In Vitro Antileishmanial Activity of Achillea santolina Essential Oil against Leishmania infantum Promastigote by Methylthiazole Tetrazolium (MTT) and Trypan Blue Colorimetric Methods

Leishmaniasis causes parasitic infections, especially in developing countries. The disease has not yet been controlled because of the absence of an effective vaccine and low-cost treatment. Achillea santolina essential oil (ASEO) might control the disease as it has antimicrobial properties. This study investigated the in vitro antileishmanial activity of ASEO against Leishmania infantum promastigote using the methylthiazole tetrazolium (MTT) and trypan blue colorimetric methods. The standard strain of L. infantum (MCAN/IR/96/LON49) promastigotes was prepared and cultured in a 96-well Novy-MacNeal-Nicolle (NNN) medium. The effects of different concentrations of saline, ASEO, and glucantime (10, 50, 100, 200, 500, and 1000 mg/mL) were examined in 24-, 48-, and 72-hour intervals using the MTT and trypan blue test methods.The use of ASEO reduced viability in all concentrations compared to the control group in times of 48 (p<0.05) and 72 h (p<0.05). Treatment with glucantime and ASEO had similar efficiency with the concentration of 1000 mL/mg in both methods after 72 h. The results showed that viability was significantly lower in the ASEO group with increases in time using both methods (p<0.05). Cohen’s Kappa coefficient showed a significant agreement between the obtained results for the two methods (Kappa=0.856; p<0.001).In sum, the results showed in vitro antileishmanial activity of ASEO, but more clinical studies are needed to confirm the efficiency. ASEO can be used as an agent and/or in combination with synthetic agents for the treatment of leishmaniasis disease.

Label-free cell based impedance measurements of ZnO nanoparticles-human lung cell interaction: a comparison with MTT, NR, Trypanblue and cloning efficiency assays

Background: There is a huge body of literature data on ZnOnanoparticles (ZnO NPs) toxicity. However, the reported results are seen to be increasingly discrepant, and deep comprehension of the ZnO NPs behaviour in relation to the different experimental conditions is still lacking. A recent literature overview emphasizes the screening of the ZnO NPs toxicity with more than one assay, checking the experimental reproducibility also versus time, which is a key factor for the robustness of the results. In this paper we compared high-throughput real-time measurements through Electric Cell-substrate Impedance-Sensing (ECIS®) with endpoint measurements of multiple independent assays.
Results: ECIS-measurements were compared with traditional cytotoxicity tests such as MTT, Neutral red, Trypan blue, and cloning efficiency assays. ECIS could follow the cell behavior continuously and noninvasively for days, so that certain long-term characteristics of cell proliferation under treatment with ZnO NPs were accessible. This was particularly important in the case of pro-mitogenic activity exerted by low-dose ZnO NPs, an effect not revealed by endpoint independent assays.
This result opens new worrisome questions about the potential mitogenic activity exerted by ZnO NPs, or more generally by NPs, on transformed cells. Of importance, impedance curve trends (morphology) allowed to discriminate between different cell death mechanisms (apoptosis vs autophagy) in the absence of specific reagents, as confirmed by cell structural and functional studies by high-resolution microscopy. This could be advantageous in terms of costs and time spent. ZnO NPs-exposed A549 cells showed an unusual pattern of actin and tubulin distribution which might trigger mitotic aberrations leading to genomic instability.
Conclusions: ZnO NPs toxicity can be determined not only by the intrinsic NPs characteristics, but also by the external conditions like the experimental setting, and this could account for discrepant data from different assays. ECIS has the potential to recapitulate the needs required in the evaluation of nanomaterials by contributing to the reliability of cytotoxicity tests. Moreover, it can overcome some false results and discrepancies in the results obtained by endpoint measurements. Finally, we strongly recommend the comparison of cytotoxicity tests (ECIS, MTT, Trypan Blue, Cloning efficiency) with the ultrastructural cell pathology studies.
Keywords: Confocal microscopy; Cytotoxicity tests; Electric cell-substrate impedance; Electron microscopy; Nanomaterials; Zinc oxide nanoparticles.

Masked comparison of trypan blue stain and potassium hydroxide with calcofluor white stain in the microscopic examination of corneal scrapings for the diagnosis of microbial keratitis

Purpose: To evaluate the efficacy of trypan blue in direct microscopic examination of corneal scrapings in the diagnosis of non-viral microbial keratitis.
Methods: In a prospective, interventional, masked study, 82 consecutive patients were investigated. Direct microscopic examination of the corneal scrapings involved three smears stained with potassium hydroxide with calcofluor white (KOH + CFW), Gram stain (not analyzed), and trypan blue stain and culture for bacteria, fungus, and Acanthamoeba. While KOH + CFW stained slides were examined under a fluorescence microscope, trypan blue-stained slides were examined by two microbiologists (masked to KOH + CFW and culture results) under normal light

Purpose: In the present clinical study, it was aimed to investigate the possible effects of Trypan blue (TB) use on the corneal endothelium during cataract surgery in eyes with pseudoexfoliation syndrome (PEX) during a three-month follow-up period using the contralateral eye control design.
Methods: This prospective, randomised controlled, individual cohort study included 92 eyes of 46 patients with bilateral PEX and cataracts. While 1% TB was applied to one eye of the patients before capsulorhexis (study group), it was not applied to the other eye (control group). Both groups were compared preoperatively and postoperatively in terms of endothelial cell density (ECD), endothelial cell loss (%), pleomorphism, polymegathism and central corneal thickness (CCT) using specular microscopy.

microscopy. Thirty samples were reexamined for interobserver and intraobserver variability.

