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Questions tagged [cell-culture]

Cell culture is the process of growing and maintaining living cells in vitro.

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How do I grow 5 liters of purple non-sulfur bacteria?

I am running experiments for a project that requires large amounts of purple non-sulfur, Cyanobacteria and nitrogen fixing bacteria. I plan on growing all the species in separate tanks. The species' I ...
Aakarsh Tathachar's user avatar
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How can I be more accurate in my cell counting?

For some reason, when I use my hemocytometer, my counts and calculations always come out really small. My equation is: (cell count / # of boxes) * dilution factor * 10,000 = cells/ml. (I usually only ...
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Any tips to reduce cell death (from thawed fibroblast cell cultures)?

I have been doing cell culture for about a year now. When I thawed my fibroblast cell line, everything went to plan. When I checked them a day later, they were all dead (<3-5% confluent). I'm not ...
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What is the largest culturable bacterium?

E. coli is easy to grow in the lab. It has a volume around $0.6 \mu m^3$. Source T. namibiensis can be much larger, with a volume on the order of ${10^6}\mu m^3$. Source However, T. namibiensis is not ...
julianstanley's user avatar
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Cell viability assay: Problems with MTT assay in the solubilization step

I am testing two compounds against 3 cell lines to determine cell viability. Two lines grow in Eagle media, one in DMEM (+10 Percent bovine serum) which contain phenyl red indicator. I prepared 3 ...
raptorlane's user avatar
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Why does my cell subculture have these black dots like appearance?

I tried to subculture cells(hek293 cells) for the first time for a Cell Biology experiment, and here are the steps I followed:- Cell subculture (*the media is a mix of 10:1:0.1 of DMEM:FBS:Pen/Strep) ...
Khushi Parashar's user avatar
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For Cytotox studies of anticancer compounds: What cell line can I use to check if my compounds will also damage healthy cells?

I am about to perform cytotox studies on cells with my synthesized compounds. I want to test my structures against MCF7 cells (breast cancer) as well as a melanoma cell line. MCF7 should be vunerable ...
raptorlane's user avatar
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Plant tissue cultures without Phytohormones?

Is it possible to do tissue cultures without using phytohormones like BAP or NAA. So to just use for instance MS-Media. Online, I just found that every plant is possible to culture with the right ...
Jonah's user avatar
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Is this contamination, debris?

I have been culturing SVG cell lines. This line was obtained as a live culture from another institute and was delivered by mail. After subculturing , I saw some particles, and I do not know what they ...
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How much effort is it to establish a cytotox assay for cancer cell lines against a small number of possible compounds?

I am currently testing a series (5-10) of small molecule compounds against an enzyme that are intended as inhibitors. This enzyme is meaningful for cell proliferation. Until now, nothing was active ...
raptorlane's user avatar
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How to keep primary human cancer cells alive in vitro and derive a cell line?

I am studying a human cohort of patients treated with neoadjuvant immunotherapy before surgery, of whom we have pre- and post-treatment biopsies as well as the resection. In one patient's resection, ...
immunoLogical's user avatar
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Why are there error bars for zero concentration in some cell viability graphs, if they are assumed to be 100%?

Take these for example: As far as I know, the formula for cell viability is % Viability = (Treatment OD - Blank OD) / (Control OD - Blank OD) where each OD is the average of three wells (technical ...
h088bmIuXaskpzJEe3ld's user avatar
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Bacterial cells disappearing from culture with time

While growing my bacterial culture in liquid media for protein expression, I have been observing that the bacterial cells (E. coli) grow well for some till OD600 reaches ~0.4-0.6, then suddenly ...
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Cell culture disasosiation

I am doing to passaging cancer cell ( Dld-1 Cell line). Trypsination was conducted with 0.25 trypsin-edta 1 ml to t-flask 25 in 2 min. Then I add 6 ml complete media (have 10% FBS for inactivated ...
MUHAMMAD SHOFI's user avatar
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Can a non-pure culture be used for reliable rapid phenotypic diagnostic tests?

Can a non-pure culture be used for reliable rapid phenotypic diagnostic tests (e.g. catalase, oxidase & gram stain)? "Slide Test Method Obtain a pure culture of the organism to be tested. ...
Freezing Soul's user avatar
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Induction of IFN-beta in HEK293T

I'm trying to increase expression of a protein we're attempting to study, UBL7, supposedly unregulated by Type I Interferon and particularly IFN-beta. I've tried treating HEK293T cells (~60% ...
Tom Murphy's user avatar
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Where can I find a video of human cells reacting to cold water vs warm?

