Can anyone suggest for Propidium Iodide (PI) and DAPI dual staining protocol for cell culture?

More Elham Amini. E.A's questions See All

Promotor observation in region annotation of RNA RIP-seq?

Hello all, I extracted RNA from my samples and performed RIP-seq. After annotating the genomic regions using R, I obtained promoters, exons, introns, and UTRs. Given that my samples consist of...

18 July 2024 1,579 2 View

Why do I get no melt curve peak and no Ct for some of my patients but not others, in a SYBR green qPCR?

I am doing a RT-qPCR for gene expression analysis of cancer patients, and there are four different groups in the results. The first group that shows over expression of my target gene (both two...

31 May 2024 1,088 0 View

Can anyone interpret why the absorbance in alpha amylase assay get increased with increased standard (inhibitor) concentration?

Can anyone interpret why the absorbance (DNS assay method) in alpha amylase assay get increased with increased standard (inhibitor) concentration? it should be decreased with increased standard or...

26 May 2024 4,753 2 View

Anyone with phosphate flotation experience?

I attempted a phosphate (apatite) flotation, but the slurry was too thick for the impeller to work properly. I reduced the solid content to 30% for better mixing, but the conditioning still seems...

14 May 2024 4,505 4 View

Why my amino acids do not contribute in amidation reaction?

Hello everyone, recently I am trying to synthesize a dipeptide with a Coumarin derivative of Tryptophan, and the problem is it doesn't contribute in reaction at all. Tryptophan itself in the same...

15 April 2024 6,778 3 View

FinFET-10nm Technology File?

Dear Authors and Researchers, I'm looking for predictive technology models (PTM) for FinFET including 10nm. Unfortunately, the following webpages do not work...

14 April 2024 4,390 1 View

Is there any technical way to test stability analysis of wave function in QUANTUM ESPRESSO?

I did a branch of the DFT calculation to find the stable cell parameter (a) . I didn't get the smooth curve for energy(E) v.s a. There is some point out of concave curve that I had to repeat...

21 March 2024 2,037 2 View

How to increase concentration and quality of RNA after extraction?

Hello All, I'm doing RNA extraction from cells. I tried both ways by using TRIZOL (with chloroform, and isopropanol) and RNeasy mini kit. When I use TRIZOL, the RNA concentration is high, however...

11 March 2024 9,171 8 View

Why a new peak has appeared in the spectrum after dye adsorption?

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...

10 March 2024 4,167 2 View

How to interpret my samples' results from D1000 ScreenTape Assay?

Hello All I prepared my library to run for sequencing. I would like to check the quality of my samples through D1000 ScreenTape Assay before add to sequencer. Can anyone help me to interpret my...

10 March 2024 8,764 0 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,105 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 640 2 View

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Can I multiplex a mouse monoclonal and rat primary for IHC (mouse brain tissue)?

Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...

06 August 2024 9,708 1 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

How to increase citation in Research Gate?

How to enhanced h-index in Research Gate?

04 August 2024 3,367 4 View

Subhash C. Juneja

May be you should do PI staining on live cells, then fix cells for 15 min in 4%PFA and treat cells with DAPI for 15 minutes and wash with PBS 2 times, hope that works

Elham Amini. E.A

Thanks for your reply.

Actually, I don't want to fix my cells. Is there any way to use PI and DAPI without fixing the cells?

Sergei Rudchenko

Both fluorochromes do not penetrate through membrane of live cells. These fluorochromes are used for staining dead cells for live/dead discrimination.