| Abstrakt: |
We present a new method to analyze the cytoplasmic contents of single cells in large cell populations. This new method consists of an array of microchambers in which individual cells are collected, enclosed, and lysed to create a reaction mixture of the cytoplasm with extracellular detection agents. This approach was tested for the analysis of red blood cells in 10,000 microchambers in parallel. Single cells were routinely collected in more than 60% of microchambers, the collected cells were robustly (up to 99%) lysed by electric fields, and the cytoplasm enclosed in each microchamber was analyzed with fluorescence microscopy. Using a heterogeneous cell mixture, we verified that the new method could distinguish individual cells by cytoplasmic composition and the analysis compared well with conventional flow-cytometric evaluation of mixed cell populations. In contrast to flow-cytometry, the new method monitored single cells over time, thus characterizing the distributions of caspase activities of 5000 individual cells. This approach should be interesting for a variety of applications that would benefit from the ability to measure the distribution of cytoplasmic compounds in complex cell populations, including hematology, oncology, and immunology. |
