Single-cell analysis of yeast, mammalian cells, and fungal spores with a microfluidic pressure-driven chip-based system.
BACKGROUND
Cytomics aims at understanding the function of cellular systems by analysis of single cells. Recently, there has been a growing interest in single cell measurements being performed in microfluidic systems. These systems promise to integrate staining, measurement, and analysis in a single system. One important aspect is the limitation of allowable cell sizes due to microfluidic channel dimensions. Here we want to demonstrate the broad applicability of microfluidic chip technology for the analysis of many different cell types.


METHODS
We have developed a microfluidic chip and measurement system that allows flow cytometric analysis of fluorescently stained cells from different organisms. In this setup, the cells are moved by pressure-driven flow inside a network of microfluidic channels and are analyzed individually by fluorescence detection.


RESULTS
We have successfully applied the system to develop a methodology to detect viable and dead cells in yeast cell populations. Also, we have measured short interfering RNA (siRNA) mediated silencing of protein expression in mammalian cells. In addition, we have characterized the infection state of Magnaportae grisea fungal spores.


CONCLUSIONS
Results obtained with the microfluidic system demonstrate a broad applicability of microfluidic flow cytometry to measurement of various cell types.
Palková
Zdena
Z
Department of Genetics and Microbiology, Charles University, Prague, Czech Republic.
Váchová
Libuse
L
Valer
Marc
M
Preckel
Tobias
T
D002460:Cell Line; D048429:Cell Size; D002470:Cell Survival / Q000502:physiology; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods*; D005456:Fluorescent Dyes; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D020082:Magnaporthe / Q000166:cytology / Q000254:growth & development; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D011312:Pressure; D012313:RNA / Q000032:analysis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000166:cytology; D013172:Spores, Fungal / Q000145:classification / Q000254:growth & development
D005456:Fluorescent Dyes; D012313:RNA