Biosensors जटिल, जैविक वातावरण के साथ इंटरफेस और अत्यधिक विशिष्ट सतह के संशोधन के माध्यम से सेंसर से जुड़ी जांच के साथ अत्यधिक संवेदनशील सेंसर के संयोजन द्वारा लक्षित पता लगाने के प्रदर्शन. यहाँ, हम बायोटिन साथ सिलिका ऑप्टिकल सेंसर silane युग्मन एजेंट का उपयोग करने के लिए सेंसर और जैविक पर्यावरण को पाटने की सतह functionalization प्रदर्शित करता है.
In order to interface with biological environments, biosensor platforms, such as the popular Biacore system (based on the Surface Plasmon Resonance (SPR) technique), make use of various surface modification techniques, that can, for example, prevent surface fouling, tune the hydrophobicity / hydrophilicity of the surface, adapt to a variety of electronic environments, and most frequently, induce specificity towards a target of interest.1-5 These techniques extend the functionality of otherwise highly sensitive biosensors to real-world applications in complex environments, such as blood, urine, and wastewater analysis.2,6-7 While commercial biosensing platforms, such as Biacore, have well-understood, standard techniques for performing such surface modifications, these techniques have not been translated in a standardized fashion to other label-free biosensing platforms, such as Whispering Gallery Mode (WGM) optical resonators.8-9
WGM optical resonators represent a promising technology for performing label-free detection of a wide variety of species at ultra-low concentrations.6,10-12 The high sensitivity of these platforms is a result of their unique geometric optics: WGM optical resonators confine circulating light at specific, integral resonance frequencies.13 Like the SPR platforms, the optical field is not totally confined to the sensor device, but evanesces; this “evanescent tail” can then interact with species in the surrounding environment. This interaction causes the effective refractive index of the optical field to change, resulting in a slight, but detectable, shift in the resonance frequency of the device. Because the optical field circulates, it can interact many times with the environment, resulting in an inherent amplification of the signal, and very high sensitivities to minor changes in the environment.2,14-15
To perform targeted detection in complex environments, these platforms must be paired with a probe molecule (usually one half of a binding pair, e.g. antibodies / antigens) through surface modification.2 Although WGM optical resonators can be fabricated in several geometries from a variety of material systems, the silica microsphere is the most common. These microspheres are generally fabricated on the end of an optical fiber, which provides a “stem” by which the microspheres can be handled during functionalization and detection experiments. Silica surface chemistries may be applied to attach probe molecules to their surfaces; however, traditional techniques generated for planar substrates are often not adequate for these three-dimensional structures, as any changes to the surface of the microspheres (dust, contamination, surface defects, and uneven coatings) can have severe, negative consequences on their detection capabilities. Here, we demonstrate a facile approach for the surface functionalization of silica microsphere WGM optical resonators using silane coupling agents to bridge the inorganic surface and the biological environment, by attaching biotin to the silica surface.8,16 Although we use silica microsphere WGM resonators as the sensor system in this report, the protocols are general and can be used to functionalize the surface of any silica device with biotin.
प्रोटोकॉल में वर्णित है, हम एक आवास मंच बनाया है जिसके द्वारा functionalization प्रक्रिया के दौरान उनके उपजी सिलिका microspheres के परिवहन के लिए. इस आवास मंच सतह संदूषण और क्षति कि microsphere विभिन्न functionalization प्रक्रिया के दौरान ?…
The authors have nothing to disclose.
लेखकों को आभार इस प्रोटोकॉल विकसित किया गया था समय के दौरान समर्थन के लिए दक्षिणी कैलिफोर्निया विश्वविद्यालय में प्रो आंद्रे अरमानी स्वीकार करते हैं. इस काम के प्रारंभिक विकास के लिए अनुदान राष्ट्रीय विज्ञान फाउंडेशन द्वारा प्रदान किया गया [085,281 और 1,028,440] और एनआईएच के निदेशक का नया अन्वेषक पुरस्कार कार्यक्रम के माध्यम से स्वास्थ्य के राष्ट्रीय संस्थान [1DP2OD007391-01. अतिरिक्त जानकारी पर उपलब्ध है http://web.missouri.edu/ ~ / hunthk .
Name of the reagent | Company | Catalogue number | Comments |
Methanol | Fisher | 67-56-1 | ACS grade |
Sulfuric Acid | Fisher | 8014-95-7 | Fuming |
Hydrogen Peroxide | Fisher | 7722-84-1 | 30 wt % |
Aminopropyltrimethoxysilane | Fisher | 13822-56-5 | |
NHS-biotin EZ linker | Pierce | 20217 | |
Dimethylsulfoxide | Fisher | 67-68-5 | Anhydrous |
Fluorescein Isothiocyanate | Pierce | 46425 | |
Phosphate Buffered Saline | Fisher | 7647-14-5 | Powder concentrate |
Sodium Bicarbonate Buffer | Fisher | NC0099321 | |
Texas Red – Avidin Conjugate | Pierce | A820 | |
Optical Fiber | Newport | F-SC | |
Fiber Stripper | Fiber Instrument Sales | NN-175 | No-Nik 175 um stripper |
Kimwipes | Fisher | 06666A | |
Bare Fiber Cleaver | Ilsintech | Cl-03A | |
Glass Microscope Slides | Fisher | 12-550B | |
Polypropylene Vials | Fisher | 03-341-75A | 60 mL, hinged cap |
Incubating Rocker | VWR | 12620-910 | |
Vacuum Desiccator | Fisher | 08-594-15B |