I have the following lists: ...
Andrew Arrow's user avatar
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What are common culture temperatures?

In literature, authors sometimes refer to 37°C as a common culturing temperature. As I understand it, those are temperatures that are conventionally used for incubators in laboratories. For mesophilic ...
kate allerton's user avatar
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What kind of bubble this?

This is the MARC-145 I cultivated. Recently, many bubbles appeared in the cells. I don’t know why.
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Cell culture contamination

I've been culturing my cells over 2 weeks and just yesterday these things popped up in my flask , I'm not sure if they're contamination or not.uesterday they looked white and today I found one of them ...
Neda's user avatar
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Need help identifying this microbe

Sorry for the bad image, we don't have a camera hooked up to our culture room microscope yet. But Has anyone seen these kind of (what I'm assuming are) contaminating bacteria before? We see these from ...
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Is somatic fusion/hybridization off-limits to DIY-ers? ELSE: how can an infertile plant's characteristics be salvaged to a fertile one?

I was amazed to discover plant tissue culture could be performed relatively easily at home. I have been wondering, what's the likelihood of a DIY-er being able to setup and learn a crude and basic lab ...
geeheeb's user avatar
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Why is my spirulina biomass getting destroyed in my new photobioreactor?

I have a 500L flat plate photobioreactor in my backyard. Whenever I add my culture to the PBR, after 3-4 days of growth, the culture becomes transparent and my biomass is wasted. I add the stock at a ...
Milo's user avatar
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Does decellularization with sodium lauryl sulfate break down enzymes?

I am currently looking into decellularization with sodium lauryl sulfate since it is effective and cheap. My goal is to break down the cells of a plant leaf and release the chloroplasts to be viewed ...
Aakarsh Tathachar's user avatar
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Endospores contact with surfaces

I am currently studying endospores-forming bacteria. To be specific B.subtilis. I noticed that the formed spores have strong contact "grip" with the surface it is forming on. For example, ...
Anwar Elhadad's user avatar
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Single-cell subcloning of BV-2 cells?

I am trying to knock-out genes in BV-2 cell line. However, the majority of protocols require cell subcloning and expanding. I tried to subcone these cells and grow clones in DMEM+10% heat-inactivated ...
Monika1223's user avatar
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MTT assay data pre-processing

I was handed MTT assay data, consisting of drug doses and raw OD scores. The drug treatments were performed three separate times, each followed by MTT assay. I applied some non-linear mixed-level dose-...
Bryan's user avatar
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What is the solubility of rotenone in ethanol?

I’m trying to kill SH-SY5Y cells with rotenone, but the rotenone does not kill. I saw an article where rotenone is soluble at 5mM in ethanol. I dissolved 1.7 mg in 1 ml of ethanol to obtain a 1ml of ...
user73512's user avatar
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204 views

Is there a way to relate the OD to cells/mL without doing a standard growth curve?

I am helping out with an experiment with multiple bacterial cultures (>20 species). I need to a very specific concentration (cells/mL or CFUs*/mL) for this purpose. Unfortunately it appears that ...
Dunois's user avatar
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Does centrifuging the bacteria culture removes bacteria leftovers?

Usually, we centrifuge bacteria culture to either replace the media, control its density (OD), ..etc. I know that when centrifuging the cells are pushed to the bottom while the media (solution) ...
Anwer Ak's user avatar
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What is the difference between BSA fraction V and the initial fraction?

I am preparing a supplemented culture medium and I need to use BSA. There is fraction V of BSA that we normally use for immunocytochemistry and there is another type named the initial fraction that is ...
Helen Etemad's user avatar
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Fixing the cell with 4%PFA and staining with PI to observe cell cycle?

I am wondering whether we can replace 70% ethanol to fix and perm the cells with 4% PFA to observe the cell cycle. Do I understand correctly that 4% PFA can permeate the cells, thus the PI will ...
tassaneel's user avatar
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Virus culture in artificial/synthetic medium

I am looking for a publication/paper in a well a circulated magazine/journal/government study on growing virus culture in synthetic or artificial medium. I have found this link on SARS Cov 2 being ...
Cryptex's user avatar
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What is the meaning of harboring cryptic in below sentence?

