Quantitative Intracellular Molecular Profiling Using a One-Dimensional Flow System

Abstract

We report on the development of one-dimensional microfluidic bead arrays for rapid and quantitative molecular profiling of human cancer cells. This new bioanalytical platform integrates the rapid binding kinetics of suspension bead carriers, the multiplexing and encoding capabilities of gene/protein chips, and the liquid handling advantages of microfluidic devices. Using antibody-conjugated beads in a two-site "sandwich" format, we demonstrate that the proteomic contents of as few as 56 human lung epithelial cancer cells can be determined with high sensitivity and specificity. The results indicate that each cell contains approximately 6 x 10(5) copies of the tumor suppressor protein P53. We have further examined the expression changes of P53, c-Myc, and beta-Actin as a function of anticancer drug treatment and have validated these changes by using Western blotting. This ability to quantitatively analyze normal and diseased cells raises new possibilities in studying cancer heterogeneity and circulating tumor cells.

Detection of Circulating Tumor Cells in Peripheral Blood of Patients with Renal Cell Carcinoma Correlates with Prognosis

“…Briefly, there are two main methods: (a) immunologic assays using immunocytochemistry and (b) PCR-based assays, which investigate tissue-specific transcripts. Both methods include an enrichment of circulating tumor cells by immunomagnetic techniques (6), flow cytometry (7), Ficoll density centrifugation (8), or microchips (9,10). Only the first method allows for the isolation and evaluation of circulating tumor cells by a pathologist.…”

HER2 status predicts the presence of circulating tumor cells in patients with operable breast cancer

“…Potential issues when interpreting the CTC literature include differing assays used to detect CTCs, potentially inadequate follow-up duration, small numbers of patients studied and in some cases analyzing operable and metastatic patients together rather than separately. Methods to isolate CTCs vary considerably and include immunomagnetic enrichment [11], flow-cytometry [12], Ficoll density centrifugation [4] and microfluidics chips [13]. Additional well designed studies are required to determine the significance of CTCs in operable breast cancer patients.…”

Original article. Potential risk of dihydroergotamine causing medicationoveruse headache: preclinical evidence

“…Second, we attempted to quantitate total c-Fos expression in the TNC by western blotting analysis. Western blotting offers advantages of sensitivity, specificity, and protein size determination Dihydroergotamine medication-overuse headache [12]. It is undeniable that low expression of c-Fos by weakly immunoreactive nuclei residing in the dorsal horn was also included when we have evaluated the change of c-Fos expression associated with chronic DHE treatment compared with the control groups.…”