Analytical Techniques for Single-Cell Studies in Microbiology

Author(s):  
Evgeny Puchkov
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chen Chen ◽  
Zhilun Zhao ◽  
Naixin Qian ◽  
Shixuan Wei ◽  
Fanghao Hu ◽  
...  

AbstractSingle-cell multiparameter measurement has been increasingly recognized as a key technology toward systematic understandings of complex molecular and cellular functions in biological systems. Despite extensive efforts in analytical techniques, it is still generally challenging for existing methods to decipher a large number of phenotypes in a single living cell. Herein we devise a multiplexed Raman probe panel with sharp and mutually resolvable Raman peaks to simultaneously quantify cell surface proteins, endocytosis activities, and metabolic dynamics of an individual live cell. When coupling it to whole-cell spontaneous Raman micro-spectroscopy, we demonstrate the utility of this technique in 14-plexed live-cell profiling and phenotyping under various drug perturbations. In particular, single-cell multiparameter measurement enables powerful clustering, correlation, and network analysis with biological insights. This profiling platform is compatible with live-cell cytometry, of low instrument complexity and capable of highly multiplexed measurement in a robust and straightforward manner, thereby contributing a valuable tool for both basic single-cell biology and translation applications such as high-content cell sorting and drug discovery.


2010 ◽  
Vol 398 (6) ◽  
pp. 2493-2504 ◽  
Author(s):  
Andrea Amantonico ◽  
Pawel L. Urban ◽  
Renato Zenobi

2021 ◽  
Author(s):  
Chen Chen ◽  
Zhilun Zhao ◽  
Naixin Qian ◽  
Shixuan Wei ◽  
Fanghao hu ◽  
...  

Abstract Single-cell multiparameter measurement has been increasingly recognized as a key technology toward systematic understanding of complex molecular and cellular functions in biological systems. Despite extensive efforts in analytical techniques, it is still generally challenging for existing methods to decipher a large number of phenotypes in a single living cell. Herein we devise a super-multiplexed Raman probe panel with sharp and mutually resolvable Raman peaks to simultaneously quantify cell surface proteins, endocytosis activities, and metabolic dynamics of an individual live cell. When coupled it to whole-cell spontaneous Raman micro-spectroscopy, we demonstrate the utility of this technique in 14-plexed live-cell profiling and phenotyping under various drug perturbations. In particular, single-cell multiparameter measurement enables powerful clustering, correlation, and network analysis with biological insights. Being the highest Raman-based multiplexing technology of biological targets so far, this profiling platform is compatible with live cell cytometry, of low instrument complexity and capable of highly multiplexed measurement in a robust and straightforward manner, thereby contributing a valuable tool for both basic single-cell biology and translation applications such as high-content cell sorting and drug discovery.


The Analyst ◽  
2015 ◽  
Vol 140 (11) ◽  
pp. 3687-3695 ◽  
Author(s):  
Christian Amatore ◽  
Jérôme Delacotte ◽  
Manon Guille-Collignon ◽  
Frédéric Lemaître

Among all the analytical techniques capable of monitoring exocytosis in real time at the single cell level, electrochemistry (particularly amperometry at a constant potential) using ultramicroelectrodes has been demonstrated to be an important and convenient tool for more than two decades.


Author(s):  
R. E. Herfert

Studies of the nature of a surface, either metallic or nonmetallic, in the past, have been limited to the instrumentation available for these measurements. In the past, optical microscopy, replica transmission electron microscopy, electron or X-ray diffraction and optical or X-ray spectroscopy have provided the means of surface characterization. Actually, some of these techniques are not purely surface; the depth of penetration may be a few thousands of an inch. Within the last five years, instrumentation has been made available which now makes it practical for use to study the outer few 100A of layers and characterize it completely from a chemical, physical, and crystallographic standpoint. The scanning electron microscope (SEM) provides a means of viewing the surface of a material in situ to magnifications as high as 250,000X.


Author(s):  
R.W. Horne

The technique of surrounding virus particles with a neutralised electron dense stain was described at the Fourth International Congress on Electron Microscopy, Berlin 1958 (see Home & Brenner, 1960, p. 625). For many years the negative staining technique in one form or another, has been applied to a wide range of biological materials. However, the full potential of the method has only recently been explored following the development and applications of optical diffraction and computer image analytical techniques to electron micrographs (cf. De Hosier & Klug, 1968; Markham 1968; Crowther et al., 1970; Home & Markham, 1973; Klug & Berger, 1974; Crowther & Klug, 1975). These image processing procedures have allowed a more precise and quantitative approach to be made concerning the interpretation, measurement and reconstruction of repeating features in certain biological systems.


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