Dyes - Science topic

I'm having great success using Cell Tracker red with Lactobacillus. Cell Tracker Blue CMHC and Green CM-H2DCFDA aren't working. I'm doing some trouble-shooting but I'd appreciate any insights from people who have tried these dyes on bacteria.

I want to stain acute brain slices with calcium orange and DAPI, but I am not sure whether these two kinds of dye can be added together or not.

At our lab we are trying to develop new dyes for different applications, and we need information about the stomach acidity of Galleria mellonella model.

Dear ResearchGate community, hello!

I'm going to do an MTT test on glioblastoma cells with our drugs. I plan to add 10,000 cells in 100 µl medium with FBS per well to a 96-well plate, incubate for 24 hours and then add 50 µl of drugs. Incubate the cells with treatment for 24 hours and add MTT dye.

Tell me, please, should I remove the medium before adding the drugs (they are diluted in DMEM without FBS) and should I remove the drugs before adding MTT dye?

Thank you in advance!

I am searching for a fluorescence far red dye to stain bacteria for live Microscopic Imaging. PSVUE is one of NIR (Near Infrared Dye) which stains anionic lipids but it is not compatible with PBS buffer which contains anionic phosphates. Do anyone have used this dye with RPMI 1640+ FBS culture medium? Is it compatible with it or not?

Also have anybody used  other dye named DRAQ5 for lie imging for bacteria?

I want to stain bull sperm cells (dead/alive) with Hoechst 33342 (10 mg/mL in H2O) and don't know how to do it properly. I will be grateful if you could help me. Best regards and stay healthy.

I am having trouble distinguishing an unhatched/dead C. elegans embryo from a remaining egg-shell of a hatched embryo in a bright-field image in a high-throughput fitness assay.

I was wondering if one could discriminate between an unhatched/dead embryo and the remaining chitin shell by adding some dye to it. DNA dyes (Hoechst 33342, 33258, Sytox green) are not penetrating the embryo. Chitin dyes (Calcofluor white, congo red) are just staining the chitin shell of a hatched and an unhatched embryo in a similar fashion.

I would be happy if someone would provide an idea.

I have a recipe of loading dye (10 mM EDTA, 1X TBE, pinch of Xylene cynol and Orange G dye which is made upto 10 ml with 100% Formamide) but somehow the bands of the nucleotide product (enzymatically degraded DNA) are not distinct and well defined or fuzzy. I use 1 X TBE buffer (freshly prepared), the volt used to run the gel is 1500-2000 Volt or 25-40 W. Could you please suggest the recipe of loading dye or other factors that could give crisp bands for publication purpose. Any advice would be highly appreciated.

Thank you in advance

Prem

I am planning to study adsorption efficiency of dye at different pH. As we know, some dyes would change colour at different pH. Therefore, is it necessary to plot different calibration curve at each pH? 7 different pH with 4 different dye concentrations would mean 28 solutions that have to be made. Is this a common approach?

for photo phenton reaction, dye concentration 10ppm, catalyst 100mg/L.

Explain anyone after completely removed dye from contaminated water, what about catalyst whether catalyst also removed or not

I am studying the anti-tumor effect of Hoechst33342 dye on A-375 cells. However, when I added 75uM of the dye to the plated cells, the dye seem to precipitate and I get close to 100% cell death. Is this normal? If not what can I do to avoid this precipitation? (the stock Hoechst33342 solution is prepared in distilled autoclave water)

Relative molar composition of

1.0 TEOS : 0.2 CTACl : 0.0026–0.017 LS277

dye : 10.4 TEA : 142 H2O was used.

I am trying to do some fluorescent microscopy on E.coli and P.aeruginosa cells after 24h treatments with compounds. I am using propidium iodide (molecular probes) and SYTO 9 (Thermo) in the following way:

1. Add equal volumes of each dye to 100ul of 20% glycerol

2. Add 2ul of the dye to mixture to samples in microtiter plate

3. Cover with foil and incubate in dark for 15 minutes

4. Pipette 10ul samples onto slide and view under fluorescent microscope

When I come to view my cells under the microscope, I can only see them under light microscopy and when I switch to using the fluorescent filters, I see the same cells in both filters and none of them are fluorescing green or red.

I tried just adding each stain separately and the fluorescing cells in each filter can be seen but not when I add it is a mixture of both dyes.

Could someone assist me?

