Novel application of aniline blue

Abstract

The present study shows that aniline blue can be used as a fluorescent stain for glycogen. The dye is also helpful in tracing pathological and autolytic changes in lysosomes, mitochondria, erythrocytes and nuclei, and it can also be used for demonstrating bacteria in tissue sections and smears. The techniques used are simple, rapid and inexpensive. Spectrophotometric studies on aniline blue solutions have shown that aniline blue fluorescence was enhanced by the addition of certain proteins, or of glycogen to the dye solution. In case of albumen which has the maximum effect, enhancement is dependent upon the albumen-dye ratio. The mechanism of staining is mainly due to self quenching, but there is also an evidence of the presence of hydrophobic reaction.

Prediction of intracellular storage polymers using quantitative image analysis in enhanced biological phosphorus removal systems

“…Upon comparison between the quantitative image analysis results, for the acquired fluorescent images of the two studies, and the sample glycogen analytical concentration obtained via HPLC, better results were obtained using the 2% alkaline solution [31]. This agreed with the findings of Shennawy et al [31]. Thus, the methodology used for biomass suspension was as follows.…”

“…Intracellular glycogen storage was observed using Aniline blue staining. Although aniline blue is generally used to detect glycogen in tissue sections [31], in activated sludge the application of aniline blue staining procedures for glycogen detection has been questioned and not frequently used due to inconsistent results [32]. In order to try to overcome the limitations of the existing methodologies, two comparative studies (data not shown) were performed using 2% acid solution (acetic acid with pH at 3.5) and alternatively 2% alkaline solution (phosphate buffer with pH at 7.2) during 20 min reaction [31] with biomass suspension.…”

“…Although aniline blue is generally used to detect glycogen in tissue sections [31], in activated sludge the application of aniline blue staining procedures for glycogen detection has been questioned and not frequently used due to inconsistent results [32]. In order to try to overcome the limitations of the existing methodologies, two comparative studies (data not shown) were performed using 2% acid solution (acetic acid with pH at 3.5) and alternatively 2% alkaline solution (phosphate buffer with pH at 7.2) during 20 min reaction [31] with biomass suspension. Upon comparison between the quantitative image analysis results, for the acquired fluorescent images of the two studies, and the sample glycogen analytical concentration obtained via HPLC, better results were obtained using the 2% alkaline solution [31].…”

“…In order to try to overcome the limitations of the existing methodologies, two comparative studies (data not shown) were performed using 2% acid solution (acetic acid with pH at 3.5) and alternatively 2% alkaline solution (phosphate buffer with pH at 7.2) during 20 min reaction [31] with biomass suspension. Upon comparison between the quantitative image analysis results, for the acquired fluorescent images of the two studies, and the sample glycogen analytical concentration obtained via HPLC, better results were obtained using the 2% alkaline solution [31]. This agreed with the findings of Shennawy et al [31].…”

Activated sludge characterization through microscopy: A review on quantitative image analysis and chemometric techniques

“…Combined with FISH or otherwise, fluorescent staining samples with DAPI [95], at high concentrations, and Nile blue [96] dyes, can be applied to visualize polyphosphate granules and PHA inclusions, respectively. Furthermore, Shennawy et al [97] reported the use of Aniline blue to stain glycogen inside cells. A more detailed description about the different staining techniques can be found in Serafim et al [75].…”