Luminescence measurement of biological samples utilizing dual reagents

A method for measuring luminescence of a biological sample, the method comprising:

• dispensing a biological sample into a well supported by a motorized sample carrier of a sample analyzing apparatus, the sample analyzing apparatus comprising a liquid dispensing system, the liquid dispensing system comprising a first injection needle, a second injection needle, a first reagent reservoir communicating with the first injection needle, and a second reagent reservoir communicating with the second injection needle;
• aligning the first injection needle with the well by moving the sample carrier, moving the first injection needle, or moving both the sample carrier and the first injection needle;
• dispensing a first reagent from the first injection needle into the well at a flow rate of 100 μL/sec or less by operating the liquid dispensing system, wherein the sample reacts with the first reagent to emit a first luminescent light;
• after a first delay period, measuring the first luminescent light at a luminescence detector over a first measurement period;
• dispensing a second reagent from the second injection needle into the well at a flow rate of 100 μL/sec or less by operating the liquid dispensing system while the well remains supported by the sample carrier, wherein the second reagent comprises a luminescent reagent effective for inducing emission of a second luminescent light, and a quenching reagent effective for suppressing emission of the first luminescent light, and wherein the sample reacts with the second reagent to emit the second luminescent light;
• after a second delay period, measuring the second luminescent light at the luminescence detector over a second measurement period;
• and
• agitating the well to enhance mixing of the second reagent with the sample by operating the sample carrier to agitate the well in an automated manner, wherein:
• the agitating is performed while the sample remains in the well and the well remains supported on the sample carrier;
• and
• the agitating is initiated at a time selected from the group consisting of:
• while dispensing the second reagent;
• during the second delay period;
• and while measuring the second luminescent light.