The effects of trypan blue use on the corneal endothelium during cataract surgery in patients with pseudoexfoliation syndrome (PEX)

Purpose: In the present clinical study, it was aimed to investigate the possible effects of Trypan blue (TB) use on the corneal endothelium during cataract surgery in eyes with pseudoexfoliation syndrome (PEX) during a three-month follow-up period using the contralateral eye control design.
Methods: This prospective, randomised controlled, individual cohort study included 92 eyes of 46 patients with bilateral PEX and cataracts. While 1% TB was applied to one eye of the patients before capsulorhexis (study group), it was not applied to the other eye (control group). Both groups were compared preoperatively and postoperatively in terms of endothelial cell density (ECD), endothelial cell loss (%), pleomorphism, polymegathism and central corneal thickness (CCT) using specular microscopy.

Trypan Blue – Adapting a Dye Used for Labelling Dead Cells to Visualize Pinocytosis in Viable Cells.

Background/aims: Trypan blue is routinely used in cell culture experiments to distinguish between dead cells, which are labelled by trypan blue, and viable cells, which are apparently free of any staining. The assumption that trypan blue labelling is restricted to dead cells derives from the observation that rupture of the plasma membrane correlates with intense trypan blue staining.

Trypan Blue

20-abx082476 Abbexa
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Trypan Blue

40140044-1 Glycomatrix 25 g 22.88 EUR

Trypan Blue

40140044-2 Glycomatrix 100 g 66.8 EUR

Trypan Blue

58026 Sisco Laboratories 25 Gms 7.95 EUR

Trypan Blue

abx082476-100l Abbexa 100 µl 137.5 EUR

Trypan Blue

abx082476-1ml Abbexa 1 ml Ask for price

Trypan Blue

abx082476-200l Abbexa 200 µl 200 EUR

Trypan blue

TT1140 Bio Basic 10g 78.79 EUR

Trypan Blue Trypan Blue Solution, 0.4% Solution in PBS.

TBL02-100ML Caisson Labs 100 ml 98.4 EUR

Trypan Blue (0.4%)

1209-10 Biovision each 157.2 EUR

Trypan Blue (C.I. 23850)

GT3970 Glentham Life Sciences 25g 72.46 EUR

Trypan Blue (C.I. 23850)

GT3970-100 Glentham Life Sciences 100 79.1 EUR

Trypan Blue (C.I. 23850)

GT3970-100G Glentham Life Sciences 100 g 132 EUR

Trypan Blue (C.I. 23850)

GT3970-25 Glentham Life Sciences 25 31.7 EUR

Trypan Blue (C.I. 23850)

GT3970-25G Glentham Life Sciences 25 g 74.4 EUR

Trypan Blue (C.I. 23850)

GT0822-GT0822100ml Glentham Life Sciences GT0822-100ml 47.4 EUR

Trypan Blue Assay Kit

AR1175 BosterBio 1kit (for 1000 assays) 78 EUR

Trypan Blue 0.4% Solution

25561-250 Polysciences Europe GmbH 250ml 56 EUR

Trypan Blue 0.4% Solution

40140051-1 Glycomatrix 100 mL 12.81 EUR

Trypan Blue 0.4% Solution

40140051-2 Glycomatrix 15 mL 25.8 EUR

Trypan Blue 0.4% Solution

12788 Sisco Laboratories 50 ml 3 EUR

0.4% Trypan Blue Solution

K1183-100 ApexBio 100ml 64 EUR

0.4% Trypan Blue Solution

K1183-50 ApexBio 50ml 40 EUR

0.4% Trypan Blue Solution

PB180423-100mL Elabscience Biotech 100 mL 20 EUR

0.4% Trypan Blue Solution

PB180423 Elabscience Biotech 100mL 20 EUR

0.4% Trypan Blue solution

MBS2567254-100mL MyBiosource 100mL 95 EUR
However, decades ago, trypan blue has been used to trace fluid uptake by viable macrophage-like cells in animals. These studies contributed to the concept of the reticuloendothelial system in vertebrates. Trypan blue itself does not show a fluorescence signal, but trypan blue-labelled proteins do. Therefore, intracellular localization of trypan blue-labelled proteins could give a clue to the entrance pathway of the dye in viable cells.

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