Method and apparatus for preparing cells for microtome sectioning and archiving nucleic acids and proteins
| Title: | Method and apparatus for preparing cells for microtome sectioning and archiving nucleic acids and proteins |
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| Patent Number: | 8,969,075 |
| Publication Date: | March 03, 2015 |
| Appl. No: | 12/338108 |
| Application Filed: | December 18, 2008 |
| Abstract: | A method and apparatus for embedding cells that utilizes a flow-through embedding technique maximizes the efficiency of extractions and decreases time for embedding the cell fragments, minimizes cell loss, and automatically positions cell samples at the position in which a microtome blade will section them. The apparatus includes a cell flow pathway defined by an inflow tube for delivering cell fragments from a cell sample to a sample port. The sample port is in fluid communication with a tissue cassette having attached thereto a filter. The cell flow pathway is in communication with a reagent flow pathway for delivering the reagents through the sample port to the cassette. The apparatus is configured such that the application of pressure directs the cell fragments from the cell sample through the cell flow pathway, and effects delivery of the reagents through the reagent flow pathway. The apparatus produces an embedded cell block having concentrated cells near the plane of the block to be sectioned in a quick and efficient manner. |
| Inventors: | Fischer, Andrew H. (Stow, MA, US) |
| Assignees: | University of Massachusetts (Boston, MA, US) |
| Claim: | 1. A flow-through system for forming an embedded cell block, comprising: a support for supporting a filter and a tissue cassette; a dispenser that automatically delivers a cell sample toward the support so that at least a portion of the cell sample is captured by the filter when the tissue cassette is provided to the support; a heated reservoir for storing a melted wax embedding solution; a heater configured to heat the melted wax embedding solution within the heated reservoir; one or more flow lines constructed and arranged to deliver at least one of a first reagent, a second reagent, and the melted wax embedding solution to the tissue cassette; and a pressure source fluidly communicable with the tissue cassette, when the tissue cassette is provided to the support, to create pressure to: urge the cell sample that has been delivered by the dispenser against the filter to capture at least a portion of the cell sample that is otherwise physically unrestrained; and urge at least one of the first reagent, the second reagent, and/or the melted wax embedding solution that has been delivered by the one or more flow lines through the otherwise physically unrestrained portion of the captured cell sample and the filter. |
| Claim: | 2. The system of claim 1 , wherein the pressure source includes a vacuum pump that creates a negative pressure to urge the otherwise physically unrestrained portion of the captured cell sample against the filter. |
| Claim: | 3. The system of claim 1 , wherein the pressure source creates a positive pressure to urge the otherwise physically unrestrained portion of the captured cell sample against the filter. |
| Claim: | 4. The system of claim 1 , wherein the dispenser includes an inflow tube. |
| Claim: | 5. The system of claim 4 , wherein the dispenser delivers the melted wax embedding solution to the otherwise physically unrestrained portion of the captured cell sample and the filter. |
| Claim: | 6. The system of claim 1 , wherein the dispenser and the one or more flow lines are integrated together. |
| Claim: | 7. The system of claim 1 , in combination with the tissue cassette that includes the filter. |
| Claim: | 8. The combination of claim 7 , wherein the support is a thermally conductive filter support positioned adjacent to the filter. |
| Claim: | 9. The system of claim 8 , further comprising: a heater that is configured to provide heat to the thermally conductive filter support. |
| Claim: | 10. The system of claim 9 , wherein the heater is configured to contact the thermally conductive filter support when the tissue cassette is present at the mount. |
| Claim: | 11. The system of claim 8 , further comprising: a cooler that is configured to draw heat from the thermally conductive filter support. |
| Claim: | 12. The system of claim 1 , further comprising: a fluid level monitor adapted to monitor a level of reagent about the otherwise physically unrestrained portion of the captured cell sample. |
| Claim: | 13. The system of claim 12 , wherein the fluid level monitor includes a monitor that measures a level of fluid provided to the otherwise physically unrestrained portion of the captured cell sample and the filter. |
| Claim: | 14. The system of claim 12 , wherein the fluid level monitor includes a monitor that measures an amount of fluid that has passed through the filter. |
| Claim: | 15. A flow through system for forming an embedded cell block, comprising: a support for supporting a filter and a tissue cassette, wherein the support is a thermally conductive filter support; a heater that is configured to heat the thermally conductive filter support when the tissue cassette is provided to the thermally conductive filter support; a dispenser that automatically delivers a cell sample toward the thermally conductive filter support so that at least a portion of the cell sample is captured by the filter, when the tissue cassette is provided to the thermally conductive filter support; one or more flow lines constructed and arranged to deliver at least one of a first reagent, a second reagent, and an embedding solution to the tissue cassette; and a pressure source in fluid communication with the tissue cassette, when the tissue cassette is provided to the thermally conductive filter support, to create pressure to: urge the cell sample that has been delivered by the dispenser against the filter to capture at least a portion of the cell sample; and urge at least one of a first reagent, a second reagent, and/or an embedding solution that has been delivered by the one or more flow lines through the portion of the captured cell sample and the filter. |
