Untitled

Abstract

To investigate the spatial distribution of white egg albumin (WEA) in alginate beads, a new method based on confocal laser scanning microscopy (CLSM) was developed. In contrast to the existing CLSM methods, misleading conclusions are prevented with the application of the new method which does not allow the attenuation of the exciting and emitted light by the opaque hydrogel matrices to be disregarded. By the application of this method, the distribution of WEA in alginate beads was shown to be dependent on the amount of protein loading. At low quantities of protein, a higher protein concentration occurs in the shell layer of the alginate bead while at higher loadings a more or less homogeneous distribution is observed.

Advanced characterization of immobilized enzymes as heterogeneous biocatalysts

“…Layers thus prepared were then analyzed microscopically. Confocal laser scanning microscopy (CLSM) has significantly advanced the analysis of proteins in intact porous supports (Table 2) and researchers in protein chromatography were forerunners in this respect [62][63][64][65][66][67][68][69][70]. CLSM offers spatial resolution at a length scale well suitable for the characterization of enzyme immobilizates and it is conveniently used even by nonspecialists.…”

New insights in the spatially resolved dynamic pH measurement in macroscopic large absorbent particles by confocal laser scanning microscopy

“…When scanning an alginate bead with the z-scan mode, the intensities of the detected fluorescence signals decrease with increasing scanning depth [6]. This is also obvious from the intensity profiles presented in Fig.…”

“…Confocal laser scanning microscopes (CLSM) have been widely used for spatially resolved determination of protein concentrations inside various carrier matrices and absorbent particles of macroscopic size [1][2][3][4][5][6][7][8][9][10]. Dynamic protein absorption experiments, recently observed with confocal laser scanning microscopes, revealed unexpected protein concentration profiles, the so-called concentration rings [5,7,10].…”

Photonic Crystal Hydrogel Enhanced Plasmonic Staining for Multiplexed Protein Analysis

“…Consequently, electromagnetic field at 633 and 785 nm are likely to concentrate in hydrogel region and power of 532 nm distributes in the nanovoids. From the images of silver stained IOPHB it can be seen that most of the silver nanoparticles were locked inside the nanovoids of the inverse opal structure and most of the proteins were also distributed on the surface layer of inverse opal structure rather than inside the cross‐linked skeleton of the hydrogel . Consequently, the large fraction of 532 nm power concentrated inside the nanovoids could readily couple with LSPR band of the aggregated silver nanoparticles and enhanced the Raman signal most effectively.…”

“…At 532 nm, the energy hotspots near the hydrogel surface overlap the hotspots that formed adjacent to silver nanoparticles and in gaps between silver nanoparticle dimers. Positions of these hotspots were ideal for protein detection because the protein molecules are anchored to the hydrogel backbone and swinging inside the nanovoids . At 532 nm, silver nanoparticle decorated PhC is capable of raising the local electric field by a factor of 15 compared to the incident field.…”