Log In Sign up
The largest database of trusted experimental protocols

Trypan blue solution

Manufactured by Fujifilm
Visit Supplier
Sourced in Japan, United States

Trypan blue solution is a laboratory reagent commonly used in cell counting and viability assays. It is a dye that selectively stains dead or damaged cells, allowing researchers to distinguish viable cells from non-viable ones under a microscope.

Automatically generated - may contain errors

Lab products found in correlation

Countess 2, by Thermo Fisher Scientific (3 mentions) Trypsin edta, by Fujifilm (2 mentions) Tc20 automated cell counter, by Bio-Rad (2 mentions) Trypan blue, by Bio-Rad (2 mentions) Trypsin edta solution, by Merck Group (2 mentions) Bz 8100, by Keyence (1 mentions) Fitc labeled anti cd45 antibody, by BD (1 mentions) Facscalibur, by BD (1 mentions) Staurosporine, by Abcam (1 mentions) Casp3, by Cell Signaling Technology (1 mentions) Chondroitin sulfate, by Seikagaku (1 mentions) Z vad fmk, by Peptide Institute (1 mentions) Z vad fmk, by Thermo Fisher Scientific (1 mentions) 12 well plate, by Thermo Fisher Scientific (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Chloroform, by Fujifilm (1 mentions) Sodium deoxycholate, by Fujifilm (1 mentions) Isopropyl alcohol, by Fujifilm (1 mentions) Sodium dodecyl sulfate (sds), by Fujifilm (1 mentions) Hygromycin b gold, by InvivoGen (1 mentions)

Spelling variants of this product

Trypan blue (21 citations)

19 protocols using trypan blue solution

1

Cell Viability and Transfection Efficiency Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell viability was examined using the trypan blue exclusion assay as previously described [28 (link)] with slight modifications. Cells were re-suspended in culture medium (100 μl) in a well, and a small portion (3 μl) was mixed with an equal volume of 0.4% trypan blue solution (Fujifilm Wako Pure Chemical, Osaka, Japan) in a 1.5-ml microcentrifuge tube. The mixture (6 μl) was then transferred to a hemocytometer (Watson, Tokyo, Japan) and the number of viable (unstained) and dead (stained) cells was counted under a microscope (IX70, Olympus, Tokyo, Japan). Three replications (three wells) were made for each incubation period, and the wells from which cells were taken were sacrificed after each count. Cell viability was calculated by the ratio of viable cells to total cells.
Enhanced green fluorescent protein (EGFP) fluorescence was observed under a fluorescent microscope equipped with a GFP filter (BZ-8100, Keyence, Osaka, Japan). The EGFP-positive and -negative cells observed within randomly selected areas of 200 μm x 200 μm squares under a fluorescent microscope were photographed and counted to estimate transfection efficiency (EGFP-positive cells among viable cells). The number of cells in three square areas per well was counted and three replications (for three wells) were made for each measurement.
Watanabe K., Yoshiyama M., Akiduki G., Yokoi K., Hoshida H., Kayukawa T., Kimura K, & Hatakeyama M. (2021). A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis. PLoS ONE, 16(9), e0257770.
+ Open protocol
+ Expand
2

Quality Control of ADMPCs Transplant

Check if the same lab product or an alternative is used in the 5 most similar protocols
ADMPCs were examined for quality at the time of cell freezing, cell thawing, and just before transplantation. Quality control tests included cell number, cell viability, cell purity, and infection tests. Cell number and viability were determined by staining with 0.4% (w/v) Trypan blue solution (Wako Pure Chemical Industries). Cell purity was analyzed by flow cytometry using FITC-labeled anti-CD45 antibody (BD Biosciences), FITC-labeled anti-CD105 antibody (Ancell Corporation, Bayport, MN, USA), FITC-labeled anti-CD166 antibody (Ancell Corporation), and FITC-labeled isotype control mouse IgG1 antibody (BD Biosciences). Data were collected with a FACSCalibur (BD Biosciences). The culture supernatant was collected by aspiration and used for infection tests including a sterility test, mycoplasma negative test, and endotoxin test.
Takedachi M., Sawada K., Sakura K., Morimoto C., Hirai A., Iwayama T., Shimomura J., Kawasaki K., Fujihara C., Kashiwagi Y., Miyake A., Yamada T., Okura H., Matsuyama A., Saito M., Kitamura M, & Murakami S. (2022). Periodontal tissue regeneration by transplantation of autologous adipose tissue-derived multi-lineage progenitor cells. Scientific Reports, 12, 8126.
+ Open protocol
+ Expand
3

