Device and method for in vivo flow cytometry using the detection of photoacoustic waves

• US 9,217,703 B2
• Filed: 10/26/2012
• Issued: 12/22/2015
• Est. Priority Date: 12/13/2007
• Status: Active Grant

• Current Assignee: Bioventures, LLC
• Original Assignee: The Board of Trustees of the University of Arkansas (University of Arkansas System)
• Inventors: Zharov, Vladimir Pavlovich
• Primary Examiner(s): Le, Long V
• Assistant Examiner(s): IMPINK, BRADLEY GERALD
• Application Number:
  US13/661,551
• Publication Number:
  US 20130060122A1
• Time in Patent Office: 1,152 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61B 18/20 using laser
• CPC Class Codes: A61B 2018/00642 with feedback, i.e. closed loop control
• CPC Class Codes: A61B 2018/20351 Scanning mechanisms
• CPC Class Codes: A61B 2018/20357 by movable optical fibre end
• CPC Class Codes: A61B 2018/20361 with redirecting based on sensed condition, e.g. tissue analysis or tissue movement
• CPC Class Codes: A61B 2018/207 mixing two wavelengths
• CPC Class Codes: A61B 5/0059 using light, e.g. diagnosis by transillumination, diascopy, fluorescence A61B5/0093, A61B5/1455, A61B5/02416 and A61B5/0261 take precedence
• CPC Class Codes: A61B 5/0095 by applying light and detecting acoustic waves, i.e. photoacoustic measurements
• CPC Class Codes: A61B 5/02007 Evaluating blood vessel condition, e.g. elasticity, compliance
• CPC Class Codes: A61B 5/412 Detecting or monitoring sepsis
• CPC Class Codes: A61B 5/415 the glands, e.g. tonsils, adenoids or thymus
• CPC Class Codes: A61B 5/416 the spleen
• CPC Class Codes: A61B 5/418 lymph vessels, ducts or nodes
• CPC Class Codes: A61B 5/7278 Artificial waveform generation or derivation, e.g. synthesising signals from measured signals A61B5/327 takes precedence
• CPC Class Codes: A61B 8/08 Detecting organic movements or changes, e.g. tumours, cysts, swellings A61B8/02 - A61B8/06 take precedence
• CPC Class Codes: A61K 49/22 Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
• CPC Class Codes: G01N 15/1425 using an analyser being characterised by its control arrangement
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 21/1702 : with opto-acoustic detection, e.g. for gases or analysing solids
• CPC Class Codes: G01N 21/39 : using tunable lasers
• CPC Class Codes: G01N 2201/0697 : Pulsed lasers

Abstract

A photoacoustic flow cytometry (PAFC) device for the in vivo detection of cells circulating in blood or lymphatic vessels is described. Ultrasound transducers attached to the skin of an organism detect the photoacoustic ultrasound waves emitted by target objects in response to their illumination by at least one pulse of laser energy delivered using at least one wavelength. The wavelengths of the laser light pulse may be varied to optimize the absorption of the laser energy by the target object. Target objects detected by the device may be unlabelled biological cells or cell products, contrast agents, or biological cells labeled with one or more contrast agents.

Claims

1. A method for detecting at least one type of moving target object within a circulatory vessel of a living organism, the method comprising:

a. pulsing a target object moving through a vessel with at least two laser pulses at a region of detection, wherein the at least two laser pulses comprise a first laser pulse with a first wavelength and a second laser pulse with a second wavelength different from the first wavelength and the second laser pulse is separated from the first laser pulse by a time delay ranging between about 0.1 μ

s and about 100 μ

s, and wherein each laser pulse is delivered as a beam with a maximum cross-sectional dimension of about a vessel diameter;

b. detecting at least two photoacoustic pulses emitted by the target object in response to the at least two laser pulses; and

,

c. analyzing the at least two photoacoustic pulses to determine at least one characteristic of the target object.

2. The method of claim 1, wherein each of the at least two laser pulses is delivered at a pulse width ranging between about 0.1 ps and about 1000 ns, a pulse repeat rate ranging between about 1000 Hz and about 500,000 Hz, and a pulse energy fluence ranging between about 0.1 mJ/cm² and about 1000 J/cm², wherein each laser pulse beam has a maximum dimension ranging between about 1 μ

m and about 150 μ

m, and wherein the first wavelength is between about 400 and about 2500 nm and the second wavelength is at a different wavelength between about 400 and about 2500 nm.

3. The method of claim 1, wherein the at least two photoacoustic pulses are detected with at least one ultrasound transducer with a sample rate ranging between about 10 kHz and about 100 MHz.