital data responsive to the first amplified signal;
a second detector disposed to detect particles having a second tag type, the second detector generating a second analog current which varies responsive to detecting particles having the second tag type;
a second preamplifier coupled to the second detector, the second preamplifier receiving the second analog current from the second detector, the second preamplifier estimating a second estimated DC component of the second analog current based on the second analog current, the second preamplifier Attenuating the second analog current based on a second voltage offset and the second estimated DC component to produce a second DC compensated signal;
a second overlap compensation circuit, the second overlap compensation circuit including
a second reduced signal transmission circuit coupled to the second preamplifier and to the first subtraction circuit, the second reduced signal transmission circuit receiving the second DC compensated signal and generating the second reduced level signal, the second reduced level signal being the second DC compensated signal reduced by a second percentage, and
a second subtraction circuit coupled to the second preamplifier and to the first reduced signal transmission circuit, the second subtraction circuit subtracting the first reduced level signal from the second DC compensated signal to produce a second overlap compensated signal;
a second half-wave rectifier coupled to the second overlap compensation circuit, the second half-wave rectifier receiving the second overlap compensated signal and generating a second rectified signal;
a second amplifier coupled to the second half-wave rectifier, the second amplifier amplifying the second rectified signal to generate a second amplified signal;
a second analog-to-digital converter coupled to the second amplifier, the second analog-to-digital converter generating digital data responsive to the second amplified signal;
a computer coupled to the first analog-to-digital converter and to the second analog-to-digital converter, the computer categorizing the particles responsive to the digital data generated by the first analog-to-digital converter and the second analog-to-digital converter.

14 more elements

Dependencies are inferred from the claim text.

Fluorescence spectral overlap compensation for high speed flow cytometry systems

Life of this patent

Fluorescence spectral overlap compensation for high speed flow cytometry systems

Life of this patent

Abstract

Claims21