Michael

I have been reading quite a few papers that deal with food dye and their effects on cells (ex. DNA damage etc). Many of them note the Acceptable Daily Intake (mg/kg) as their foundation for selecting the concentrations of the dye (µg/mL) they will use on the cells. I can't seem to find any paper that explains how they reach these numbers/do these conversions so I was wondering if anyone knew how I could find out?

I deposited a polymer dye on cotton fabric. It seems it is covalently attached. But i am confused what can be the possible mechanism for covalent interaction between the polymer and fabric?

For water treatment approach, different types of adsorbents are used. They has sufficient open hand to bind with pollutants. But, in an equilibrium study, after a certain adsorbent dose, extra doses can't remove extra quantity of effluent dye. Why?

Hello everyone,

I am looking for any dye that can be used to determine the flow of liquid in a tube. Therefore, it should not affect tissues and cells or bind to nucleic acids or proteins. Additionally, it should be easy to wash off.

Any suggestions would be greatly appreciated.

Cheers

I'm performing adsorption tests, using different adsorbent doses, its particles size is unknown.

To measure the final dye concentration I need to remove the adsorbent for it not to adsorb UV beam. I've been using centrifugation at 4000 rpm, which is the maximum velocity my centrifuge could reach, but it doesn't seem to help. I'm afraid I can't use filtration, it could contribute to the adsorption.

is it any specific dye for live cell imaging? is flow cytometer antibody Ok? or IF antibody?

Dear all,

I am wondering if you have ever experienced bleed-through using the 680and 800 infrared dye secondary antibodies from LiCor. I observed a possible bleed through from a highly expressed protein (red channel, 680nm) into the green channel (800 nm). What is your experience? How did you solve it?

Thanks

Hi,

I am trying to subclone iPSCs by plating 200-300 cells in 6 wells previously coated with geltrex. When I plate them I use 10uM of rock inhibitor and in theory I j

keep it until a nice colony is formed. I tried both E8 and stem flex media. however the day after I plate them, I got single cells but then after 2-3days they die or they remain as single cells without proliferating. I Have tried to change their media every other day as well as every day (I thought maybe the rock inhibitor at 37 degree got degraded). Any suggestion?

Why after the adsorption of methyl orange dye by my adsorbent, in addition to the color peak in 464 nm, another peak has appeared in 372nm n in theuv-vis spectrum. It should be noted that this additional peak can be seen only in low adsorbent dosage, higher color concentration and shorter conatct time!

thanks in advance for your help

Hi

Does anyone know which dye can be used for staining exosome membranes, aside from PKH67?

Thank you in advance for your help

After performing PCR, I ran electrophoresis, but on agarose the results showed some rather blurred samples. I wanted to know the cause and how to fix this situation. Please note that the chemicals and dyes are normal because the positive control shows a clear band.

I am mixing a sodium alginate gel to apply to microfluidic channels, I’d like to use a fluorescent dye to visualise where it has been deposited after application.

It would have to be oleophobic as I cannot have the dye seep into the oil and accidentally visualise oil.

Preference of a green dye too, and cheap if possible!

Hello!

I would like to synthesize dye loaded silica particles via Stöber or modified Stöber method with a particle size of 40 nm (minimum) or 100 nm (maximum). I've already searched for publications but I didn't find an appropriate synthesis where dye loading and Stöber were combined. I already tried a synthesis with Sodium Fluorescine as a dye but it didn't work well - the size was a bit too high (62) and the polydispersity around 16% and the dye intensitiy too low.

I would like to use the particles as tracer for FCS Microrheology. Thanks in advance.

Best Regards

Cihan

What are the merits of using Methyl Orange rather than other Dyes in photocatalytic degradation?

How to remove dyes like rhodamine and methyl violet from the textile effluents by electro coagulation and electro oxidation?

I am trying to create a workflow for flow cytometry experiment sample prep. I intend to use LIVE/DEAD™ Fixable Aqua Stain and an antibody cocktail for extracellular antigens.

I intend to use the viability dye first. The protocol I found does not seem to mind the protein content in buffers, as the cell washing is performed with PBS (5-10% fetal cell serum(I imagine they meant "calf")), and the cells are suspended in the same buffer before viability staining. I read that the presence of proteins in the buffers might contribute to a higher background, is that correct? How should I adjust the solution (for washing and resuspension) if that is correct?

