This work was supported by a Grant-in-Aid for Scientific Research No. 22750122 (S.H.), No. 26288099 (T.Y.), and No. 23350050 (Y.T.) of the Japan Society for the Promotion of Science. S.H. is grateful for The Kurata Memorial Hitachi Science and Technology Foundation. The research reported in this publication was supported by the King Abdullah University of Science and Technology (S.H.).

1. Síntese de monofuncional e poli Bifuncional (THF) <ol><li> Poli monofuncional (THF) <ol><li> Chama seco num balão de fundo redondo de 2 tubuladuras de 100 ml. Vácuo e encher o balão com azoto (3 ciclos). </li><li> Adicionar tetra-hidrofurano destilado (THF) (50 ml) ao balão. Colocar o balão num banho de água a 20 ° C e a temperatura equilibrar. </li><li> Adicionar triflato de metilo (0,5 mmol) ao frasco por uma seringa. Agita-se a mistura durante 5-10 min a 20 ° C. </li><li> Adiciona-se N -fenil pirrolidina (4-6 equiv.) Ao frasco por uma seringa. Agita-se a mistura durante 30-60 minutos. </li><li> Remover completamente o solvente sob pressão reduzida (cerca de 100 Torr). Dissolve-se o resíduo em 3-5 ml de acetona. Adicionar a solução de acetona em 300-500 ml de n-hexano. Filtra-se o precipitado e secá-lo sob pressão reduzida. </li></ol></li><li> Poli bifuncional (THF) <ol><li> Chama seco num balão de fundo redondo de 2 tubuladuras de 100 ml. Vaente e encher o balão com azoto (3 ciclos). </li><li> Adicionar THF destilado (50 ml) ao balão. Colocar o balão num banho de água a 20 ° C e a temperatura equilibrar. </li><li> Adicionar anidrido triflico (0,3 mmol) ao frasco por uma seringa. Agita-se a mistura durante 5-10 min a 20 ° C. </li><li> Adiciona-se N -fenil pirrolidina (4-6 equiv.) Ao frasco por uma seringa. Agita-se a mistura durante 30-60 minutos. </li><li> Remover completamente o solvente sob pressão reduzida (cerca de 100 Torr). Dissolve-se o resíduo em 3-5 ml de acetona. Adicionar a solução de acetona a 300-500 ml de n-hexano. Filtra-se o precipitado e secá-lo sob pressão reduzida. </li></ol></li></ol> em forma de 8 2. Síntese de Perileno diimida-incorporada por estrelas 4-armada e dicíclico Poly (THF) <ol><li> Armado poli estrela (THF) <ol><li> troca iônica <ol><li> Dissolve-se o sal de di-imida perileno tetracarboxilato de sódio em água (10 mg / ml, 150 ml). Dissolverpoli monofuncional (THF) em acetona (160 mg / ml, 4 ml). Adicionar gota a gota, a solução de acetona na solução aquosa vigorosamente agitada. Recolhe-se o precipitado formado por filtração. </li><li> Repetir o procedimento acima com o precipitado recuperado (2.1.1.1) quatro vezes. </li></ol></li><li> fixação covalente <ol><li> Dissolve-se o precipitado obtido em tolueno (5 mg / ml). Refluxo a solução durante 4 horas. </li><li> Remover completamente o solvente sob pressão reduzida (cerca de 100 Torr). Filtra-se o resíduo através de um tampão de gel de sílica com n-hexano / acetona (2/1 vol / vol). Adicionar a solução em água arrefecida com gelo (300-500 ml) para precipitar o produto. Recolher o precipitado por filtração. </li></ol></li></ol></li><li> Poli em forma de oito dicíclico (THF) <ol><li> troca iônica <ol><li> Dissolve-se o sal de di-imida perileno tetracarboxilato de sódio em água (6 mg / ml, 50 ml). Dissolve-se poli bifuncional (THF) (0,5 g) Em 30-50 ml de acetona. Adicionar gota a gota, a solução de acetona na solução aquosa vigorosamente agitada a 0 ° C. Recolhe-se o precipitado formado por filtração. </li><li> Repetir o procedimento acima com o precipitado recuperado (2.2.1.1). </li></ol></li><li> fixação covalente <ol><li> Dissolve-se o precipitado obtido em tolueno (0,05 g / L). Refluxo a solução durante 4 horas. </li><li> Remover completamente o solvente sob pressão reduzida (cerca de 100 Torr). Adicionar tolueno para dissolver parcialmente o resíduo. Re-precipitar em 300-500 ml de n-hexano. </li><li> Filtra-se o precipitado formado por meio de um tampão de gel de sílica com n-hexano / acetona (2/1 vol / vol). Re-precipitar em 300-500 ml de água. </li><li> Purifica-se o precipitado que se formou por cromatografia em coluna utilizando 18 um gel de poliestireno. Purificar o produto bruto por cromatografia de permeação em gel preparativa (GPC) 19 com um eluente de CHCl3 a REMsubprodutos ove, monitorando índice de refração (RI) e detectores de UV. </li></ol></li></ol></li></ol> 3. única molécula de fluorescência de imagem Experiment <ol><li> Preparação de amostra <ol><li> Limpeza de lamelas de microscópio <ol><li> Coloque a tampa mm microscópio No. 1.5 24 x 24 desliza em uma tina de coloração. </li><li> Adicionar solução de hidreto de potássio 1 M (100 ml) no frasco e sonicado durante 15 minutos. Decantar a solução de hidróxido de potássio por decantação e lavar as lamelas com água ultra-pura por várias vezes. Adicionar etanol de qualidade espectroscópica (100 ml) no frasco e sonicado durante 15 minutos. </li><li> Deitar fora o etanol por decantação e lavar as lamelas com água ultra-pura por várias vezes. Após o vazamento para fora da água ultra-pura por decantação, repita o passo 3.1.1.2. </li><li> Adicione a água ultra-pura para o frasco e sonicate para 15 min. Lavar as lamelas com água ultra-pura por diversas vezes. Retire as lamelas do frasco por uma pinça de plástico e secá-las por qualquer ar seco ou azoto seco. </li></ol></li><li> Preparação de amostras de massa fundida de polímero 14,15 <ol><li> Adicionar 100 ul de poli linear não marcado (THF) num frasco de vidro e aquecê-la a uma temperatura acima do ponto de fusão (cerca de 25 ° C) usando um secador de cabelo. </li><li> Dissolve-se o polímero incorporado-fluoróforo (linear, em estrela de 4 ramos, cíclico, ou dicíclico em forma de oito sintetizado em 2,1 e 2,2) em clorofórmio (1 ml, 10 -6 M). Adicionar 1 ml de solução aos 100 ul da massa fundida do poli linear não marcado (THF). </li><li> Depois de se misturar cuidadosamente a amostra com uma ponta de pipeta, evapora-se o clorofórmio por aquecimento da amostra, utilizando um secador. NOTA: Este fornece uma massa fundida do poli linear não marcado (THF) contendo 10 -8 M de os polímeros fluoróforos