Single-Cell Multimodal Analytical Approach by Integrating Raman Optical Tweezers and RNA Sequencing

Abstract

Single-cell analysis has become a state-of-art approach to heterogeneity profiling in tumor cells. Herein, we realize a kind of single-cell multimodal analytical approach by combining single-cell RNA sequencing (scRNA-seq) with Raman optical tweezers (ROT), a label-free single-cell identification and isolation technique, and apply it to investigate drug sensitivity. The drug sensitivity of human BGC823 gastric cancer cells toward different drugs, paclitaxel and sodium dichloroacetate, was distinguished in the conjoint analytical way including morphology monitoring, Raman identification, and transcriptomic profiling. Each individual BGC823 cancer cell was measured by Raman spectroscopy, then nondestructively isolated out by ROT, and finally RNA-sequenced. Our results demonstrate each analytical mode can reflect cell response to the drugs from different perspectives and is consistent and complementary with each other. Therefore, we believe the multimodal analytical approach offers an access to comprehensive characterizations of the unicellular complexity, which especially makes sense for studying tumor heterogeneity or a desired special cell from a mixture cell sample such as whole blood.

Characterization of Drug Resistance in Chronic Myeloid Leukemia Cells Based on Laser Tweezers Raman Spectroscopy

“…16 Furthermore, extensive studies demonstrated successful classification in cancerous cells based on LTRS where the signals were mainly originated from DNA-RNA and proteins. [17][18][19] These results showed that the LTRS system has great potential to be a novel, convenient, and powerful tool for cell sorting and intracellular dynamics monitoring. However, whether LTRS could accurately discriminate drug sensitive and resistant cells remain unknown.…”

SRS-FISH: High-Throughput Platform Linking Microbiome Function to Identity at the Single Cell Level

“…These methods can be combined with fluorescence in situ hybridization (FISH) using rRNA-targeted probes 13 , enabling a direct link between function and identity of the organisms. In addition, recently Raman-activated cell sorting has been developed, using either optical tweezers or cell ejection for downstream sequencing of the sorted cells [14][15][16] . While these approaches have expanded the possibilities for functional analyses of microbiome members 17 , all of the aforementioned methods suffer from extremely limited throughput.…”

Rapid and accurate identification of pathogenic bacteria at the single‐cell level using laser tweezers Raman spectroscopy and deep learning

“…Raman spectroscopy is a label-free, rapid, and highly sensitive analytical technology [4][5][6][7], and in recent years, it has been used to identify bacteria [6,[8][9][10]. In various Raman technologies, surface-enhanced Raman spectroscopy (SERS) is a commonly used tool for bacterial analysis, which has been widely used in bacterial cell cycle monitoring [11], drug resistance [12], and identification [4,8,[13][14][15].…”