ja08拉曼癌细胞支持材料.pdfVIP

Supporting Information Surface Enhanced Raman Scattering Tags for Rapid and Homogeneous Detection of Circulating Tumor Cells in Presence of Human Whole Blood Michael Y. Sha*, Hongxia Xu, Michael J. Natan and Remy Cromer Oxonica Inc. 325 E.. Middlefield Rd, Mountain View, CA 94043. * mike_sha168@ MATERIALS AND METHODS Cell line and cell culture SKBR3 (HTB-30) cell line was purchased from ATCC. The vial was thawed with gentle agitation in a 37°C water bath. The vial contents were transferred to a centrifuge tube containing 9.0 ml complete culture medium and spun at approximately 125 x g for 5 to 7 minutes. The cell pellet was re-suspended with 9 ml of McCoy5 with10% FBS and dispensed into a 25 cm2 or a 75 cm2 culture flask. Prior to the addition of the vial contents, the culture vessel containing the complete growth medium has been placed into the incubator for at least 15 minutes to allow the medium maintaining its normal pH (7.0 to 7.6). The cell culture was incubated in an incubator at 37°C with 5% CO2 in air atmosphere. Magnetic bead conjugated with EpCAM antibodies 4.5 um Dynalbead (Collection Epithelial Enrich, Cat number 162-03) was purchased from Invitrogen Inc. This magnetic bead is already conjugated with EpCAM antibody. Nanoplex TM biotag synthesis and functionalization TM 1 The synthesis of the Nanoplex Biotags has been described by Mulvaney et al . Briefly, a 50 nm spherical Au colloid is coated first with a layer containing both a glass precursor silane and the Raman label molecule. A thin (a few nanometers) silica coating