incorporados. </li><li> Tomar 10 ul da amostra, utilizando uma micro-pipeta e dROP-lo em uma lamela limpa. Dito de outra lamela limpo na amostra e sanduíche da amostra entre as duas lamelas. </li><li> Pressione suavemente a amostra usando uma pinça de plástico. </li></ol></li></ol></li><li> Wide-campo de configuração imagens de fluorescência 15 <ol><li> Introdução de um laser de excitação (488 nm) para a porta de trás do microscópio invertido <ol><li> Insira um filtro de excitação de banda e polarizador para o caminho do feixe. </li><li> Expandir o feixe de cerca de 1 cm de diâmetro por um expansor de feixe. </li><li> Insira uma placa de quarto de onda para o caminho do feixe. Definir o eixo óptico da placa de onda a 45 graus em relação ao do polarizador. Alternativamente, insira um compensador de Berek e definir o atraso óptico para? / 4. </li><li> Inserir um diafragma no caminho do feixe de excitação para ajustar o tamanho do feixe. </li><li> Antes de introduzir o feixe de laser na porta traseira do microsco óptico invertidoPE, inserir uma lente de focagem (lentes plano-convexo, o comprimento focal ≈ 300 mm) a uma posição em que o feixe de laser para fora da lente objectiva é colimada. </li></ol></li><li> Depois de reflectir o feixe de laser utilizando um espelho dicróico montado sobre um cubo de filtro, introduzir o feixe de laser para a amostra através de uma lente objectiva de alta abertura numérica (NA) (por exemplo, NA 1.3, uma ampliação de 100X, de imersão em óleo). </li><li> Anexar um aquecedor objectivo para a lente objectiva e ajustar a temperatura a 30 ° C. </li><li> Montagem da amostra sobre a fase de microscópio invertido <ol><li> Deixe cair uma gota de óleo de imersão na lente objetiva e montar a amostra sobre o estado microscópio. </li><li> Assegure-se que a espessura da amostra de cerca de 10 um é obtido através da verificação da posição axial da superfície de fundo e de topo da amostra. </li><li> Ajustar a focagem do microscópio, para alguns micrómetros acima da superfície inferior da amostra. </li></ol></li&gt; <li> Obter luz de excitação polarizada circularmente sob a lente objetiva <ol><li> Insira um polarizador para o raio laser colimado para fora da lente objetiva. </li><li> Grave a intensidade do laser transmitida através do polarizador através da inserção de um medidor de energia após o polarizador. Grave a potência do laser transmitida em diferentes ângulos de polarização através da rotação do polarizador. </li><li> Se a potência do laser transmitida não é constante em todos os ângulos de polarização, gire ligeiramente a placa de quarto de onda ou Berek compensador inserido no caminho do feixe de excitação. </li><li> Repita o passo 3.2.5.2 e 3.2.5.3 até que a constante potência do laser transmitido é obtido em todos os ângulos de polarização. Certifique-se de que a luz polarizada circularmente é obtido na amostra. </li></ol></li><li> Configuração do EM (multiplicador de elétrons) -Charge coupled device (CCD) <ol><li> Fixe a câmara EM-CCD à porta lateral do microscópio e conectá-lo para tele software de aquisição de imagem. </li><li> Se necessário, sincronizar a exposição da câmara para um obturador mecânico ou filtro sintonizável acústico-óptico inserido no caminho do feixe de excitação através do envio do Transistor-Transistor Logic (TTL) sinais gerada pela câmara EM-CCD para os dispositivos. Alternativamente, sincronizar a exposição da câmara para a saída do laser, enviando os sinais TTL gerados pela câmara CCD EM-ao laser. NOTA: A última opção é aplicável somente quando um laser de estado sólido, cuja potência de saída pode ser modulada pela entrada lógica transístor-transístor (TTL), sinais é utilizado para a experiência. </li><li> Aplicar um ganho EM (tipicamente, aproximadamente, 300) para a câmara CCD usando software que controla a câmara, a fim de obter uma imagem de fluorescência de alta qualidade do único fluoróforo. </li><li> Definir uma região de interesse (ROI) (tipicamente 128 x 128 pixels no centro do campo de vista), usando software que controla a câmara. NOTA: Isto permite a imagiexperimentos ng em taxas de quadro de 100-200 Hz no modo de transferência de quadro, o que é necessário para visualizar o movimento das cadeias de polímero incorporado-fluoróforo na amostra de fusão. </li></ol></li></ol></li><li> A execução do experimento <ol><li> Otimizar as condições experimentais <ol><li> Ajustar a área de iluminação da amostra a cerca de 20 um de diâmetro usando o diafragma inserido no caminho do feixe de excitação. </li><li> Definir a potência do laser de excitação na amostra a 4-8 mW manualmente selecionando um filtro apropriado de densidade neutra (ND) inserido no caminho do feixe de excitação. NOTA: Este fornece a potência do laser médio de 1 - 2 cm -2 kW na amostra. </li><li> Gravação de imagens de fluorescência da amostra nas taxas de quadro de 100-200 Hz. Se a intensidade de fluorescência obtidos a partir dos polímeros individuais incorporadas-fluoróforo é demasiado baixa, aumentar gradualmente a potência de excitação utilizando the ND filtro até atingir cerca de 100 mW na amostra. </li><li> Se a qualidade da imagem de fluorescência única molécula ainda não é satisfatória, verificar as impurezas de fluorescência na amostra através da gravação de imagens de fluorescência de uma massa fundida pura do poli não marcado (THF). No caso de um fundo de fluorescência elevada é observada, usar poli diferente não marcado (THF). </li><li> Se a densidade do local de fluorescência obtidos a partir dos polímeros incorporada fluororo na massa fundida é demasiado elevado para isolá-los (o que provoca erros na análise do movimento difusivo) espacialmente, diminuir a concentração dos polímeros incorporada fluororo na amostra até espacialmente são observados pontos isolados. </li><li> Se a densidade do local de fluorescência obtidos a partir dos polímeros incorporada fluororo no estado fundido é muito baixa (isto causa um baixo rendimento do experimento de imagem), aumentar a concentração dos polímeros incorporada fluororo na amostra até uma aproprdensidade iate do local de