Inadvertent trypan blue staining of posterior capsule during cataract surgery associated with Argentinian flag event

Trypan blue is common in visualizing the anterior capsule during cataract surgery. Inadvertent staining of the posterior capsule during phacoemulsification is a rare complication and there are few reports in the literature. The proposed mechanism of posterior capsule staining in previous reports includes a compromised zonular apparatus or iris retractors facilitating the posterior flow of trypan blue. We report the first case of trypan blue staining of the posterior capsule associated with the “Argentinian flag” sign. In our case, the “Argentinian flag” allowed the trypan blue to seep between the posterior capsule and the lens, staining the anterior surface of the posterior capsule

Trypan Blue Dye Enters Viable Cells Incubated with the Pore-Forming Toxin HlyII of Bacillus cereus

Trypan blue is a dye that has been widely used for selective staining of dead tissues or cells. Here, we show that the pore-forming toxin HlyII of Bacillus cereus allows trypan blue staining of macrophage cells, despite the cells remaining viable and metabolically active. These findings suggest that the dye enters viable cells through the pores. To our knowledge, this is the first demonstration that trypan blue may enter viable cells. Consequently, the use of trypan blue staining as a marker of vital status should be interpreted with caution. The blue coloration does not necessarily indicate cell lysis, but may rather indicate pore formation in the cell membranes and more generally increased membrane permeability

Electron microscope studies on the relationship between mitochondria and inclusion body in spheroid alveolar epithelial cell after vital staining with acidic and basic dyes

Electron microscope observations were conducted on the relationship between mitochondria and inclusion body in mice spheroid alveolar epithelial cells after injection of trypan blue, an acidic dye and Alcian blue 8GS, a basic dye, by vital staining procedures. When both dyes were injected, the mitochondria of the spheroid alveolar epithelial cell became degenerated; however, in injection of only trypan blue, the cristae showed an increase in electron density. In injection on only Alcian blue 8GS, the cristae showed negative contrast. In most cases the trypan blue particles did not enter into mitochondria, whereas particles of Alcian blue 8GS sometimes entered into the mitochondria. When trypan blue particles entered mitochondria, deposits were not evident in the inclusion body, whereas when Alcian blue particles entered mitochondria deposits were seen in the inclusion body. In both of these cases only a few inclusion bodies were formed so that only traces or no inclusion bodies with vacuolar appearance were observed. From these findings it is suggested that mitochondria maybe convert to inclusion bodies.</p

A comparison of methodologies for the staining and quantification of intracellular components of Arbuscular Mychorrizal (AM) fungi in the root cortex of two varieties of winter wheat

© 2019 The Authors. The definitive peer reviewed, edited version of this article is published in Access Microbiology, https://doi.org/10.1099/acmi.0.000083. This is an open-access article distributed under the terms of the Creative Commons Attribution License.Arbuscular Mychorrizal (AM) fungi are one of the most common fungal organisms to exist in symbiosis with terrestrial plants facilitating the growth and maintenance of arable crops. Wheat has been studied extensively for AM fungal symbiosis using the carcinogen trypan blue as the identifying stain for fungal components, namely arbuscles, vesicles and hyphal structures. The present study uses Sheaffer® blue ink with a lower risk as an alternative to this carcinogenic stain. Justification for this is determined by stained wheat root sections (n = 120), with statistically significant increases in the observed abundance of intracellular root cortical fungal structures stained with Sheaffer® blue ink compared to trypan blue for both Zulu (P = 0.003) and Siskin (P = 0.0003) varieties of winter wheat. This new alternative combines an improved quantification of intracellular fungal components with a lower hazard risk at a lower cost.Peer reviewe

Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) settingg

Aim: To compare our parameters as regards: i) cell count via two different automated cell count techniques, and ii) viability via automated trypan blue exclusion and 7-aminoactinomycin D (7-AAD) staining. Method: We used the trypan blue exclusion technique and an automated cell counter and for viability testing, and the trypan blue exclusion technique and the 7-AAD evaluation by flow cytometry. The trypan blue exclusion and the radio frequency techniques were used for automated cell counting. Flow cytometric analysis was performed by evaluating the yielded cellular products for 7-AAD uptake during the cell count of CD34+ cells. Results: The mean values for cell count were estimated as 3.44±1.22x106/ml (range, 2.48-5.71x106/ml) and 4.14±1.94x106/ml (range, 1.77-7.43x106/ml) for the trypan blue exclusion and radio frequency techniques, respectively. Additionally, the mean values for viability analyses via the automated trypan blue exclusion and 7-AAD were 93.38±6.09% (range, 79.00-98.00%) and 99.49±0.60% (range, 98.40-100.00%), respectively. Conclusions: Our study has responded to two fundamental questions: whether the results of both of the automated techniques for cell count correspond with each other, and whether the results of the automated viability assessment conform those of the 7-AAD technique during the manufacturing processes of cellular therapy products intended for clinical use. Even though we have the opportunity to use the hemocytometer in our laboratory setting, the automated trypan blue exclusion technique gives cell count results in concordance within the range of the expectations of our Quality Management System (QMS)

