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            <idno type="stamp" n="INRA">INRA - Institut national de la recherche agronomique</idno>
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                <title level="j">Cahier des Techniques de l'INRA</title>
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                <term xml:lang="en">Enzyme</term>
                <term xml:lang="en">real time imaging</term>
                <term xml:lang="en">Infrared</term>
                <term xml:lang="en">sample preparation</term>
                <term xml:lang="en">fluorescence</term>
                <term xml:lang="fr">préparation d'échantillon</term>
                <term xml:lang="fr">imagerie en temps réel</term>
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                <term xml:lang="fr">fluorescence</term>
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              <p>Various preparations of sample sections have been tested to observe enzyme localization without any previous labelling, some vegetal wall components and their evolution during the degradation. Preparation of paraffin sections has been chosen to obtain spectral and hyperspectral images in real time. Two methods of closely related and complementary imaging were set out :                 - fluorescence microscopy with excitation in deep UV allows to obtain some information about the location of enzymes by multi-scale imaging                    - FT-IR microspectroscopy with the use of microfluid cell allows to follow in real time the cell wall component change under enzyme action.</p>
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              <p>La préparation des échantillons est une étape essentielle pour leur étude par microscopie photonique et microspectroscopie. Différentes préparations de coupes d’échantillons ont été testées pour l’observation sans marquage préalable de la localisation des enzymes, des constituants des parois végétales et de leurs évolutions au cours de la dégradation. La préparation de coupes paraffine a été retenue pour l’acquisition d’images spectrales et hyperspectrales en temps réel. Deux méthodes d’imagerie complémentaires et corrélables ont été utilisées :     - la microscopie de fluorescence avec une excitation dans l’UV profond permet d’obtenir des informations sur la localisation des enzymes par imagerie multi-échelles.                   - la microspectroscopie FT-IR avec l’utilisation d’une cellule microfluidique permet le suivi en temps réel du changement de composition des parois sous l’action des enzymes.</p>
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