fluorescência é atingido. </li><li> Se as imagens de fluorescência obtidos a partir dos polímeros incorporou-fluoróforo no estado fundido são borradas, aumentar as taxas de quadro de aquisição de imagem. NOTA: Isso muitas vezes requer um ROI menor, normalmente 64 x 64 pixels. </li></ol></li></ol></li><li> Aquisição de imagem <ol><li> Uma vez que as condições experimentais são optimizadas, deixar a amostra montada na platina do microscópio para uma hora de modo a que a amostra atinge condições de equilíbrio. </li><li> Grave 500 - 1000 sequências de imagens de fluorescência dos polímeros incorporada fluororo no estado de fusão a 100 - 200 Hz taxa de quadros. Se o formato de arquivo padrão não é TIFF, converter todas as sequências de imagens para o formato TIFF. </li></ol></li></ol> 4. Análise do movimento difusivo <ol><li> Deslocamento quadrático médio de análise (MSD) <ol><li> Cortar as sequências de imagens de fluorescência de tal formaque cada sequência de imagens contém um polímero usando difusão de imagem software de processamento incorporadas-fluoróforo única e bem focada, como ImageJ. </li><li> Quando as sequências de imagens recortadas conter mais de 10 quadros, dividir as sequências de imagens em várias sequências de tal forma que cada sequência é composta por 10 quadros. </li><li> Determinar as posições das moléculas em cada sequências de imagem com precisão por encaixe Gaussiana bidimensional das imagens. </li><li> Determinar o coeficiente de difusão (D) das moléculas individuais através da análise quadrático médio deslocamento (MSD) das trajectórias de difusão (isto é, posições dependentes do tempo da molécula) utilizando uma equação 20 <img alt="equação1" src="/files/ftp_upload/54503/54503equation1.jpg" /> onde x i e y i são as posições da molécula no quadro de imagem i, e n indica o número de quadro com o lapso de tempo At do quadro I. </li><li> Traçar os coeficientes de difusão emum histograma de frequência. NOTA: Tipicamente, o histograma é construído a partir de mais do que 100 moléculas. </li></ol></li><li> Função de distribuição cumulativa de análise (CDF) NOTA: Um CDF, P (R 2, i Δ t) corresponde à probabilidade cumulativa de encontrar as moléculas de difusão dentro de um raio R a partir da origem, após um certo lapso de tempo i ô t. <ol><li> Calcule o deslocamento quadrado que ocorre durante defasagens de 1Δt, 2Δt, ····, iΔt para todas as trajetórias de difusão obtidos em 4.1.3. NOTA: Estes cálculos dar m total de i-quadrado deslocamentos para atrasos no momento da iΔt. </li><li> Calcule números dos deslocamentos-quadrado (l i) no prazo total de conjunto de dados m i que são menores que r 2 em diferentes valores de r 2 (0 &lt;r2 &lt;∞). Normalizada l i vs r 2<html> parcelas correspondem a CDF, P (R 2, iΔt). </li></ol></li><li> Análise de CDFs com modelos de difusão distintas Nota: Os CDFs obtidos são ajustados por modelos de difusão distintas; modelo homogéneo difusão, vários modos de difusão na qual a distribuição D é descrita por uma distribuição Gaussiana (modelo gaussiano único), e vários modos de difusão na qual a distribuição D é descrita por múltiplos Gaussiana (múltiplos modelo gaussiano). <ol><li> No modelo de difusão homogénea, determinam uma média de D, ajustando a CDF usando uma equação 21 <img alt="Equation2" src="/files/ftp_upload/54503/54503equation2.jpg" /> NOTA: Qualquer desvio a partir da equação sugere a difusão heterogênea da molécula. </li><li> No modelo gaussiano único, determinar a distribuição de probabilidade de D descrito por uma Gaussiana (f (D)), através da montagem do CDF utilizando 15 on3 &quot;src =&quot; / files / ftp_upload / 54503 / 54503equation3.jpg &quot;/&gt; <img alt="Equation4" src="/files/ftp_upload/54503/54503equation4.jpg" /> em que A, W, e D são 0 a amplitude, a largura, e o centro de Gauss. </li><li> No modelo gaussiano dupla, determinar a distribuição de probabilidade de ordem j de componente D descrito por uma Gaussiana (f (D)), através da montagem do CDF usando 14 <img alt="Equation5" src="/files/ftp_upload/54503/54503equation5.jpg" /> <img alt="Equation6" src="/files/ftp_upload/54503/54503equation6.jpg" /> Um onde j é a fracção de cada componente de difusão, e α J, W J, D e J são 0 a amplitude, a largura, e o centro de ordem j componente da Gaussiana. </li></ol></li><li> Cálculo do distri probabilidade teóricabuição de coeficiente de difusão NOTA: As distribuições de probabilidade de D que ocorrem devido a erros estatísticos (p (D) d D) são calculados para os diferentes modelos de difusão; modelo homogéneo difusão, vários modos de difusão na qual a distribuição D é descrita por uma distribuição Gaussiana (modelo gaussiano único), e vários modos de difusão na qual a distribuição D é descrita por múltiplos Gaussiana (múltiplos modelo gaussiano). <ol><li> No modelo de difusão homogénea, calcular a distribuição de probabilidade estatística de D utilizando uma equação 22 <img alt="Equation7" src="/files/ftp_upload/54503/54503equation7.jpg" /> onde N é o número dos pontos de dados em uma trajectória de difusão (N = 10, ver 4.1.2), D 0 é o coeficiente de difusão médio (determinado por análise do CDF, ver 4.2.3.1), e D é a obtida experimentalmente coeficiente de difusão para uma trajetória individual. </li><li> Noúnico modelo de difusão Gaussian, calcular a distribuição de probabilidade estatística de D usando uma equação 15 <img alt="Equation8" src="/files/ftp_upload/54503/54503equation8.jpg" /> onde f (D) indica a distribuição de probabilidade de D determinado por análise CDF (ver 4.2.3.2), e D 0 é o coeficiente de difusão médio (determinado por análise do CDF, ver 4.2.3.2). </li><li> No modelo de difusão Gaussian casal, calcular a distribuição de probabilidade estatística de D usando uma equação 14 <img alt="Equation9" src="/files/ftp_upload/54503/54503equation9.jpg" /> onde f (D J) indica a distribuição de probabilidade de o componente de ordem j D (D J) determinada pela análise de CDF (ver 4.2.3.3), e D 0j é o coeficiente de difusão médio de j th componente (determinado pelo CDF análise, ver 4.2.3.3). </li></ol></li></ol>

