Activating mutants in rat sarcoma （RAS） and B-rapid accelerated fibrosarcoma （BRAF） are found in at least a third of cases of human tumors and melanoma; hence, numerous therapeutic treatments target this pathway. In this letter, we study the adhesion force of RAS-coated beads with BRAF-coated beads, BRAF （A246P） mutant-coated beads, and GST-coated beads using optical tweezers. One full and two fractional RAS BRAF specific binding modes are identified using the rupture force distribution. The koff（0） of the full binding mode in RAS BRAF is 3.71×10^-4/s and 1.16×10^-4s^-1 in RAS BRAF （A246P）, whereas the Xb is around 3×10^-10 m in both groups.
The ability to discriminate between single cells in a label-free and noninvasive fashion is important for the classification of cells, and for the identification of similar cells from different origins. In this paper, we present the Raman spectroscopy-based identifi- cation of different types of single cells in aqueous media, and discrimination between the same types of cells from different donors using a novel Laser Tweezers Raman Spectroscopy (LTRS) technique, which combines laser trapping and micro-Raman spectroscopy. First, we measured the spectra of individual living human erythrocytes, i.e. red blood cells, and leucocytes (U937 cancer cells). High-quality Raman spectra with low fluorescence were obtained using a home-LTRS apparatus and 20 cells were measured for each cell type. The smoothing, baseline subtraction, and normalization of the data were followed by a principal components analysis (PCA). The PCA loading plots showed that the two different types of cells could be completely separated based only on the first component (PC1) (i.e. the peaks at 1300 cm1 ); the discrimination accuracy could therefore reach 100%. More than 50 spectra were taken for each erythrocyte obtained from the four healthy volunteers. The average discrimination accuracy was 84.5% for two random individuals taken from the four volunteers, according to the first and second PCs. This work demonstrates that LTRS is a powerful tool for the accurate identification and discrimination of single cells, and it has the potential to be applied for the highly sensitive identification of cells in clinical diagnosis and medical jurisprudence.
We investigate the activation of living monocytic U937 cells induced by interleukin-6 （IL-6） at the single cell level. We employ home-built Raman tweezers to measure the Raman spectra of living U937 cells with and without IL-6 at the single cell level. Raman peaks of amide III, amide I, DNA backbone, as well as guanine and adenine in U937 cells, change at 1312, 1652, 1090, and 1576 cm ？1 , respectively, shortly after IL-6 is added in the medium. The change is a dynamic temporal process. In the activation process of U937 cells induced by IL-6, the protein signals recover in 20 min, while the nucleic acid signals continue to increase for 20 min. The results reveal that the biochemical cascades of activation in signal transduction induced by IL-6 can be investigated in situ at the single cell level.
