Single-Bead Fluorescence Microspectroscopy:  Detection of Self-Quenching in Fluorescence-Labeled Resin Beads

Yan, Bing; Martin, and Paul C.; Lee, Jumi

doi:10.1021/cc9800069

Cited by 29 publications

(33 citation statements)

References 26 publications

(22 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consistent with previous observations [12] [14], we believe that the lack of fluorescence in the center of bead 26 is due to self-quenching by proximal chromophores, when all sites are occupied. Active sites are closer together in the center of the bead 8 ).…”

supporting

confidence: 92%

“…Also, it is less susceptible to self-quenching, when several fluorophores are in close proximity because of its large Stokes× shift (ca. 200 nm) [12]. Preparation of Fluorescent Dendrimers.…”

mentioning

confidence: 99%

“…The phenomenon of fluorescence quenching in epifluorescence microscopy has been addressed in both whole beads [12] and slices made with a microtome [14], as well as in confocal microscopy [14]. We did not anticipate that this phenomenon would be important, when the dye was encapsulated in a dendrimer, but in the absence of the dendrimer, we wanted to avoid the issue.…”

mentioning

confidence: 99%

“…We also prepared beads with coumarin 5 and cyclohexanecarboxylic acid in 1 : 9 and 1 : 27 proportions, 28 and 29, respectively. The role of the cyclohexanecarboxylic acid, a non-fluorescent molecule, is to dilute the fluorophore throughout the resin to prevent self-quenching by proximal fluorophores on solid support [12] [14] [15]. Note that all the beads were derivatized under very similar conditions.…”

mentioning

confidence: 99%

See 3 more Smart Citations

Spatially Resolved Derivatization of Solid-Phase-Synthesis Beads with Fluorescent Dendrimers: Creation of Localized Microdomains

Cardona

Jannach

Huang

et al. 2002

HCA

View full text Add to dashboard Cite

supporting

confidence: 92%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Spatially Resolved Derivatization of Solid-Phase-Synthesis Beads with Fluorescent Dendrimers: Creation of Localized Microdomains

Cardona

Jannach

Huang

et al. 2002

HCA

View full text Add to dashboard Cite

“…Considering the fact that the fluorescence of a dye with a large Stokes shift was not selfquenched on the resin bead, 12 the quenching effect of resin A may be due to physical confinement of the dyes on to a small resin rather than due to a photochemical quenching. This phenomenon opened up the chance to study an in situ kinetic behavior of this resin without any sampling process, which facilitated data collection in a short time.…”

mentioning

confidence: 99%

Single Resin Bead Kinetics Using Real Time Fluorescence Meaurements

Parikh

Kim²,

Chang

2002

Bulletin of the Korean Chemical Society

View full text Add to dashboard Cite

Fluorescence techniques have been used widely in chemistry and biology for over a century.1 With the advent of combinatorial solid phase synthesis, a variety of fluorescence techniques for resin beads have been developed and adopted for broad applications, which include encoding/decoding, 2 fluorescent molecule binding assays, 3 and catalytic activity detection. 4 The split-and-pool method, another important technique in combinatorial chemistry, facilitates the synthesis of millions of molecules in a one-bead-one-compound library format; in principle, each single bead of resin acts as a separate microreactor. Although conventional screenings of these split-and-pool libraries have usually been performed as mixtures, a segregation approach of each single resin has been demonstrated recently, avoiding complicated data analysis. 5 To simplify the handling of a single resin, the researchers necessarily utilized a super size bead (a 500 µm diameter Rapp PS (polystyrene) bead, about 5-fold larger in diameter than an ordinary synthetic resin) with the aid of bead arraying tools.The problem with the kinetics of solid phase reactions is that it demands, in many cases, if not all, tedious sample preparation steps including filtering, washing and transferring. While single resin bead FT-IR spectra have been utilized successfully to quantify solid phase reaction rates, 6 no comparable fluorescence kinetics has yet been demonstrated. Herein we report the first example of single resin bead kinetics using continuous measurement of fluorescence generation on an activated ester resin.Activated esters on solid support have been developed as convenient labeling reagents, especially for amine nucleophiles.7-9 Most of the reported functionalities, such as nitrophenol, N-hydroxysuccinimide, HOBt (1-hydroxybenzotriazole) and Kaiser oxime, have been attached to a polystyrene solid support by a Friedel-Craft reaction 10 or to a thiol resin by a maleimide linker, 9 limiting the selection of resin compositions. To overcome this limitation, we utilized the well-established amide bond formation to couple tetrafluoro-4-hydroxy benzoic acid with an aminomethyl resin (1), which has a broad material selection, creating a novel tetrafluorophenol resin (2).11 For kinetic studies under various conditions, an environment-insensitive high quantum yield fluorescence molecule, 4-acetamino-1,8-naphthalimide (3), 7 was chosen to prepare the activated ester resin (A). 11When reacted with a small amount of benzylamine (B) in various solvents, resin A gave a pure product (C) without work-up or purification, and the amount of generated product could be quantified by fluorescence measurement. Similarly obtained fluorescence data was previously used to calculate the second order reaction rate constant of nitrophenol resin, after removal of the resin by filtration followed by a sample dilution. 7One important observation was that the fluorescence was quenched almost completely in the resin-bound molecule, while the released product C showed a bright fluoresce...

show abstract

Site Distribution in Resin Beads as Determined by Confocal Raman Spectroscopy

Kress

Rose

Frey

et al. 2001

Chemistry A European J

View full text Add to dashboard Cite

Scanning confocal Raman spectroscopy was used to study the distribution of reactive sites within a resin bead used for solid-phase synthesis. The distribution of NH2 groups in aminomethylated polystyrene resin (APS) was determined by doping with varying amounts of 4-cyanobenzoic acid. The extent of loading was determined by both elemental analysis and ninhydrin assays. The spatial distribution of the coupled 4-cyanobenzamide within the bead was determined to an in-plane resolution of 1 microm and depth resolution of about 4 microm, using the strong Raman CN stretching vibrational transition at 2230 cm(-1). Dry and swollen beads were studied and the distribution was found to be essentially uniform throughout the bead in all cases.

show abstract

Single-Bead Fluorescence Microspectroscopy: Detection of Self-Quenching in Fluorescence-Labeled Resin Beads

Cited by 29 publications

References 26 publications

Spatially Resolved Derivatization of Solid-Phase-Synthesis Beads with Fluorescent Dendrimers: Creation of Localized Microdomains

Spatially Resolved Derivatization of Solid-Phase-Synthesis Beads with Fluorescent Dendrimers: Creation of Localized Microdomains

Single Resin Bead Kinetics Using Real Time Fluorescence Meaurements

Site Distribution in Resin Beads as Determined by Confocal Raman Spectroscopy

Contact Info

Product

Resources

About