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Soc. 122 (40), 9592-9599 (2000).</li><li>Yamamoto, T., Tezuka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topological+polymer+chemistry:+a+cyclic+approach+toward+novel+polymer+properties+and+functions.">Topological polymer chemistry: a cyclic approach toward novel polymer properties and functions.</a> Polym. Chem. 2 (9), 1930-1941 (2011).</li><li>Habuchi, S., Fujiwara, S., Yamamoto, T., Tezuka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-molecule+imaging+reveals+topological+isomer-dependent+diffusion+by+4-armed+star+and+dicyclic+8-shaped+polymers.">Single-molecule imaging reveals topological isomer-dependent diffusion by 4-armed star and dicyclic 8-shaped polymers.</a> Polym. Chem. 6 (22), 4109-4115 (2015).</li><li>Habuchi, S., Fujiwara, S., Yamamoto, T., Vacha, M., Tezuka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-Molecule+Study+on+Polymer+Diffusion+in+a+Melt+State:+Effect+of+Chain+Topology.">Single-Molecule Study on Polymer Diffusion in a Melt State: Effect of Chain Topology.</a> Anal. Chem. 85 (15), 7369-7376 (2013).</li><li>Habuchi, S., Satoh, N., Yamamoto, T., Tezuka, Y., Vacha, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multimode+Diffusion+of+Ring+Polymer+Molecules+Revealed+by+a+Single-Molecule+Study.">Multimode Diffusion of Ring Polymer Molecules Revealed by a Single-Molecule Study.</a> Angew. Chem. Int. Ed. 49 (8), 1418-1421 (2010).</li><li>Habuchi, S., Tezuka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Topological Polymer Chemistry: Progress of Cyclic Polymers in Syntheses, Properties and Functions. , 265-290 (2013).</li><li>Fernandez, P., Bayona, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+off-line+gel-remeation+chromatography+normal-phase+liquid-chromatography+fro+the+determination+of+polycyclic+aromatic-compounds+in+environmental-samples+and+standard+reference+materials+(air+particulate+matter+and+marine+sediment).">Use of off-line gel-remeation chromatography normal-phase liquid-chromatography fro the determination of polycyclic aromatic-compounds in environmental-samples and standard reference materials (air particulate matter and marine sediment).</a> J. Chromatogr. 625 (2), 141-149 (1992).</li><li>Biesenberger, J. A., Tan, M., Duvdevan, I., Maurer, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recycle+gel+permeation+chromatography.+1.+recycle+principle+and+design.">Recycle gel permeation chromatography. 1. recycle principle and design.</a> J. Polym. Sci. Pol. Lett. 9 (5), 353 (1971).</li><li>Kusumi, A., Sako, Y., Yamamoto, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confined+lateral+diffusion+of+membrane-receptors+as+studied+by+single-particle+tracking+(nanovid+microscopy)+-+effects+of+calcium-induced+differentiation+in+cultured+epithelial-cells.">Confined lateral diffusion of membrane-receptors as studied by single-particle tracking (nanovid microscopy) - effects of calcium-induced differentiation in cultured epithelial-cells.</a> Biophys. J. 65 (5), 2021-2040 (1993).</li><li>Schutz, G. J., Schindler, H., Schmidt, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-molecule+microscopy+on+model+membranes+reveals+anomalous+diffusion.">Single-molecule microscopy on model membranes reveals anomalous diffusion.</a> Biophys. J. 73 (2), 1073-1080 (1997).</li><li>Vrljic, M., Nishimura, S. Y., Brasselet, S., Moerner, W. E., McConnell, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+diffusion+of+individual+class+II+MHC+membrane+proteins+in+cells.">Translational diffusion of individual class II MHC membrane proteins in cells.</a> Biophys. J. 83 (5), 2681-2692 (2002).</li><li>Margineanu, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photophysics+of+a+water-soluble+rylene+dye:+Comparison+with+other+fluorescent+molecules+for+biological+applications.">Photophysics of a water-soluble rylene dye: Comparison with other fluorescent molecules for biological applications.</a> J. Phys. Chem. B. 108 (32), 12242-12251 (2004).</li><li>Tezuka, Y., Oike, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-assembly+and+covalent+fixation+for+topological+polymer+chemistry.">Self-assembly and covalent fixation for topological polymer chemistry.</a> Macromol. Rapid Commun. 22 (13), 1017-1029 (2001).</li><li>Deres, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Origin+of+Heterogeneity+of+Polymer+Dynamics+near+the+Glass+Temperature+As+Probed+by+Defocused+Imaging.">The Origin of Heterogeneity of Polymer Dynamics near the Glass Temperature As Probed by Defocused Imaging.</a> Macromolecules. 44 (24), 9703-9709 (2011).</li><li>Flier, B. M. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneous+Diffusion+in+Thin+Polymer+Films+As+Observed+by+High-Temperature+Single-Molecule+Fluorescence+Microscopy.">Heterogeneous Diffusion in Thin Polymer Films As Observed by High-Temperature Single-Molecule Fluorescence Microscopy.</a> J. Am. Chem. Soc. 134 (1), 480-488 (2012).</li><li>Habuchi, S., Oba, T., Vacha, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-beam+single-molecule+defocused+fluorescence+imaging+reveals+local+anisotropic+nature+of+polymer+thin+films.">Multi-beam single-molecule defocused fluorescence imaging reveals local anisotropic nature of polymer thin films.</a> Phys. Chem. Chem. 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The authors have nothing to disclose.

