Intracellular proteins imaging

Imaging of Intracellular Proteins Imaging of Transmembrane Proteins... 

One major advantage of DHLA modification is that the diameter of the QD remains as small as possible, while still creating a water-soluble particle. QDs of <10nm diameter have been created using this process and were successfully used to image intracellular proteins (Jaiswal et al., 2003). 

Key words Covalent labeling, E3 ubiquitin Hgase, Fluorescence, Imaging, Intracellular, Protein degradation signal, Protein stability, Pulse chase... 

Asymmetric localization of intracellular proteins and signals directs movement dimng axon guidance, endothelial cell invasion, and immune cell migration. In these processes, cell movement is guided by external chemical cues in a process known as chemotaxis. In particular, leukocyte migration in the innate immune system has been studied in the human neutrophil-like cell line (HL-60). Here, we describe the maintenance and transfection of HL-60 cells and explain how to analyze their behavior with two standard chemotactic assays. Finally, we demonstrate how to fix and stain the actin cytoskeleton of polarized cells for fluorescent microscopy imaging. 

Stehbens S, Pemble H, Mmrow L et al (2012) Imaging intracellular protein dynamics by spinning disk confocal microscopy. Methods Enzymol 504 293-313... 

Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess FIF (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313 1642-1645... 

Bastiaens, P. I. and Jovin, T. M. (1996). Microspectroscopic imaging tracks the intracellular processing of a signal transduction protein fluorescent-labeled protein kinase C beta I. Proc. Natl. Acad. Sci. USA 93, 8407-12. 

Various optical detection methods have been used to measure pH in vivo. Fluorescence ratio imaging microscopy using an inverted microscope was used to determine intracellular pH in tumor cells [5], NMR spectroscopy was used to continuously monitor temperature-induced pH changes in fish to study the role of intracellular pH in the maintenance of protein function [27], Additionally, NMR spectroscopy was used to map in-vivo extracellular pH in rat brain gliomas [3], Electron spin resonance (ESR), which is operated at a lower resonance, has been adapted for in-vivo pH measurements because it provides a sufficient RF penetration for deep body organs [28], The non-destructive determination of tissue pH using near-infrared diffuse reflectance spectroscopy (NIRS) has been employed for pH measurements in the muscle during... 

Metal complexes like lanthanide chelates (mainly europium or terbium), ruthenium phenanthrolines or bipyridyls, and platinum porphyrins can be used as fluorescent labels for biomolecules. Their long decay times are perfectly suited for a detection by time-resolved imaging, and the labeled target molecules can be used for the determination of intracellular recognition processes or for the screening of DNA and protein arrays. Ratiometric lifetime-based imaging methods in combination with sophisticated data acquisition and evaluation tools can substantially contribute to the development... 

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