cell enrichment extraction (cee™) microchannels  (Biocept Inc)

Journal: Journal of Oncology

Article Title: Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood

doi: 10.1155/2011/252361

Figure Lengend Snippet: Diagram of the CEE microchannel. (a) Top view of the channel showing the inlet where sample is loaded and the outlet that is attached to a syringe pump to draw sample through the channel. (b) Bottom view shows the area where 9,000 posts are located in the silicone block and the channel sealed with the bottom cover slip. The total volume of the microchannel is 24 μ L. A standard microscope slide is added for stability during handling but is removed to visualize cells. The microchannel is inverted on a microscope and the captured cells viewed through the coverslip.

Article Snippet: The final pellet was suspended in 1 mL PBS/BSA/EDTA and subjected to capture and staining on the Cell Enrichment and Extraction (CEE) microchannel (manufactured at Biocept, Inc., San Diego, Calif.).

Techniques: Blocking Assay, Microscopy

Journal: Journal of Oncology

Article Title: Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood

doi: 10.1155/2011/252361

Figure Lengend Snippet: Immunofluorescent staining. (a) A LnCAP cell spiked into blood and captured on the microchannel, stained for CK (green) and also nuclear stained with DAPI (blue). A small WBC is seen with only the nucleus stained blue. (b) A cluster of CTCs from a clinical lung cancer sample captured on the microchannel that are stained for CK. These cells were CD45-negative and DAPI-positive (not shown). (c) A cluster of cells from lung cancer showing triple staining with CK (green), CD45 (red), and DAPI (blue). Three CK-positive CTCs are shown with 2 smaller WBCs stained positive for CD45.

Article Snippet: The final pellet was suspended in 1 mL PBS/BSA/EDTA and subjected to capture and staining on the Cell Enrichment and Extraction (CEE) microchannel (manufactured at Biocept, Inc., San Diego, Calif.).

Techniques: Staining

Journal: Journal of Oncology

Article Title: Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood

doi: 10.1155/2011/252361

Figure Lengend Snippet: The use of CEE-Enhanced to improve detection of cells on the microchannel. (a) A clinical breast cancer CTC stained for CK and nuclear-stained with DAPI. This cell is weakly CK positive. (b) The same cell after subsequent stain with CE labeled with the same AlexaFLuor-488 fluorophore in order to enhance the stain intensity. (c) SKOV cell spiked into blood and recovered on the microchannel using an antibody mixture (see ). Cells on the channels were stained with CE-488 and DAPI. The four WBCs stained blue for DAPI only, while the SKOV (attached to post) can be detected only with CE (green). This higher contrast image shows the outline of the posts in the microchannel. Together these images show that CE can be used to augment weakly staining CK cells or can be used to detect cells without CK stain.

Article Snippet: The final pellet was suspended in 1 mL PBS/BSA/EDTA and subjected to capture and staining on the Cell Enrichment and Extraction (CEE) microchannel (manufactured at Biocept, Inc., San Diego, Calif.).

Techniques: Staining, Labeling

Journal: Journal of Oncology

Article Title: Detection of EpCAM-Negative and Cytokeratin-Negative Circulating Tumor Cells in Peripheral Blood

doi: 10.1155/2011/252361

Figure Lengend Snippet: Costaining clinical lung cancer CTCs with anti-CK and CEE-Enhanced. (a–c) A single CTC on the microchannel stained with anti-CK (a), CE-AlexaFluor-546 (orange, (b)), and (c), a composite image. (d–f) shows the same order of staining but with 2 attached CTCs. This demonstrates the costaining of the internal CK antigen and the cell surface antigens with CE.

Article Snippet: The final pellet was suspended in 1 mL PBS/BSA/EDTA and subjected to capture and staining on the Cell Enrichment and Extraction (CEE) microchannel (manufactured at Biocept, Inc., San Diego, Calif.).

Techniques: Staining