Os polímeros de 4-armados e moldaram-8 foram preparados por meio do protocolo SEC-CF (figura 1), que é um passo crítico para a síntese. 12,24 monofuncional e poli bifuncional linear (THF) com N s grupos terminais foram -phenylpiperidinium sintetizado de acordo com o procedimento anterior. 11 a permuta iónica foi realizada por reprecipitação de uma solução de acetona de um precursor de polímero com contra-aniões triflato, para uma solução aquosa contendo uma qu...

Cyclic polymers are unique in that they do not have chain ends. They often exhibit unusual behaviors which is distinct from their linear counterpart, including increased thermal stability of polymer micelles by a linear-to-cyclic conversion,1,2 and spatial organization of DNA in bacterial cells by a loop formation.3 Topological interactions between the cyclic chains are believed to be the critical factor for such unusual behaviors.4,5 Therefore, characterizing the motion and relaxation of cyclic polymers under entangled conditions has been an important research topic in polymer science for decades.6
Cyclic polymer dynamics has been investigated using both synthetic and naturally occurring molecules by means of ensemble averaged experimental methods such as nuclear magnetic resonance (NMR), light scattering, and viscosity measurements.7-9 However, these studies often suffer from impurity molecules in the samples.10 Furthermore, spatiotemporal heterogeneities of the motion of individual molecules caused by inherent structural heterogeneity of entangled polymers are often hidden behind the ensemble averaging in these studies. In order to characterize molecular level dynamics of cyclic polymers, a synthesis method that provides high purity cyclic polymers and an experimental and analysis methods that allow for quantitative characterization of molecular motion at the single molecule level have to be developed. Here, we will show a method to synthesize high-purity cyclic and dicyclic poly(THF)s that incorporate a fluorophore unit using an electrostatic self-assembly and covalent fixation (ESA-CF) process11-13 and a method to analyze the motion of the individual fluorophore-incorporated polymer chains using a combination of mean-squared displacement (MSD) and cumulative distribution function (CDF) analyses.
A proper data processing has been shown to be essential for the accurate characterization of the diffusive motion. With an adequate MSD and CDF analyses, a multiple-mode diffusion of the cyclic and dicyclic polymers in the melt and semi-dilute solution of the linear polymer chains has been revealed,14-16 suggesting the significant effects of the topological states of the polymers on the diffusive motion of the chains under entangled conditions.17 While the experimental and analytical approaches to characterize the motion of the cyclic polymers are described in this protocol, the same method can be used to quantitatively characterize the diffusive motion in many other heterogeneous systems. The approach would be especially suitable when multiple diffusion components existing in the samples are to be analyzed.

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			<td height="21" style="height:21px;width:320px;">Materials</td>
			<td style="width:240px;"></td>
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			<td style="width:167px;"></td>
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			<td height="21" style="height:21px;width:320px;">THF</td>
			<td style="width:240px;">Godo</td>
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			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Wakosil C-300</td>
			<td style="width:240px;">Wako Pure Chemical Industries</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Acetone</td>
			<td style="width:240px;">Godo</td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Toluene</td>
			<td style="width:240px;">Godo</td>
			<td style="width:119px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">n-Hexane</td>
			<td style="width:240px;">Godo</td>
			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:320px;">CHCl3</td>
			<td style="width:240px;">Kanto Chemical</td>
			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:320px;">Bio-Beads S-X1</td>
			<td style="width:240px;">Bio-Rad</td>
			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:320px;">Methyl triflate</td>
			<td style="width:240px;">Nacalai Tesque</td>
			<td style="width:119px;"></td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Triflic anhydride</td>
			<td style="width:240px;">Nacalai Tesque</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Potassium Hydroxide</td>
			<td style="width:240px;">Wako Pure Chemical Industries</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Ethanol</td>
			<td style="width:240px;">Wako Pure Chemical Industries</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Poly(tetrahydrofuran)</td>
			<td style="width:240px;">Aldrich</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Chloroform</td>
			<td style="width:240px;">Wako Pure Chemical Industries</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Immersion oil</td>
			<td style="width:240px;">Cargille</td>
			<td style="width:119px;">Type 37 / Type A</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;"></td>
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			<td height="21" style="height:21px;width:320px;"></td>
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			<td height="21" style="height:21px;width:320px;">Equipment</td>
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			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">2-Neck 100-mL round-bottom flask</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Flask</td>
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			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:320px;">Beaker</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:320px;">Funnel</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Filter paper</td>
			<td style="width:240px;">Whatman</td>
			<td style="width:119px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Reflux condenser</td>
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			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:320px;">Syringe</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:320px;">Water bath</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Magnetic stirrer</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Rotary evaporator</td>
			<td style="width:240px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Microscope cover slips (24 x 24 mm, No. 1)</td>
			<td style="width:240px;">Matsunami Glass</td>
			<td style="width:119px;">CO22241</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Staining jar</td>
			<td style="width:240px;">AS ONE Corporation</td>
			<td style="width:119px;">1-7934-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Ultrasonic cleaner</td>
			<td style="width:240px;">VWR International&#160;</td>
			<td style="width:119px;">142-0047</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Inverted microscope</td>
			<td style="width:240px;">Olympus</td>
			<td style="width:119px;">IX71</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Ar-Kr ion laser</td>
			<td style="width:240px;">Coherent</td>
			<td style="width:119px;">Innova 70C</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Berek compensator</td>
			<td style="width:240px;">Newport</td>
			<td align="right" style="width:119px;">5540</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Excitation filter</td>
			<td style="width:240px;">Semrock</td>
			<td style="width:119px;">LL01-488-12.5</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Dichloric mirror</td>
			<td style="width:240px;">Omega optical</td>
			<td style="width:119px;">500DRLP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Emission filter</td>
			<td style="width:240px;">Semrock</td>
			<td style="width:119px;">BLP01-488R-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Lens and mirror</td>
			<td style="width:240px;">Thorlabs</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">EM-CCD camera</td>
			<td style="width:240px;">Andor Technology</td>
			<td style="width:119px;">iXon</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Objective lens (x100, N.A. = 1.3)</td>
			<td style="width:240px;">Olympus</td>
			<td style="width:119px;">UPLFLN 100XOP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Objective heater</td>
			<td style="width:240px;">Bioptechs</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:320px;">Preparative GPC</td>
			<td style="width:240px;">Japan Analytical Industry</td>
			<td style="width:119px;">LC-908</td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis of cyclic polymers and characterization of their diffusive motion in the melt state at the single molecule level

