Cell Culture Techniques

Cells in culture or in vitro are a useful model for studying the activity of cells in the whole organism or in vivo. Ten years ago or so cell... | Contact experts in Cell Culture Techniques to get answers

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Questions related to Cell Culture Techniques

How do i improve my growth rate of EBV- B cell LCLs?

Hello al l I have an Epstein-Barr Virus- (EBV-) Immortalized Lymphoblastoid Cell Lines (LCLs) which grow very slow. the media I using is 10%RMPI or DCCM1 how can I improve my growth rate? thanks...

09 September 2021 3,777 1 View

Help regarding XCMS?

I am a beginner to learn about XCMS. For my plant sample, I have already done LC-MS. I want to identify a few of the peaks. after searching some literature I came to know about XCMS. I read some...

08 September 2021 1,771 4 View

Should I be concerned when my BJAB cells adhere?

I'd like to work with BJAB cells. As I understand, they should be growing in suspension. However, I observe some adherent cells in my cultures. I already thawed several aliquotes from our frozen...

07 September 2021 8,833 0 View

Understanding replicates and best statistical test?

Hello, I am trying to analyze some pilot data with the following setup: I basically have three different types of drugs which were administered to my cells at 3 different doses (9 conditions, if...

19 August 2021 2,588 7 View

Despite being a suspension cell line, why does the NFS-60 cell line cling to the culture flask surface?

I'm now dealing with the NFS-60 cell line, which is a suspension cell line, however, it continues to grow and shows that it is attached to the culture flask surface despite the fact that I changed...

17 August 2021 3,919 3 View

Can you recommend a good and reliable (inexpensive) pipetting robot suitable for a small lab?

Dear colleagues, I am looking for a pipetting robot, primarily for preparing 384 well plates. There are plenty devices on the market but I found the Eppendorf epMotion 5070 as well the less...

15 August 2021 7,845 0 View

Precipitation Issue with Rotenone?

Has anybody had any issues with Rotenone precipitating out in cell culture medium. I see brown precipitates forming in cell culture media after 1-2days in culture at doses above 30uM. If so, what...

29 July 2021 3,976 0 View

Why my glycolysis seahorse data looks so strange?

Hi, guys, recently I am performing the glycolysis seahorse assay in endothelial cells. I followed the protocol as suggested, but the ECAR didn't increase after adding Oligomycin. Do you think the...

19 July 2021 3,181 4 View

Reaction conditions to functionalize hydroxyl group with methacrylate?

Hi everyone! I am trying to functionalize the hydroxyl group of poly(ethylene glycol) with methacrylate by using methacrylic anhydride. I was wondering if anyone has any recommendation regarding...

15 July 2021 6,283 3 View

Bad Ponceau S staining with PVDF membrane?

After taken out from transferring, I wash the PVDF membrane with 1X TBST for 5 minutes before staining it with used or new Ponceau S. I then rinse it with dH2O to get a clear background. However,...

09 July 2021 574 4 View

Why is my nucleus being fuzzy under confocal with hoechst staining?

I am wondering why my hepg2 cells nucleus looks fuzzy and unclear under the confocal? Is this mycoplasma contamination? I used Hoechst staining, 1:200 dilute with 3%BSA.

26 June 2021 7,434 3 View

What is the best protocol for paraffin-embedding mouse brains?

We have mouse brains that have been sitting in PFA for a couple of months, and we now wish to paraffin-embed these PFA-fixed mouse brains for immunofluorescence microscopy. We have an automatic...

24 June 2021 7,969 5 View

Why Vero cell are detached from 24 well plate after fixing ?

I am doing plaque assay for CHIK-V virus. After incubation plate was wash fix with 3.7% formaldehyde sol. After fixing cell was detected. Before fixing monolayer was fine.

23 June 2021 9,050 6 View

How can I measure protein concentration without a plate reader device?

I have a Bio-Rad ChemiDoc XR+ and a Pierce BCA Protein Assay Kit. I am wondering if I can do the Pierce BCA Assay without a plate reader? Or if I can use the ChemiDoc XR+ for the assay as a...

15 June 2021 2,115 5 View

LIMS software recommendations?

I am interested in a software/solution which will allow employees across the company to submit request for various analyses in the labs, allow the lab managers to track the samples and work load...

11 June 2021 10,005 4 View

MRNA isolation from acutely isolated microglia, tips?

Hi everyone. I am going to isolate microglia from 2 months old mice using the Percoll gradient (30, 37 and 70%), for subsequent FACS sorting and mRNA isolation for RT-qPCR on inflammatory...

02 June 2021 5,414 6 View

Why did my mESc are floating and dead?

Hi, I recently cultured the AES0186-BRC-6 Feeder free mESc. When I cultured the cells, they were good until the second passage, and at the third passage, the cells died. I used the following media...

31 May 2021 8,996 1 View

How to use amphotericin B in contaminated cell cultures?

I am using ovarian cancer cell lines and they were heavily infected by fungal spores? What is a good conc. Of amphotericin B to use? And how to apply?

30 May 2021 3,130 4 View

We want to make l- glutathione face wash pl give any suggestion if anybody else?

composition 1. glutathione 2% 2. kojic acid 2% 3. ALFA arbutin 2% 4. ascorbic acid 10% 5. vitamin e oil 1%

24 May 2021 4,580 4 View

Application of UMAP in flow cytometry?

I want to know how I use the UMAP plots for my flow cytometry data.

22 May 2021 1,952 2 View

What should be the concentration of the sterile saline for the inoculum preparation for Disk-Diffusion Assay and Broth dilution method?

I need to know what concentration of the saline should be for the preparation of E.coli and S.aureus for disk diffusion assay and broth dilution method. Does the concentration even matter, if it...

22 May 2021 5,442 2 View

How can the forward and reverse sequence of an isolate be combined for a BLAST analysis to be performed?

Recently I sent amplicons generated from isolates of bacteria for sequencing. The sequences received (forward and reverse) are not combined and blasting of these sequences need to be performed....

19 May 2021 3,019 4 View

How can I quantify the number of two color LC3 particles with imageJ?

Hello, I am using RFP-GFP-LC3B for autophagy flux detection. After merge RGP-LC3 and GFP-LC3 result to a merged pic with yellow and red dots of LC3. I wonder how to seperately count red and...

18 May 2021 5,525 3 View

Questions about Raw 264.7 cell culture..?

Dear everyone, I am relatively new to cell culture, and I need some help. At the moment, I am culturing RAW 264.7 cell line. I have some question regarding this. I am not quite sure about the...

18 May 2021 3,433 14 View

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