2026-10-14T02:33:51Z https://www.scidb.cn/oai

10.57760/sciencedb.28654 2026-05-07T11:04:10Z

2026-05-07 Detection of bacteria-residing circulating tumor cells (CTCs) using DARMR doi:10.57760/sciencedb.28654 en Fresh blood samples (3–5 mL) from cancer patients were collected into EDTA-coated tubes and immediately labeled with 10 μM DARMR, incubated at 37°C for 0.5 h. Red blood cells were lysed twice by two volumes of ACK lysis buffer (C3702-500mL, Beyotime biotechnology) on ice for 5 min. The pellet was resuspended in PBS, and CTC isolation was performed using a size-dictated immunocapture chip (SDI-Chip) as described previously.The SDI-Chip was fabricated using PDMS through soft lithography, with SU8-3050 photoresist (Alfa Chemistry #SU8-3050) for micropillar mold preparation. Surface modification involved 4% MPTS (3-mercaptopropyl trimethoxysilane; Sigma-Aldrich #175617-100G), 0.01 μmol/mL GMBS (N-γ-maleimidobutyryl-oxysuccinimide ester; Sigma-Aldrich #442630-100MG), and 20 μg/mL biotinylated anti-EpCAM antibody immobilized via streptavidin (Sigma-Aldrich #S4762). Nonspecific binding was blocked with 3% BSA /0.05% Tween 20 in PBS.Samples were introduced into the chip at 1 mL/h using a syringe pump, and captured cells were released with 0.25% trypsin at 37°C for 10 min. Viability was assessed by Calcein-AM/PI (Solarbio #CA1630), and immunostaining used Mouse anti-human Cytokeratin Antibody anti-human FITC (1:200, Miltenyi #130-118-964), Rabbit anti-human CD68 Alexa Fluor® 647 (1:200, Biolegend #333820), mouse anti-human CD45 APC (1:200, BD Biosciences #555485), and DAPI (1:1000, ThermoFisher Scientific #D1306) for nuclear counters taining. Images were acquired on a Zeiss LSM 900 confocal microscope (Carl Zeiss, Germany) equipped with Airyscan 2 super-resolution detector using a 10×/0.45 NA Plan-Apochromat objective. peripheral blood ; CTC; bacteria Yuezhou Wang Zile Jiang Jinshan Gao Yiqun Huang Jinsong Ma Kai Zhang Huimin Tang Guimei Wang Jie Wang Fei Ma Zhicheng Yang Anqi Ding Hu Zhou Xiang Zhi Baoyue Liang Xuehui Hong Xianming Deng EMBARGO https://creativecommons.org/licenses/by-nc-nd/4.0/ dataset Science Data Bank