We demonstrate a method for the synthesis of cyclic polymers and a protocol for characterizing their diffusive motion in a melt state at the single molecule level. An electrostatic self-assembly and covalent fixation (ESA-CF) process is used for the synthesis of the cyclic poly(tetrahydrofuran) (poly(THF)). The diffusive motion of individual cyclic polymer chains in a melt state is visualized using single molecule fluorescence imaging by incorporating a fluorophore unit in the cyclic chains. The diffusive motion of the chains is quantitatively characterized by means of a combination of mean-squared displacement (MSD) analysis and a cumulative distribution function (CDF) analysis. The cyclic polymer exhibits multiple-mode diffusion which is distinct from its linear counterpart. The results demonstrate that the diffusional heterogeneity of polymers that is often hidden behind ensemble averaging can be revealed by the efficient synthesis of the cyclic polymers using the ESA-CF process and the quantitative analysis of the diffusive motion at the single molecule level using the MSD and CDF analyses.

O perylene diimida-incorporada estrelas 4 armado e em forma de 8 poli dicíclico (THF) s foram sintetizados utilizando a auto-montagem eletrostática e fixação covalente (ESA-CF) processo (Figura 1, Figura 2). Imagens de fluorescência de molécula única de lapso de tempo foram medidos para o 4-armada (Figura 3a) e 8 em forma de polímeros (Figura 3B). As imagens de lapso de tempo de fluorescência (Figura 3)<...

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Synthesis of Cyclic Polymers and Characterization of Their Diffusive Motion in the Melt State at the Single Molecule Level

Um protocolo para a síntese e caracterização de movimento difusivo de polímeros cíclicos no único nível molécula é apresentado.

Síntese de polímeros cíclicos e Caracterização do seu movimento difusivo no estado fundido na molécula de nível único

We demonstrate a method for the synthesis of cyclic polymers and a protocol for characterizing their diffusive motion in a melt state at the single ...

synthesis-cyclic-polymers-characterization-their-diffusive-motion

Research

JoVE Journal

Chemistry

7.8K visualizações.  King Abdullah University of Science and Technology (KAUST).  Um protocolo para a síntese e caracterização de movimento difusivo de polímeros cíclicos no único nível molécula é apresentado. 

Vídeo: Synthesis of Cyclic Polymers and Characterization of Their Diffusive Motion in the Melt State at the Single Molecule Level

Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria

Lipopolysaccharide (LPS) is the major cell surface molecule of gram-negative bacteria, deposited on the outer leaflet of the outer membrane bilayer. LPS can be subdivided into three domains: the distal O-polysaccharide, a core oligosaccharide, and the lipid A domain consisting of a lipid A molecular species and 3-deoxy-D-manno-oct-2-ulosonic acid residues (Kdo). The lipid A domain is the only component essential for bacterial cell survival. Following its synthesis, lipid A is chemically modified in response to environmental stresses such as pH or temperature, to promote resistance to antibiotic compounds, and to evade recognition by mediators of the host innate immune response. The following protocol details the small- and large-scale isolation of lipid A from gram-negative bacteria. Isolated material is then chemically characterized by thin layer chromatography (TLC) or mass-spectrometry (MS). In addition to matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) MS, we also describe tandem MS protocols for analyzing lipid A molecular species using electrospray ionization (ESI) coupled to collision induced dissociation (CID) and newly employed ultraviolet photodissociation (UVPD) methods. Our MS protocols allow for unequivocal determination of chemical structure, paramount to characterization of lipid A molecules that contain unique or novel chemical modifications. We also describe the radioisotopic labeling, and subsequent isolation, of lipid A from bacterial cells for analysis by TLC. Relative to MS-based protocols, TLC provides a more economical and rapid characterization method, but cannot be used to unambiguously assign lipid A chemical structures without the use of standards of known chemical structure. Over the last two decades isolation and characterization of lipid A has led to numerous exciting discoveries that have improved our understanding of the physiology of gram-negative bacteria, mechanisms of antibiotic resistance, the human innate immune response, and have provided many new targets in the development of antibacterial compounds.

Isolation and characterization of the lipid A domain of lipopolysaccharide (LPS) from gram-negative bacteria provides insight into cell surface based mechanisms of antibiotic resistance, bacterial survival and fitness, and how chemically diverse lipid A molecular species differentially modulate host innate immune responses.

Isolamento e Caracterização química do lípido A de bactérias Gram-negativas

Lipopolysaccharide (LPS) is the major cell surface molecule of gram-negative bacteria, deposited on the outer leaflet of the outer membrane bilayer. LPS ...

Química

Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid phase by using combinatorial chemistry. One of the most well studied classes of peptidomimetics is peptoids. Peptoids are easy to synthesize and have been shown to be proteolysis-resistant and cell-permeable. Over the past decade, many useful protein ligands have been identified through screening of peptoid libraries. However, most of the ligands identified from peptoid libraries do not display high affinity, with rare exceptions. This may be due, in part, to the lack of chiral centers and conformational constraints in peptoid molecules. Recently, we described a new synthetic route to access peptide tertiary amides (PTAs). PTAs are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. With side chains on both &#945;-carbon and main chain nitrogen atoms, the conformation of these molecules are greatly constrained by sterical hindrance and allylic 1,3 strain. (Figure 1) Our study suggests that these PTA molecules are highly structured in solution and can be used to identify protein ligands. We believe that these molecules can be a future source of high-affinity protein ligands. Here we describe the synthetic method combining the power of both split-and-pool and sub-monomer strategies to synthesize a sample one-bead one-compound (OBOC) library of PTAs.