Pembersihan Isi Sel Akar dan Jenis Warna Tinta untuk Deteksi Cendawan Mikoriza Arbuskula: Clearing of Root Cell Content and Types of Ink Stain for Arbuscular Mycorrhizal Fungal Detection

Arbuscular mycorrhizal (AM) fungi form mutualistic symbiosis with root of host plant. Staining technique to detect AM fungi usually used hazardous chemical. The ink stain and vinegar were used as an alternative technique to replace trypan blue and lactic acid in root staining method. This study aimed to determine time for clearing root cell contents and ink stain type to visualize the best AM fungal structures within the root observed under light microscope. Pueraria phaseoloides var. javanica roots colonized by AM fungi were cut into 1 cm long, cleared in KOH solution and stained.&nbsp; Four clearing time were done vis 5, 10, 15 and 20 minutes, and four stains were used namely Shaeffer black ink, Parker Quink blue ink, blue stamp ink, and trypan blue. Twenty stained roots were taken randomly from each tratment, and observed. Root clearing process for 20 minutes showed the best result. Only Shaeffer black ink and trypan blue produced clear structure of external hyphae, internal hyphae, vesicles and arbuscules. Arbuscular structure stained only by Shaeffer black ink and trypan blue. This indicated that Shaeffer black ink could be used as an alternative stain to detect AM fungi within the root of host plantArbuscular mycorrhizal (AM) fungi form mutualistic symbiosis with root of host plant. Staining technique to detect AM fungi usually used hazardous chemical. The ink stain and vinegar were used as an alternative technique to replace trypan blue and lactic acid in root staining method. This study aimed to determine time for clearing root cell contents and ink stain type to visualize the best AM fungal structures within the root observed under light microscope. Pueraria phaseoloides var. javanica roots colonized by AM fungi were cut into 1 cm long, cleared in KOH solution and stained.&nbsp; Four clearing time were done vis 5, 10, 15 and 20 minutes, and four stains were used namely Shaeffer black ink, Parker Quink blue ink, blue stamp ink, and trypan blue. Twenty stained roots were taken randomly from each tratment, and observed. Root clearing process for 20 minutes showed the best result. Only Shaeffer black ink and trypan blue produced clear structure of external hyphae, internal hyphae, vesicles and arbuscules. Arbuscular structure stained only by Shaeffer black ink and trypan blue. This indicated that Shaeffer black ink could be used as an alternative stain to detect AM fungi within the root of host plan

Bonghan Ducts as Possible Pathways for Cancer Metastasis

Objective: The present study has been designed to find a possible new route for the metastasis of cancer cells on the fascia surrounding tumor tissue using a novel technique of trypan blue staining. Materials and Methods: Tumor tissues were grown in the skin of nude mice after subcutaneous inoculation with human lung cancer cells. Trypan blue was recently identified as a dye with specificity for Bonghan ducts (BHDs) and not other tissues, such as blood or lymph vessels or nerves. Results: We demonstrate that the trypan blue staining technique allows the first visualization of BHDs which are connected to tumor tissues

Bonghan Ducts as Possible Pathways for Cancer Metastasis

Objective: The present study has been designed to find a possible new route for the metastasis of cancer cells on the fascia surrounding tumor tissue using a novel technique of trypan blue staining. Materials and Methods: Tumor tissues were grown in the skin of nude mice after subcutaneous inoculation with human lung cancer cells. Trypan blue was recently identified as a dye with specificity for Bonghan ducts (BHDs) and not other tissues, such as blood or lymph vessels or nerves. Results: We demonstrate that the trypan blue staining technique allows the first visualization of BHDs which are connected to tumor tissues

Photochemical degradation of trypan blue

Purpose To investigate the photochemical degradation of trypan blue (TB) and to identify decomposition products. Methods Defined solution samples of TB and a mixture with lutein/zeaxanthin were exposed to blue light. Thermal degradation processes were ruled out using controls not subjected to irradiation. All samples were analyzed using optical microscopy, UVNis spectroscopy, matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and nuclear magnetic resonance (NMR) spectrometry. Degradation kinetics were determined based on changes in absorbance; intermediates were identified by analyzing mass differences of characteristic fragment ion peaks within the fragmentation patterns, and assignments were verified by NMR. Results TB demonstrated a photochemical degradation, which can be triggered by lutein/zeaxanthin. Intermediates vary depending on the presence of lutein/zeaxanthin. The self-sensitized photodegradation of TB occurs under generation of dimethyl sulfate and presumed formation of phenol. In contrast, within the presence of lutein/zeaxanthin the decomposition of TB indicates the formation of methoxyamine and sulfonyl arin. Thermal degradation processes were not observed. Conclusions TB demonstrated a photodegradation that may be triggered by lutein/zeaxanthin and results in the formation of cytotoxic decomposition products. Our findings contribute to understand degradation mechanisms of TB and may elucidate previous clinical and experimental observations of cellular toxicity after TB application