Additionally, I intend to perform antibody titration for the antibodies I'm using. There are a few questions here too. We have no prior experience with these antibodies in the lab. Questions:

1. Is it important to do titration for every antibody in the cocktail?

2. Do I keep the same general sample prep with the antibody cocktail, only swapping the antibody cocktail with a single antibody that's in different concentrations? Do I also apply the viability dye as well?

3. The antibody cocktail I am to use includes dyes such as SB and BV etc, which require a special staining buffer or blocking buffer. Do I need these special staining buffers or blocking buffers for titration as well? Is it necessary since titration typically only uses one dye at a time (together with viability dye?)?

I know the information above might not be detailed enough but could you share your personal experience related to these scenarios? The protocol that I base my protocol on comes from this page

Thanks a lot for your input!

I'm attempting to stain cancer cells in suspension from a 12-well plate for my research project. Could anyone provide guidance on the most effective staining protocols and techniques for ensuring accurate and reliable results? Any insights or recommendations on suitable staining dyes, concentrations, fixation methods, and imaging procedures would be greatly appreciated. Thank you in advance for your assistance!

Hello, I have nylon fabric dyed with Acid dye. I need to prove for part of my project, is it possible or not possible, the reaction between dye molecules absorbed in nylon fabric with water molecules when the fabric is immersed in water. I hope someone helps me to find the best answer to this question.

We developed a rapid viability test for plant seeds in which yeasts metabolize organic compounds that leach from seeds (dependent on the physiological age of the seed) and thereby reduce our redox-indicator resazurin, accompanied by a color change (5). Unfortunately, seeds of some species acidify our test solution causing a pH-dependent colour change of the resazurin (abiotic reaction) which distorts our test results (6). We are looking to replace our redox indicator dye for these cases!

We already researched on several dyes from the tetrazolium-family that unfortunately also change their colour upon acidification, i.e., MTT (1), TTC (2), MTS (3) or the solubility is severely limited, e.g. MTT (3), XTT (4).

We are grateful for any advice!

1: Plumb et al. (1989) Effects of the pH Dependence of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide-Formazan Absorption on Chemosensitivity Determined by a Novel Tetrazolium-based Assay. Cancer Res 49: 4435–4440.

2: Lopez Del Egido et al. (2017) A Spectrophotometric Assay for Robust Viability Test of Seed Batches Using 2,3,5-Triphenyl Tetrazolium Chloride: Using Hordeum vulgare L. as a Model. Front. Plant Sci. 8: 747. doi: 10.3389/fpls.2017.00747

3: Riss et al. (2013) Cell Viability Assays. https://www.ncbi.nlm.nih.gov/books/NBK144065/

4: Goodwin et al. (1995) Microculture tetrazolium assays: a comparison between two new tetrazolium salts, XTT and MTS. J Immunol Methods 13: 95-103. doi: 10.1016/0022-1759(94)00277-4

5: Mohammed et al. (2019) Dead or Alive: Simple, Non-destructive, and Predictive Monitoring of Seedbanks. Trends in Plant Science 24: 783-784. DOI: 10.1016/j.tplants.2019.05.014

6: Wellmann et al. (2023) Maize Grain Germination Is Accompanied by Acidification of the Environment. Agronomy 13: 1819. doi.org/10.3390/agronomy13071819

In DSSC, TiO2 photoanode with Nb2O5 blocking layer (FTO/TiO2 interface layer) can prevent electron recombination due to its higher CB edge than TiO2. But, how will electrons be injected from TiO2 to Nb2O5 or how will electron transfer from lower CB to higher CB?

Dear all,

We have a Olympus FV1200 confocal microscope with 405, 473, 559, and 635 lasers.

I'm looking for a nuclear dye (fixed cells) in the far red range, does anyone have any recommendations for a good dye? The next closest fluorophore we routinely use is Alexa Fluor 594

Many thanks,

Sam

Colleagues, tell me which dye is optimal for staining RNA during gel electrophoresis? Which one do you use in your laboratory?

While doing photocatalytic dye degradation i observed some materials decreases the concentration of cationic dye but increase the concentration of anionic dye.

I am running SDS page gels using the mini-PROTEAN tetra vertical tank. Gels ran normally up until about 3/4 of the way down the gel. At this point, the dye changes from blue to yellow and became distort. I have inspected the module so this is not the problem. The electrophoresis buffer used has the correct pH. The loading dye was Laemmli Loading Buffer. All gels were run at 140V for 50-60 minutes.

