Evaluation circuit for an optoelectronic detector and method for recording fluorescence events

• US 9,377,405 B2
• Filed: 09/20/2012
• Issued: 06/28/2016
• Est. Priority Date: 09/30/2011
• Status: Active Grant

• Current Assignee: Carl Zeiss Microscopy GmbH (Carl-Zeiss-Stiftung)
• Original Assignee: Carl Zeiss Microscopy GmbH (Carl-Zeiss-Stiftung)
• Inventors: Mhler, Gunter, Schmidt, Dietmar, Holub, Oliver, Hauschild, Robert
• Primary Examiner(s): Porta, David
• Assistant Examiner(s): GUNBERG, EDWIN C
• Application Number:
  US14/348,261
• Publication Number:
  US 20140231675A1
• Time in Patent Office: 1,377 Days
• Field of Search: 250/459.1
• US Class Current: 1/1
• CPC Class Codes: G01J 1/44 Electric circuits for command of an exposure part G03B7/02
• CPC Class Codes: G01J 3/2889 Rapid scan spectrometers; Time resolved spectrometry
• CPC Class Codes: G01J 3/44 Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/0084 time-scale detection, e.g. strobed, ultra-fast, heterodyne detection

Abstract

An improved evaluation circuit which allows high sensitivity in an economical manner. For this purpose, a shift register having at least one data input, a clock input, a plurality of register stages and at least one data output is provided, wherein the output of the analog-to-digital converter is connected to the data input of the shift register. With a shift register, fluorescent light and scattered light can be temporally distinguished in an economical manner.

Claims

1. An evaluation circuit for evaluating the electric signals of an optoelectronic detector, comprising:

an analog-to-digital converter comprising;

an input configured to connect to an output of the detector;

an output configured to output a digital value;

a shift register comprising;

at least one data input;

a clock input;

a plurality of register stages; and

at least one data output;

wherein the output of the analog-to-digital converter is connected to the data input of the shift register and also to the clock input of the shift register, so that the contents of the shift register are shifted by one stage as a result of said digital value differing from zero.

2. The evaluation circuit according to claim 1, further comprising:

a delay circuit with variably adjustable time delay, the delay circuit comprising;

an input configured to receive a signal indicating a light pulse; and

an output configured to output the signal after a selected delay as a delayed signal;

wherein the output of the delay circuit is connected to the data input of the shift register such that, by outputting the delayed signal, a marking value is presented at the data input in addition to or instead of the digital value.

3. The evaluation circuit according to claim 2, further comprising:

a readout circuit comprising;

an input configured to receive a trigger signal;

at least one data input which is connected to the data output of the shift register; and

at least one sum output;

wherein the readout circuit is configured to read out the register stages when the trigger signal is present;

wherein the readout circuit is also configured to;

sum digital values stored before the marking value; and

/or

sum digital values stored after the marking value; and

wherein the readout circuit is further configured to output the summed values to the sum output.

4. The evaluation circuit according to claim 1, further comp rising:

at least one additional shift register; and

a switching circuit which, for a plurality of consecutive light pulses, is configured to alternately connect the shift registers to the analog-to-digital converter.

5. A microscope comprising:

a light source configured for pulsed operation;

at least one of;

a pulse generator configured to control the light source to emit light pulses; and

a pulse monitor configured to monitor the generated light pulses;

an optoelectronic detector; and

the evaluation circuit according to claim 1;

wherein the input of the analog-to-digital converter is connected to the output of the detector or is configured to be connected to the output of the detector by means of a switch.

6. The microscope according to claim 5;

wherein the evaluation circuit comprises;

a delay circuit with variably adjustable time delay, the delay circuit comprising;

an input configured to receive a signal indicating a light pulse; and

an output configured to output the signal after a presently selected delay as a delayed signal;

wherein the output of the delay circuit is connected to the data input of the shift register such that, by outputting the delayed signal, a marking value is presented at the data input in addition to or instead of the digital value; and

wherein the signal input of the delay circuit is connected to the pulse generator or to the pulse monitor.

7. The microscope according to claim 6;

wherein the delay circuit has a delay corresponding to a duration of the light pulses.

8. The microscope according to claim 6, further comprising:

operating means for setting the delay of the delay circuit.

9. The microscope according to claim 6;

wherein the evaluation circuit comprises;

a readout circuit comprising;

an input configured to receive a trigger signal;

at least one data input which is connected to the data output of the shift register; and

at least one sum output;

wherein the readout circuit is configured to read out the register stages when the trigger signal is present;

wherein the readout circuit is also configured to;

sum digital values stored before the marking value; and

/or

sum digital values stored after the marking value; and

wherein the readout circuit is further configured to output the summed values to the sum output;

wherein the evaluation circuit is connected via a data bus to a control unit configured to transmit detected fluorescence events; and

wherein the readout circuit is arranged in front of the data bus.

10. A microscope comprising:

a light source configured for pulsed operation;

the evaluation circuit according to claim 1;

at least one of;

a pulse generator configured to control the light source to emit light pulses; and

a pulse monitor configured to monitor the emitted light pulses;

an optoelectronic detector; and

a delay circuit with variably adjustable time delay, the delay circuit comprising;

an input configured to receive a signal indicating a light pulse; and

an output configured to output the signal after a presently selected delay as a delayed signal;

a deactivation circuit configured to occasionally, at least partially, deactivate the optoelectronic detector;

wherein the deactivation circuit is connected to the output of the delay circuit such that the optoelectronic detector is deactivated by outputting the delayed signal.

11. A method for recording fluorescence events by means of an optoelectronic detector, comprising:

radiating light pulses onto a sample by means of a light source;

capturing light from the sample by means of the optoelectronic detector;

converting the captured light into an electric signal and

feeding the electric signal to an analog-to-digital converter to generate a digital value;

outputting the digital value from the analog-to-digital converter; and

storing the outputted digital value in an input stage of a shift register;

wherein the method farther comprises at least one further step selected from the group consisting of;

summing digital values which are stored in the shift register before a marking value; and

summing digital values which are stored in the shift register after a marking value.

12. The method according to claim 11;

wherein contents of the shift register are shifted by one stage at the occurrence of a non-zero digital value.

13. The method according to claim 11;

wherein the contents of the shift register are shifted by one stage repeatedly after a predefined time, where the predefined time is provided by a clock generator having a predefined or predefinable clock frequency.