| Claim: | 16. The system of claim 15 , wherein the pressure source includes a vacuum pump that creates a negative pressure to urge the portion of the captured cell sample against the filter. |
| Claim: | 17. The system of claim 15 , wherein the pressure source creates a positive pressure to urge the portion of the captured cell sample against the filter. |
| Claim: | 18. The system of claim 15 , wherein the dispenser includes an inflow tube. |
| Claim: | 19. The system of claim 18 , wherein the dispenser delivers the embedding solution to the portion of the captured cell sample and the filter. |
| Claim: | 20. The system of claim 15 , wherein the dispenser and the one or more flow lines are integrated together. |
| Claim: | 21. The system of claim 15 , further comprising: a reservoir for storing the embedding solution prior to the embedding solution being dispensed. |
| Claim: | 22. The system of claim 21 , further comprising: a heater configured to heat the embedding solution within the reservoir. |
| Claim: | 23. The system of claim 15 , in combination with the tissue cassette that includes the filter and the thermally conductive filter support. |
| Claim: | 24. The system of claim 23 , further comprising: a cooler that is configured to draw heat from the thermally conductive filter support of the cassette. |
| Claim: | 25. The system of claim 15 , wherein the heater is configured to contact the thermally conductive filter support when the tissue cassette is present at the thermally conductive filter support. |
| Claim: | 26. The system of claim 15 , further comprising: a fluid level monitor adapted to monitor a level of reagent about the otherwise physically unrestrained portion of the captured cell sample. |
| Claim: | 27. The system of claim 26 , wherein the fluid level monitor includes a monitor that measures an amount of fluid provided to the otherwise physically unrestrained portion of the captured cell sample and the filter. |
| Claim: | 28. The system of claim 26 , wherein the fluid level monitor includes a monitor that measures an amount of fluid that has passed through the filter. |
| Current U.S. Class: | 4353/081 |
| Patent References Cited: | 2923669 February 1960 Poitra 3227130 January 1966 Weiskoff 4141312 February 1979 Louder et al. 4554839 November 1985 Hewett et al. 4557903 December 1985 McCormick 4569647 February 1986 McCormick 4604964 August 1986 Gordon et al. 4678559 July 1987 Szabados 4801553 January 1989 Owen et al. 5061452 October 1991 Yamamoto et al. 5080869 January 1992 McCormick 5112488 May 1992 Lemonnier 5137710 August 1992 Smalley et al. 5185084 February 1993 Lapidus et al. 5234585 August 1993 Zuk, Jr. 5266495 November 1993 Lapidus 5269671 December 1993 McCormick 5312758 May 1994 Ahlqvist 5354370 October 1994 Schmehl 5424040 June 1995 Bjornsson 5427742 June 1995 Holland 5532168 July 1996 Marantz 5665398 September 1997 McCormick 5667985 September 1997 O'Leary et al. 5691633 November 1997 Liu et al. 5817032 October 1998 Williamson, IV et al. 5843700 December 1998 Kerrod et al. 5928934 July 1999 McCormick 6010909 January 2000 Lapidus 6017476 January 2000 Renshaw 6042874 March 2000 Visinoni et al. 6207408 March 2001 Essenfeld et al. 6225125 May 2001 Lapidus 6372512 April 2002 Kerschmann 6395234 May 2002 Hunnell et al. 6413767 July 2002 Izvoztchikov et al. 6883958 April 2005 Mayer 6899850 May 2005 Haywood et al. 6905494 June 2005 Yon et al. 6905594 June 2005 Ferguson 6913921 July 2005 Fischer 7005110 February 2006 Taft et al. 7147826 December 2006 Haywood et al. 7156814 January 2007 Williamson, IV et al. 7541161 June 2009 Fischer 7829021 November 2010 Hutchins et al. 2005/0247646 November 2005 Linder 2006/0088930 April 2006 Smith et al. 2008/0057573 March 2008 Hutchins et al. 2008/0081351 April 2008 Hutchins et al. 2008/0081363 April 2008 Hutchins et al. 29 28 790 February 1981 0 245 969 November 1987 2278441 November 1994 2000-146782 May 2000 2002-525568 August 2002 2002-318177 October 2002 WO 00/16065 March 2000 WO 00/19897 April 2000 WO0067639 November 2000 WO 2006/096581 September 2006 |
| Other References: | European Search Report mailed Aug. 18, 2010 in connection with Application No. 10169650.8. cited by applicant Diaz-Rosario L. et al., “Cell Block Preparation by Inverted Filter Sedimentation is Useful in the Differential Diagnosis of Atypical Glandular Cells of Undetermined Significance in ThinPrep Specimens,” CANCER Cytopathology, American Cancer Society, 2000, pp. 265-272. cited by applicant Fahey C. et al., “Collodion Bag: A Cell Block Technique for Enhanced Cell Collection,” Histology, Laboratory Medicine, vol. 74, No. 2, Feb. 1993, pp. 94-96. cited by applicant “Vitae: The Magazine of the University of Massachusetts Medical School”, Annual Report 2006, vol. 29, No. 2, cover page and pp. 18-22. cited by applicant Srinivasan, M. et al., “Effect of Fixatives and Tissue Processing on the Content and Integrity of Nucleic Acids,” Review, American Journal of Pathology, vol. 161, No. 6, Dec. 2002, pp. 1961-1971. cited by applicant Yang, G. et al., “Compact Cell Block, Use of Body Fluids, Fine Needle Aspirations and Endometrial Brush Biopsies,” Acta-Cytologica, 1997, pp. 703-706. cited by applicant Office Action for U.S. Appl. No. 11/839,531 mailed Sep. 22, 2009. cited by applicant Office Action for U.S. Appl. No. 11/839,533 mailed Sep. 22, 2009. cited by applicant Translation of JP 2000-146782, (May 26, 2000). cited by applicant Translation of Japanese Office Action from Japanese Application 2010-156635 dated Aug. 1, 2012. cited by applicant Translation of JP 2002-286601 (not dated). cited by applicant |
| Primary Examiner: | Beisner, William H |
| Attorney, Agent or Firm: | Wolf, Greenfield & Sacks, P.C. |
| Accession Number: | edspgr.08969075 |
| Database: | USPTO Patent Grants |
| Language: | English |
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