I have an experiment involving acridine orange/ethidium bromide dual dye (AO/EB). How can I safely eliminate or inactivate these toxic chemicals afterward? and how to clean up the lab in case of any accidental spills or to remove any possible traces? Is using soap and water (or any detergents) enough?

I am currently in the process of doing a Western Blot with mESCs after an AHA Assay. I added 25uL of 2xLaemmli buffer and 25uL of AHA buffer (50mM Tris HCL) to my pellet.

I usually denature my proteins before loading on SDS Gel by boiling the sample with dye at 95C for 5 min. However, this time I forgot to take only half of my sample and store the other half for a repetition in the future. Instead, I added the dye and denatured the protein lysates by boiling. Can I still load half of my sample and save the rest in -80 or -20? Or should I leave it at RT? Or finally, should I just discard the rest of my samples?

I will appreciate some help!

I did FTIR of a reactive dye's powder. But I see that the transmittance of 3 peaks went opposite direction of normal peaks and eventually the transmittance percentage became more than 100%. What could be the cause of this?[ Please see that attached figure]

Hello ResearchGate Community,

I am searching for a water-soluble dye that can withstand a prolonged period (one month) at 85°C and 5000 psi. It's crucial that the dye remains stable and retains its color across both acidic and basic pH environments. The dye's ability to maintain consistent color under these extreme conditions is essential for my research.

I would appreciate any recommendations for dyes known for their stability and color consistency in such settings.

Thank you in advance for your insights!

Best,

Musa

I am trying to measure uptake of PLGA nanoparticles encapsulating DiI dye. I see the uptake just fine but my dish is covered with huge chunks of DiI-labelled debris. I wash 3 times with 2 mls PBS each wash but the chunks are everywhere and they move around which complicates downstream processing. Has anyone experienced this? How to get rid of it?

Hello all dear

I need a help

Thanks in advance

Need this for my research please also indicate which article it was from, thank you.

Hello!

I treat a polymer surface with plasma and I expect that some free radicals form on it, because it demonstrates high adsorption. How could I quantify the presence of free radicals on a polymer surface? Is there a simple assay for this purpose (a dye that changes color or fluorescence)?

We would like to mark nocturnal animals with something that we can see with an infrared camera, and ideally use in a way to be able to differentiate between animals. Does anyone have any experience or suggestions of something we can use?

I'm trying to identify a marker/dye that can be assessed (via histology) from post-mortem eyes which have undergone subretinal and suprachoroidal injections. Any advice on successful agents would be useful.

The binding of the sugar-dye compound with a lectin should facilitate a shift in the emitted light that enables monitoring of lectin activity. Or perhaps there are other ways to monitor lectin activity apart from a heamagglutination assay?

I loaded the 1.5% agarose gel as standard. I loaded 6uL from PCR and 1uL loading dye. Why is the solution not sinking to the bottom of the wells?

If I add a dye to a liquid, and this liquid is subjected to a high temperature may reach 110 oC does this temperature will affect the dye colour or not

I am trying to load calcium Green Tm AM inside the neurons. As a first step, I would like to ensure the dye permeates the cell membrane. I am using a 5x and 10x objective to quantify the background fluorescence, then, I inject 200ul of a solution containing the dye and get some pictures post-injection. There is a difference from the background image, but I don’t have a cellular resolution. Would anyone have any suggestions on how I could approach this?  I am not sure, for example, whether I can image the dye (it could also be fluo-4, I have a number of those, and I am trying to establish a protocol to check their levels of cell permeability) post PFA fixation using a confocal microscope.

Thank you very much for your time!

For organic liquid dye preparation to be used in cosmetic formulations

Anyone knows exact dye name with code and other information. Reactive orange with 418 nm wavelength.?

What is the rationale for an adsorbent to adhere to both the Langmuir and Freundlich isotherms in the elimination of methylene blue dye, while remaining unaffected by changes in temperature?

FACS

I want to know if there is any low cost method to identify an exopolysaccharide produced by bacteria is cellulose or other polymer.

by relative method

To decrease the wastage of Reactive Dyes, its Hydrolysis must be reduced or removed

I only used 2 wells of 12-well gel for my first electrophoresis. Can I reuse the remaining wells at the end of this electrophoresis? Could the gelred dye in the gel have been affected by the previous electrophoresis? In my view, the dye molecule cannot move without binding with DNA, so I can use the free well one more time. But someone told me I should turn around the gel if I wanna use it one more time to avoid the influence on gelred dye after the first electrophoresis.