Tissue culture became more widely used at this time because of the introduction of antibiotics, which facilitated long-term cell line propagation, although many people were already warning against ...
Arash Salehi's user avatar
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Appropriate cell lines to study depression

Short introduction I'm studying depression from a biochemical point of view. My interest lies in the study of protein biomarkers and I was wondering which cell lines may be appropriate for this ...
Sam's user avatar
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How do I cover Cellstrainers?

I‘m doing some experiments for my bachelors thesis and I‘m using some Cell strainers by Roth for it (https://www.carlroth.com/com/en/accessories/cell-strainer-easystrainer™/p/cly9.1). My problem is ...
David Harvest's user avatar
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1 answer
181 views

Sabouraud dextrose agar breaking?

I recently started growing some Geotrichum Candidum (GC) on Sabouraud dextrose agar (SDA) beds. The SDA was poured into sterilized petri dishes. After setting the plates in a cooler (20 C) for a day, ...
neydroydrec's user avatar
1 vote
1 answer
258 views

How to create long term stock for Bacillus subtilis?

I am trying to create a long-term stock for Bacillus subtilis (Bacterial glycerol stocks). I have looked online for guides and step-by-step tutorials and I found that they don't say where the sample ...
Anwar Elhadad's user avatar
6 votes
1 answer
58 views

dilutions in bacterial culture

In overnight bacterial culture, why do we put bacteria in a small flask to culture before moving it to a larger flask? Why not just put the sample in a large flask to begin with? This is in reference ...
Val's user avatar
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Decreasing signals in assay measurements

I'm working with a calcium assay to study the effects of different virulence factors. The assay works, but from day to day the signals of cell lysis go down. Unfortunately, I haven't found an ...
Mourinho_1's user avatar
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CML (Chronic Myeloid Leukemia) and blast cell percentage

I'm trying to understand the oncogenesis of CML. I have a question about CFC cells and blast cells. Are those the same? I know that in the chronic phase of CML there is a blast percentage of 1-10% of ...
Mario Pérez's user avatar
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QA/monitoring in recombinant vaccine manufacturing

On a16z[0] podcast on vaccines[1], an interesting tidbit came up: vaccine manufacturing in cell cultures is expected to hit a major challenge in terms of quality control due to unpredictable behaviour ...
Grzegorz Kossakowski's user avatar
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2 answers
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Why does vacuum sealing of foods prevent spoilage from anaerobic bacteria?

I have noted that many purchased food items in containers only require refrigeration after they have been opened, thus eliminating the vacuum seal. Air contains bacteria and fungus spores that enter ...
0tyranny0poverty's user avatar
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types of cells in a pharyngeal specimen

I would like to ask what type of cells should someone expect in a pharyngeal smear, in a person, who is infected with a viral infection. Thank you a lot!
marilu's user avatar
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Would BMP7 introduced to a Petri dish with cartilage cause chondrocytes to replicate?

Chondrocytes are the cells that make up cartilage. As I understand it, the best growth factor that signals replication of chondrocytes is Bone morphogenetic protein 7. If supplied with a sufficient ...
Delaney Fitzpatrick's user avatar
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Public fermentation/cell cultivation database?

I am wondering if there are any large collections of fermentation or cell cultivation data publicly available. I am interested in all cell types used within industrial biotech and biopharma, ...
Toke Faurby's user avatar
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1 answer
159 views

how to culture mc38 cells

I work at a start up company in a vivarium. My boss would like me to start working with MC38 Cells. I bought all of the equipment (recommended by a colleague) and should be ready for the cells. Could ...
msmith's user avatar
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Cell image database for citotoxicity test

Is there any cell image database for citotoxicity tests? I know that there are several image databases available (like this, this and this one), but none of them have the images for citotoxicity tests....
Gabs's user avatar
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1 answer
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Image Dataset for Cardiac cells for Mus musculus?

I'm working on developing a transfer learning model to detect cells in a culture using microscopic images. for that i require data set of images having microscopic images of various cell types. Is ...
himanshu saraswat's user avatar
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Why do coliform bacteria smell so bad?

I have done multiple cultures of E. coli bacteria and related species over the course of my education. Almost every time, they had this revolting smell. However, other organisms used for genetics ...
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