Peptide tertiary amides (PTAs) are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. Here we describe a synthetic method which combines&#160;both split-and-pool and sub-monomer strategies to synthesize a one-bead one-compound library of PTAs.

Split-and-piscina Síntese e Caracterização de Peptídeos Terciário Amide Biblioteca

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid ...

Amide Coupling Reaction for the Synthesis of Bispyridine-based Ligands and Their Complexation to Platinum as Dinuclear Anticancer Agents

Amide coupling reactions can be used to synthesize bispyridine-based ligands for use as bridging linkers in multinuclear platinum anticancer drugs. Isonicotinic acid, or its derivatives, are coupled to variable length diaminoalkane chains under an inert atmosphere in anhydrous DMF or DMSO with the use of a weak base, triethylamine, and a coupling agent, 1-propylphosphonic anhydride. The products precipitate from solution upon formation or can be precipitated by the addition of water. If desired, the ligands can be further purified by recrystallization from hot water. Dinuclear platinum complex synthesis using the bispyridine ligands is done in hot water using transplatin. The most informative of the chemical characterization techniques to determine the structure and gross purity of both the bispyridine ligands and the final platinum complexes is 1H NMR with particular analysis of the aromatic region of the spectra (7-9 ppm). The platinum complexes have potential application as anticancer agents and the synthesis method can be modified to produce trinuclear and other multinuclear complexes with different hydrogen bonding functionality in the bridging ligand.

This protocol describes the use of amide coupling reactions of isonicotinic acid and diaminoalkanes to form bridging ligands suitable for use in the synthesis of multinuclear platinum complexes, which combine aspects of the anticancer drugs BBR3464 and picoplatin.

Acoplamento de amida de reacção para a síntese de ligandos baseados em Bispyridine e sua complexação com platina como agentes anticancerígenos dinucleares

Amide coupling reactions can be used to synthesize bispyridine-based ligands for use as bridging linkers in multinuclear platinum anticancer drugs. ...

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the hydroxylmethylene phosphine precursor with ammonia in methanol under a nitrogen atmosphere. The N-triphosPh ligand precipitates from the solution after approximately 1 hr of reflux and can be isolated analytically pure via simple cannula filtration procedure under nitrogen. Reaction of the N-triphosPh ligand with [Ru3(CO)12] under reflux affords a deep red solution that show evolution of CO gas on ligand complexation. Orange crystals of the complex [Ru(CO)2{N(CH2PPh2)3}-&#954;3P] (2) were isolated on cooling to RT. The 31P{1H} NMR spectrum showed a characteristic single peak at lower frequency compared to the free ligand. Reaction of a toluene solution of complex 2 with oxygen resulted in the instantaneous precipitation of the carbonate complex [Ru(CO3)(CO){N(CH2PPh2)3}-&#954;3P] (3) as an air stable orange solid. Subsequent hydrogenation of 3 under 15 bar of hydrogen in a high-pressure reactor gave the dihydride complex [RuH2(CO){N(CH2PPh2)3}-&#954;3P] (4), which was fully characterized by X-ray crystallography and NMR spectroscopy. Complexes 3 and 4 are potentially useful catalyst precursors for a range of hydrogenation reactions, including biomass-derived products such as levulinic acid (LA). Complex 4 was found to cleanly react with LA in the presence of the proton source additive NH4PF6 to give [Ru(CO){N(CH2PPh2)3}-&#954;3P{CH3CO(CH2)2CO2H}-&#954;2O](PF6) (6).

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

Ruthenium phosphine complexes are widely used for homogeneous catalytic reactions such as hydrogenations. The synthesis of a series of novel tridentate ruthenium complexes bearing the N-triphos ligand N(CH2PPh2)3 is reported. Additionally, the stoichiometric reaction of a dihydride Ru&#8211;N-triphos complex with levulinic acid is described.

A Síntese, Caracterização e Reatividade de uma série de Rutênio<em&gt; N</em&gt; -triphos<sup&gt; Ph</sup&gt; Complexos

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the ...

Synthesis, Characterization, and Functionalization of Hybrid Au/CdS and Au/ZnS Core/Shell Nanoparticles

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large extinction coefficients which can be tuned across the visible spectrum. Research into the plasmonic enhancement of optical transitions has become popular, due to the possibility of altering and in some cases improving photo-absorption or emission properties of nearby chromophores such as molecular dyes or quantum dots. The electric field of the plasmon can couple with the excitation dipole of a chromophore, perturbing the electronic states involved in the transition and leading to increased absorption and emission rates. These enhancements can also be negated at close distances by energy transfer mechanism, making the spatial arrangement of the two species critical. Ultimately, enhancement of light harvesting efficiency in plasmonic solar cells could lead to thinner and, therefore, lower cost devices. The development of hybrid core/shell particles could offer a solution to this issue. The addition of a dielectric spacer between a gold nanoparticles and a chromophore is the proposed method to control the exciton plasmon coupling strength and thereby balance losses with the plasmonic gains. A detailed procedure for the coating of gold nanoparticles with CdS and ZnS semiconductor shells is presented. The nanoparticles show high uniformity with size control in both the core gold particles and shell species allowing for a more accurate investigation into the plasmonic enhancement of external chromophores.

The synthesis of uniform gold nanoparticles coated with semiconductor shells of CdS or ZnS is performed. The semiconductor coating is conducted by first depositing a silver sulfide shell and exchanging the silver cations for zinc or cadmium cations.

Síntese, caracterização e funcionalização de híbrida UA / CDS e Au / ZnS Núcleo / Shell Nanopartículas

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large ...

