High refractive index substrates for fluorescence microscopy of biological interfaces with high
            <i>z</i>
            contrast

Ajo‐Franklin, Caroline M.; Kam, Lance C.; Boxer, Steven G.

doi:10.1073/pnas.241208698

Cited by 29 publications

(22 citation statements)

References 35 publications

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“…The absorption transition dipole orientation was allowed to vary initially before being fixed at 60° relative to bilayer normal. This value is quite different from the expected value near 90° (Ajo-Franklin, Kam et al 2001). This difference may be because the DNA-tethered lipid membranes are more prone to fluctuations than a bilayer directly on a solid support, although deviations from expected values have been reported for other dyes in supported bilayers (Sund, Swanson et al 1999).…”

Section: Methodscontrasting

confidence: 82%

DNA-tethered membranes formed by giant vesicle rupture

Chung

Lowe

Chan

et al. 2009

Journal of Structural Biology

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105

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We have developed a strategy for preparing tethered lipid bilayer membrane patches on solid surfaces by DNA hybridization. In this way, the tethered membrane patch is held at a controllable distance from the surface by varying the length of the DNA used. Two basic strategies are described. In the first, single-stranded DNA strands are immobilized by click chemistry to a silica surface, whose remaining surface is passivated to prevent direct assembly of a solid supported bilayer. Then giant unilamellar vesicles (GUVs) displaying the antisense strand, using a DNA-lipid conjugate developed in earlier work (Chan, Lengerich et al. 2008), are allowed to tether, spread and rupture to form tethered bilayer patches. In the second, a supported lipid bilayer displaying DNA using the DNA-lipid conjugate is first assembled on the surface. Then GUVs displaying the antisense strand are allowed to tether, spread and rupture to form tethered bilayer patches. The essential difference between these methods is that the tethering hybrid DNA is immobile in the first, while it is mobile in the second. Both strategies are successful; however, with mobile DNA hybrids as tethers, the patches are unstable, while in the first strategy stable patches can be formed. In the case of mobile tethers, if different length DNA hybrids are present, lateral segregation by length occurs and can be visualized by fluorescence interference contrast microscopy making this an interesting model for interactions that occur in cell junctions. In both cases, lipid mobility is high and there is a negligible immobile fraction. Thus, these architectures offer a flexible platform for the assembly of lipid bilayers at a well-defined distance from a solid support. KeywordsTethered lipid membrane; surface-immobilized DNA; Giant unilamellar vesicles Supported lipid bilayers (SLBs) have been widely used as a model for cell membranes (Sackmann 1996;Chan and Boxer 2007) and to investigate membrane components including proteins in a simpler context apart from the complex cellular environment. SLBs are assembled by Langmuir-Blodgett techniques or spontaneous fusion of unilamellar vesicles on carefully prepared surfaces, usually hydrophilic solid supports, such as glass (Seu, Pandey et al. 2007), silica, mica (Richter, Berat et al. 2006), or TiO 2 (Rossetti, Bally et al. 2005). Although SLBs have the advantages of simple formation, easy handling and are well-suited for investigation by a suite of surface sensitive methods due to their planar geometry, the close proximity of the lower leaflet to the solid support often leads to unfavorable interactions with integral membrane proteins. Recognizing this limitation, many groups have described methods to separate the © 2009 Elsevier Inc. All rights reserved. *Corresponding Author: Tel.: 1-650-723-4482; Fax: 1-650-723-4817; E-mail: sboxer@stanford.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early versi...

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Section: Methodscontrasting

confidence: 82%

DNA-tethered membranes formed by giant vesicle rupture

Chung

Lowe

Chan

et al. 2009

Journal of Structural Biology

Self Cite

105

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“…The best substrates are fused silica [12,61], borosilicate glass [12,62], mica [63,64], and oxidized silicon [12]. Attempts have been made to deposit supported bilayers on single crystals of TiO 2 and SrTiO 2 as well as on thin films of SiO 2 on LiNbO 3 crystals [65][66][67]. Thin films can be used as solid supports as observed with TiO 2 [68][69][70], indium-tin-oxide [71,72], gold [73,74], silver [75], and platinum [76].…”

Section: Solid Supported Lipid Bilayersmentioning

confidence: 99%

Solid supported lipid bilayers: From biophysical studies to sensor design

Castellana

Cremer

2006

Surface Science Reports

1,003

1,009

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The lipid bilayer is one of the most eloquent and important self-assembled structures in nature. It not only provides a protective container for cells and sub-cellular compartments, but also hosts much of the machinery for cellular communication and transport across the cell membrane. Solid supported lipid bilayers provide an excellent model system for studying the surface chemistry of the cell. Moreover, they are accessible to a wide variety of surface-specific analytical techniques. This makes it possible to investigate processes such as cell signaling, ligand-receptor interactions, enzymatic reactions occurring at the cell surface, as well as pathogen attack. In this review, the following membrane systems are discussed: black lipid membranes, solid supported lipid bilayers, hybrid lipid bilayers, and polymer cushioned lipid bilayers. Examples of how supported lipid membrane technology is interfaced with array based systems by photolithographic patterning, spatial addressing, microcontact printing, and microfluidic patterning are explored. Also, the use of supported lipid bilayers in microfluidic devices for the development of lab-on-a-chip based platforms is examined. Finally, the utility of lipid bilayers in nanotechnology and future directions in this area are discussed.

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“…This technique exploits the evanescent field that is produced at an interface of two media with different refractive indexes. It can achieve resolution of 30–100 nm [99]. An inherit issue of this technique, is that it can only study emission at (or close to) the interface [100], therefore it cannot be used for imaging inside the cell.…”

Section: Methods: Individual and Complementary Biophysical Measuresmentioning

confidence: 99%

Single cell spectroscopy: Noninvasive measures of small-scale structure and function

Mousoulis

Reiter

et al. 2013

Methods

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The advancement of spectroscopy methods attained through increases in sensitivity, and often with the coupling of complementary techniques, has enabled real-time structure and function measurements of single cells. The purpose of this review is to illustrate, in light of advances, the strengths and the weaknesses of these methods. Included also is an assessment of the impact of the experimental setup and conditions of each method on cellular function and integrity. A particular emphasis is placed on noninvasive and nondestructive techniques for achieving single cell detection, including nuclear magnetic resonance, in addition to physical, optical, and vibrational methods.

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High refractive index substrates for fluorescence microscopy of biological interfaces with high z contrast

Cited by 29 publications

References 35 publications

DNA-tethered membranes formed by giant vesicle rupture

DNA-tethered membranes formed by giant vesicle rupture

Solid supported lipid bilayers: From biophysical studies to sensor design

Single cell spectroscopy: Noninvasive measures of small-scale structure and function

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