Flow Measurement and Control for Improved Quantification of Particles in Flow Cytometry
AssigneeIndevr, Inc.
InventorsKathy Rowlen, Matthew Ferris
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods that enable improved accuracy for quantitative particle counting in a flowing liquid stream. The methods of the present invention utilize the real-time measurement of flow rates and flow rate control through feedback mechanisms to improve quantification, and this improved quantification translates to more accurate particle counting. In certain embodiments, particles being counted are biological particles in a liquid sample, such as viruses.
Claims19
A system to determine a number of particles in a fluid volume in real time or near real time, comprising:
- at least one compressed fluid supply;
- a sample reservoir containing sample fluid, the at least one compressed fluid supply being in fluid communication with the sample reservoir;
- a sample pressure regulator in fluid communication with the at least one compressed fluid supply and the sample reservoir to regulate a pressure of the sample reservoir;
- a sheath reservoir containing sheath fluid, the at least one compressed fluid supply being in fluid communication with the sheath reservoir;
- a sheath pressure regulator in fluid communication with the at least one compressed fluid supply and the sheath reservoir to regulate a pressure of the sheath reservoir;
- a flow cell, the flow cell being in fluid communication with both the sample reservoir and the sheath reservoir;
- a sample flow sensor in fluid communication with the sample reservoir and the flow cell, the sample flow sensor coupled to the sample pressure regulator to alter the pressure of the sample reservoir to maintain a substantially constant sample flow rate;
- an excitation and detection system coupled to the flow cell to count the number of particles flowing through the flow cell, wherein the total number of particles in the volume of fluid is provided in real or near real time.
A method of using controlled flow of a sample fluid to allow for quantification of particles in the sample fluid stream in real or near real time, the method comprising:
- providing a sample fluid have a plurality of particles in a reservoir in fluid communication with a flow cell;
- pressurizing the sample fluid reservoir to cause the sample fluid to flow from the sample fluid reservoir to the flow cell;
- measuring a flow parameter of the sample fluid flowing between the sample fluid reservoir and the flow cell;
- merging a sheath fluid flow with the sample fluid flow prior to entering the flow cell to focus the sample fluid flow in the flow cell;
- controlling the pressure of the sample fluid reservoir using the measured flow parameter to maintain the sample fluid flow at a substantially constant flow rate;
- counting a number of particles passing through the flow cell;
- and
- providing the number of particles in a fluid volume in real or near real time.
A system to determine a number of particles in a fluid volume in real time or near real time, comprising:
- at least one compressed fluid supply;
- a sample reservoir containing sample fluid, the at least one compressed fluid supply being in fluid communication with the sample reservoir to pressurize the sample reservoir;
- means to control the pressurization of the sample reservoir;
- a sheath reservoir containing sheath fluid, the at least one compressed fluid supply being in fluid communication with the sheath reservoir to pressurize the sheath reservoir;
- a flow cell, the flow cell being in fluid communication with both the sample reservoir and the sheath reservoir;
- means to control the flow rate of the sample fluid flow between the sample fluid reservoir and the flow cell at a substantially constant flow rate;
- and
- an excitation and detection system optically coupled to the flow cell to count the number of particles flowing through the flow cell, wherein the total number of particles in the volume of fluid is provided in real or near real time.
Dependencies are inferred from the claim text.