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of trifluoromethoxylated aromatic compounds remains a formidable challenge in organic synthesis. Conventional approaches often suffer from poor substrate scope, or require use of highly toxic, difficult-to-handle, and/or thermally labile reagents. Herein, we report a user-friendly protocol for the synthesis of methyl 4-acetamido-3-(trifluoromethoxy)benzoate using 1-trifluoromethyl-1,2-benziodoxol-3(1H)-one (Togni reagent II). Treating methyl 4-(N-hydroxyacetamido)benzoate (1a) with Togni reagent II in the presence of a catalytic amount of cesium carbonate (Cs2CO3) in chloroform at RT afforded methyl 4-(N-(trifluoromethoxy)acetamido)benzoate (2a). This intermediate was then converted to the final product methyl 4-acetamido-3-(trifluoromethoxy)benzoate (3a) in nitromethane at 120 &#176;C. This procedure is general and can be applied to the synthesis of a broad spectrum of ortho-trifluoromethoxylated aniline derivatives, which could serve as useful synthetic building blocks for the discovery and development of new pharmaceuticals, agrochemicals, and functional materials.

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

An operationally simple procedure for the synthesis of ortho-trifluoromethoxylated aniline derivatives via a two-step sequence of O-trifluoromethylation of N-aryl-N-hydroxyacetamide followed by thermally induced intramolecular OCF3-migration is reported.

Protocolo para a Síntese de Derivados de Anilina Orto-trifluorometoxilados

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of ...

Synthesis and Characterization of Supramolecular Colloids

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical properties for applications in photonics, drug delivery and coating technology. Here we present a new family of colloidal building blocks, coined supramolecular colloids, whose self-assembly is controlled through surface-functionalization with a benzene-1,3,5-tricarboxamide (BTA) derived supramolecular moiety. Such BTAs interact via directional, strong, yet reversible hydrogen-bonds with other identical BTAs. Herein, a protocol is presented that describes how to couple these BTAs to colloids and how to quantify the number of coupling sites, which determines the multivalency of the supramolecular colloids. Light scattering measurements show that the refractive index of the colloids is almost matched with that of the solvent, which strongly reduces the van der Waals forces between the colloids. Before photo-activation, the colloids remain well dispersed, as the BTAs are equipped with a photo-labile group that blocks the formation of hydrogen-bonds. Controlled deprotection with UV-light activates the short-range hydrogen-bonds between the BTAs, which triggers the colloidal self-assembly. The evolution from the dispersed state to the clustered state is monitored by confocal microscopy. These results are further quantified by image analysis with simple routines using ImageJ and Matlab. This merger of supramolecular chemistry and colloidal science offers a direct route towards light- and thermo-responsive colloidal assembly encoded in the surface-grafted monolayer.

A protocol for the synthesis and characterization of colloids coated with supramolecular moieties is described. These supramolecular colloids undergo self-assembly upon the activation of the hydrogen-bonds between the surface-anchored molecules by UV-light.

Síntese e Caracterização de Supramoleculares Colloids

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical ...

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the development of antibody drug conjugates (ADCs) and nanomedicine, fluorescent microscopy and systems chemistry. For many of these applications, specificity of the bioconjugation method used is of prime concern. The Michael addition of maleimides with cysteine(s) on the target proteins is highly selective and proceeds rapidly under mild conditions, making it one of the most popular methods for protein bioconjugation.
We demonstrate here the modification of the only surface-accessible cysteine residue on yeast cytochrome c with a ruthenium(II) bisterpyridine maleimide. The protein bioconjugation is verified by gel electrophoresis and purified by aqueous-based fast protein liquid chromatography in 27% yield of isolated protein material. Structural characterization with MALDI-TOF MS and UV-Vis is then used to verify that the bioconjugation is successful. The protocol shown here is easily applicable to other cysteine - maleimide coupling of proteins to other proteins, dyes, drugs or polymers.

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

This protocol details the important steps required for the bioconjugation of a cysteine containing protein to a maleimide, including reagent purification, reaction conditions, bioconjugate purification and bioconjugate characterization.

Síntese de Bioconjugados de proteínas<em&gt; via</em&gt; Química cisteína-maleimida

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the ...

Insights into the Interactions of Amino Acids and Peptides with Inorganic Materials Using Single-Molecule Force Spectroscopy

The interactions between proteins or peptides and inorganic materials lead to several interesting processes. For example, combining proteins with minerals leads to the formation of composite materials with unique properties. In addition, the undesirable process of biofouling is initiated by the adsorption of biomolecules, mainly proteins, on surfaces. This organic layer is an adhesion layer for bacteria and allows them to interact with the surface. Understanding the fundamental forces that govern the interactions at the organic-inorganic interface is therefore important for many areas of research and could lead to the design of new materials for optical, mechanical and biomedical applications. This paper demonstrates a single-molecule force spectroscopy technique that utilizes an AFM to measure the adhesion force between either peptides or amino acids and well-defined inorganic surfaces. This technique involves a protocol for attaching the biomolecule to the AFM tip through a covalent flexible linker and single-molecule force spectroscopy measurements by atomic force microscope. In addition, an analysis of these measurements is included.

Here we present a protocol to measure the force of interactions between a well-defined inorganic surface and either peptides or amino acids by single-molecule force spectroscopy measurements using an atomic force microscope (AFM). The information obtained from the measurement is important to better understand the peptide-inorganic material interphase.

Insights sobre as interações de aminoácidos e peptídeos com Materiais Inorgânicos Usando Single-Molecule Força Spectroscopy

The interactions between proteins or peptides and inorganic materials lead to several interesting processes. For example, combining proteins with minerals ...

Using Cyclic Voltammetry, UV-Vis-NIR, and EPR Spectroelectrochemistry to Analyze Organic Compounds

Cyclic voltammetry (CV) is a technique used in the analysis of organic compounds. When this technique is combined with electron paramagnetic resonance (EPR) or ultraviolet-visible and near-infrared (UV-Vis-NIR) spectroscopies, we obtain useful information such as electron affinity, ionization potential, band-gap energies, the type of charge carriers, and degradation information that can be used to synthesize stable organic electronic devices. In this study, we present electrochemical and spectroelectrochemical methods to analyze the processes occurring in active layers of an organic device as well as the generated charge carriers.

In this article, we describe electrochemical, electron paramagnetic resonance, and ultraviolet-visible and near-infrared spectroelectrochemical methods to analyze organic compounds for application in organic electronics.

Usando Voltametria Cíclica, UV-Vis-NIR e Espectroeletroquímica EPR para Analisar Compostos Orgânicos

Cyclic voltammetry (CV) is a technique used in the analysis of organic compounds. When this technique is combined with electron paramagnetic resonance ...