Patent SG-11201807053T-A (SG)
Title: Compositions and methods for identifying rare cells
Inventor: BHAKDI SEBASTIAN
IPC: G01N21/75
Filed: 2017-03-06
Abstract: ocentrifugea cell onto a slide T — — optionally Apply a first fixing buffer to the cell, the first fixing buffer including a hydrophilic polymer diluted in alcohol AppIya second fixing buffer to the cell, the second fixing buffer including a hydrophilic polymer, a detergent, and a fixative Silo S S13O W O 20 17 / 155 869 Al S150 buffer including a hydrophilic polymer, detergent, and hydrolyzed collagen ... ...... t optionally stain the cell Apply a blocking buffer to the cell, the blocking F10.1 (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/155869 Al 14 September 2017 (14.09.2017) WIPO I PCT 111111111111110111011111111111010111110 01111111111111110111111110111111111110111111 (72) Inventor: BHAKDI, Sebastian, Chalirit, Pun- yaratabandhu; Lahnbergstrasse 17a, 35043 Marburg (DE). (74) Agent: SLOAT, Ashley; Aurora Consulting LLC, 1482 Woods And Waters Trl., Traverse City, MI 49696 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, (51) International Patent Classification: C01F 7/04 (2006.01) Cl 2N 11/08 (2006.01) CO1G 37/10 (2006.01) C12Q 1/68 (2006.01) Cl 1D 3/12 (2006.01) G01N 21/75 (2006.01) C11D 9/22 (2006.01) (21) International Application Number: PCT/US2017/020905 (22) International Filing Date: 6 March 2017 (06.03.2017) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 62/304,452 7 March 2016 (07.03.2016) US 62/313,366 25 March 2016 (25.03.2016) US 62/313,250 25 March 2016 (25.03.2016) US 62/430,542 6 December 2016 (06.12.2016) US (71) Applicant: X-ZELL INC. [US/US]; 640 West California Ave., Suite 210, Sunnyvale, CA 94086 (US). [Continued on next page] (54) Title: COMPOSITIONS AND METHODS FOR IDENTIFYING RARE CELLS (57) : Disclosed herein are compositions and methods of fix- 100 ing and staining rare cells. Farther, disclosed herein are methods of identifying circulating tumor cells (CTC). In some embodiments, the method includes: imaging a cell sample to identity a cell of interest; determining a first pixel intensity of a stained nuclear area; determin- ing a second pixel intensity of a background area; calculating a ploidy status of the cell of interest by subtracting the second pixel intensity from the first pixel intensify; and determining whether the cell of in- terest is a CTC based on the ploidy status. The method may be com- puter implemented, such, that the method uses a machine learning al- gorithm to identify a feature; process the feature to extract a parameter of interest; analyze the parameter of interest; and when the parameter of interest is greater than or less than a predetermined threshold, classi- fy the cell of interest as a CTC. 5140 optionally WO 2017/155869 Al MIDEDIMOMOIDEIR010131MMEIMMOVOIMIE TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Published: with international search report (Art. 21(3)) before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h))