I am conducting an experiment where I am putting cultured cancer cells (B-ALL line) in whole blood and running them through a microfluidic device. I stain the cells with DAPI and DiI (cell membrane dye) and then add them to whole blood. The issue that I am facing is that clumps develop in my microfluidic device when I run the sample. I have noticed that the amount of clumps and the time it takes for them to develop are related to the concentration of cancer cells I add in the blood. (The more cells I add, the bigger the clumps and the quicker they develop.) Does anyone have experience with spiking cancer cells in whole blood and recommend any changes? Should I fix the cancer cells before spiking?

Hello all dear

Can you introduce me some literature about this?

Thanks in advance

I wish to study dye degradation using these nanoparticles but I am unable to find adequate research papers on biosynthesis of iron-selenide nanoparticles.

Looking for technical guide on better dye to stain protein in starch samples for study unders confocal laser scanning microscopy.

Hi everyone! I am new in lab and I have been having problems with Western Blot, I use a Chemidoc and when I reveal I see nothing, after reincubate, or incubating with a new antibody, the signal is lost, or, is very very low, when I dye with red Ponceau, I see a lot of protein because I put 40 ug per lane, I don't have idea about what happened, someone could help me, I will be eternally grateful

HK green eyes are most suitable for the same but they're not available anywhere that's why I'm asking the question.

I have been having issues with compensation whenever I use 3 brilliant violet (BV) dyes together for flow cytometry. I heard BV buffers are the game changers but they are quite expensive. So I am looking for a substitute.

I performed photocatalytic degradation experiment under direct sunlight. The original dye solution was showing lesser absorbance value than the solution kept under direct sunlight in presence of photocatalyst. How can I improve it?

I have prepared an azo dye with orange colour in polar solvents. As is seen in the absorption spectra, absorption is in the UV region and up to around 400 nm in the visible. However, an orange colour solution typically shows absorbance in the 500-600 nm range. This dye also shows a green specular reflectance in a dry solid form. Could anyone please explain the possible reasons for these two phenomena?

I tried staining beads with zombie aqua dye. but the beads did not stain at all. I use 1:400 dilution for my cells. I used the same dilution for the beads as well. do the beads do not stain at all with zombie aqua or do I need to increase the concentration?

Hi,

I am currently using pHrodo Green AM as a marker for my target cells, which was used in a phagocytosis assay with my M1-like THP-1 cells and I have some questions regarding this specific dye:

(1) pHrodo Green AM was said to be a dye that is able to emit fluorescence on low pH conditions. However, when I observed the fluorescence using flow cytometry (FITC channel), I was able to see some strong fluorescence from the labeled target cells directly. Should this be happening? Considering that pHrodo dyes are supposed to only react with acidic conditions.

(2) I also perform co-culture between the target cells and the M1-like THP-1 (labeled with another dye). However, I saw that after co-culture, there seems to be a decrease in pHrodo Green fluorescence instead of an increase. Has anyone else observed this pattern?

Thank you.

??

Kindly give any reference article

I am interested in showing innervation in the anterior chamber of the eye, in live mice. Is there any dye I could use?

What is the best dye to use for Automated Liquid Handling System dispensing verification? Currently i am using Orange G. but if you have a good data with other dyes, please share it with me. Thank you.

Removal of dye

what can be the working conc of the dye in a 96-well plate experiment?

Concerning the adsorbent dose for example iron nanoparticle to remove dye. Sometime it is written as

1- "the run was conducted by adding different dose of iron from 10 to 150 mg to 50 ml working solution of 30 mg/l of dye".

Other research paper mentioned

2- "the run was conducted by adding different dose of iron from 10 to 150 mg/l to 50 ml working solution of 30 mg/l of dye".

In case #1 it is clear that I have to add 10 to 150 mg of adsorbent to 50 ml. But I'm a bit confused in case #2. Should I have to prepare adsorbent solution with that concentration?? Or 10 to 150 mg of adsorbent will be calculated in 50 ml of adsorbate, i.e. in case of 10 mg of adsorbent dose, I have to add 0.5 mg of adsorbent to 50 ml working solution