Ultrasound-mediated high-speed biological reaction and tissue processing

• US 6,291,180 B1
• Filed: 09/29/1999
• Issued: 09/18/2001
• Est. Priority Date: 09/29/1999
• Status: Expired due to Fees

• Current Assignee: American Registry LLC
• Original Assignee: American Registry LLC
• Inventors: Chu, Wei-Sing
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/407,964
• Time in Patent Office: 720 Days
• Field of Search: 427/2.13, 427/4, 435/1.1, 435/1.3, 435/40.5, 435/40.52, 435/325, 435/6, 422/20
• US Class Current: 435/6.11
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 1/38 Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 1/44 Sample treatment involving radiation, e.g. heat
• CPC Class Codes: G01N 2291/02475 Tissue characterisation
• CPC Class Codes: G01N 2291/02881 Temperature
• CPC Class Codes: G01N 29/223 Supports, positioning or alignment in fixed situation mounting transducers per se G10K11/004
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack

Abstract

Methods of fixing and processing tissue and samples on a membrane by using ultrasound radiation as a part of the method are presented. Ultrasound of a frequency in the range of 0.1-50 MHz is used and the sample or tissue receives 0.1-200 W/cm² of ultrasound intensity. The use of ultrasound allows much shorter times in the methods. Also presented are apparati comprising transducers of one or of multiple heads for producing the ultrasound radiation and further comprising a central processing unit and optionally comprising one or more sensors. Sensors can include those to measure and monitor ultrasound and temperature. This monitoring system allows one to achieve accurate and optimum tissue fixation and processing without overfixation and tissue damage. The system also allows the performance of antigen-antibody reactions or nucleic acid hybridizations to be completed in a very short time while being highly specific and with a very low or no background.

Claims

1. A method for fixing or processing a tissue sample comprising exposing said tissue sample to ultrasound of a frequency of at least 100 Khz wherein said ultrasound is produced by an ultrasound transducer.

2. The method of claim 1 wherein only a single piece of said tissue sample is placed into said transducer.

3. The method of claim 1 wherein said frequency is in the range of 100 KHz to 50 MHz and wherein said frequency is a single frequency or a wideband frequency.

4. The method of claim 1 wherein two or more ultrasound transducers are used to produce ultrasound.

5. The method of claim 1 wherein one or more ultrasound transducers are used to produce an ultrasound field such that at least a portion of said tissue sample receives ultrasound of a uniform frequency and a uniform intensity.

6. The method of claim 1 wherein said transducer comprises only one head.

7. The method of claim 6 wherein said head is capable of emitting a wideband frequency.

8. The method of claim 6 wherein said head is capable of emitting a single frequency or a wideband frequency.

9. The method of claim 1 wherein said transducer comprises multiple heads.

10. The method of claim 9 wherein one or more of said multiple heads are capable of emitting a wideband frequency.

11. The method of claim 9 wherein one head on a single transducer produces a frequency different from a frequency produced by a second head on said single transducer.

12. The method of claim 9 wherein one head on a single transducer produces a range of frequencies and a range of intensities different from a range of frequencies and a range of intensities produced by a second head on said single transducer.

13. The method of claim 4 wherein each of said transducers produces an ultrasound frequency different from an ultrasound frequency produced by at least one other transducer.

14. The method of claim 4 wherein each of said transducers produces a range of ultrasound frequencies and a range of ultrasound intensities different from a range of ultrasound frequencies and a range of ultrasound intensities produced by at least one other transducer.

15. The method of claim 13 wherein a range of frequencies is applied to said tissue sample.

16. The method of claim 4 wherein said transducers are arranged around said tissue sample in a two-dimensional arrangement.

17. The method of claim 4 wherein said transducers are arranged around said tissue sample in a three-dimensional arrangement.

18. The method of claim 1 wherein said tissue sample is rotated.

19. The method of claim 1 wherein said transducer revolves around said tissue sample.

20. The method of claim 1 wherein said ultrasound is produced as a continuous signal.

21. The method of claim 20 wherein said ultrasound is a single frequency in the range of 0.1-50 MHz.

22. The method of claim 20 wherein said ultrasound is a wideband frequency in the range of 0.1-50 MHz.

23. The method of claim 1 wherein said ultrasound is produced in pulses.

24. The method of claim 23 wherein said ultrasound is a single frequency in the range of 0.1-50 MHz.

25. The method of claim 23 wherein said ultrasound is produced as a wideband frequency in the range of 0.1-50 MHz.

26. The method of claim 23 wherein said pulses vary in frequency in the range of 0.1-50 MHz.

27. The method of claim 23 wherein said pulses vary in intensity.

28. The method of claim 23 wherein said ultrasound is produced as a continuous signal.

29. The method of claim 28 wherein over time said signal varies in frequency in the range of 0.1-50 MHz.

30. The method of claim 28 wherein over time said signal varies in intensity.

31. The method of claim 1 wherein said tissue sample receives ultrasound of a power of at least 5 W/cm².

32. The method of claim 1 wherein said tissue sample receives ultrasound with a power in the range of 5-150 W/cm².

33. The method of claim 1 further comprising using one or more sensors to detect one or more parameters of reflected ultrasound wherein said parameters are selected from the group consisting of intensity and frequency.

34. The method of claim 33 wherein more than one type of sensor is used.

35. The method of claim 34 comprising an ultrasound sensor and a sensor to measure temperature.

36. The method of claim 33 further comprising a central processing unit to monitor the sensor readings.

37. The method of claim 36 wherein said central processing unit controls said ultrasound generator.