Apoptosis Induction in HEK293 Cells by SCH79797

Check if the same lab product or an alternative is used in the 5 most similar protocols
HEK293 cells were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin (10 U/mL), and streptomycin (100 μg/mL), and treated with 0.3 or 1 μmol/L of SCH79797 or 0.1% of DMSO (v/v) as vehicle for 3, 6, or 24 hours. After this period, cells were lysed with RIPA lysis buffer, and the ratio of cleaved Casp3 (caspase‐3; 1:1000; Cell Signaling) to Casp3 (1:1000; Cell Signaling) was evaluated by western blot analysis. As a positive control, we administrated staurosporine 1 μmol/L (Abcam) to cells to induce apoptosis and collected cells 4 hours later. In a cell‐viability assay, HEK293 cells were cultured in DMEM supplemented with 10% fetal bovine serum, penicillin (10 U/mL), and streptomycin (100 μg/mL), and treated with 0.1, 0.3, or 1 μmol/L of SCH79797 or 0.1% of DMSO (v/v) as vehicle for 3 or 24 hours. After this period, cells were detached from the culture dish using Trypsin‐EDTA (Wako), and then trypan blue solution (Wako) was added. The number of whole cells and stained cells were counted with a TC20 automated cell counter (Bio‐Rad), and the survival rate was calculated.
Yokono Y., Hanada K., Narita M., Tatara Y., Kawamura Y., Miura N., Kitayama K., Nakata M., Nozaka M., Kato T., Kudo N., Tsushima M., Toyama Y., Itoh K, & Tomita H. (2020). Blockade of PAR‐1 Signaling Attenuates Cardiac Hypertrophy and Fibrosis in Renin‐Overexpressing Hypertensive Mice. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 9(12), e015616.
+ Open protocol
+ Expand
4

Preparation and Characterization of Calcium-based Biomaterials

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Scallop shell powders were heated at 1450 °C for 4 h, then ground using a dry super grinder (Nano Jetmizer NJ-300-D, Aishin Nano Technologies Co. Ltd., Saitama, Japan), followed by cooling in a vacuum chamber. This provided BiSCaO with dry powder diameters of 3–9 (average 6 μm) and was purchased from Plus Lab Corp., Kanagawa, Japan. According to the manufacturer, the content of CaO in this BiSCaO preparation was 99.6%. BiSCa(OH)2 was obtained from Scallow, Kohkin Inst. Co. Ltd., Tochigi, Japan, had a dry powder diameter of 10–100 μm (average 46 μm), and the CaO and Ca(OH)2 concentrations were <5% and >90%, respectively. Sodium polyphosphate (Na-polyPO4; PP), sodium triphosphate (Na-triPO4; TP), 0.4 w/v% trypan blue solution, and 1 N HCl were purchased from FUJI FILM Wako Pure Chemical Corp., Osaka, Japan. Chondroitin sulfate and heparin were purchased from SEIKAGAKU Corp., Tokyo, Japan. Non-anticoagulant heparin carrying polystyrene (NAC-HCPS) [18 (link),19 (link)] and low-molecular-weight heparin/protamine nanoparticles (LMWH/P NPs) [20 (link),21 (link)] were prepared as previously described.
Sato Y., Ohata H., Inoue A., Ishihara M., Nakamura S., Fukuda K., Takayama T., Murakami K., Hiruma S, & Yokoe H. (2019). Application of Colloidal Dispersions of Bioshell Calcium Oxide (BiSCaO) for Disinfection. Polymers, 11(12), 1991.
+ Open protocol
+ Expand
5

OVCAR-3 Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
OVCAR-3 cells (1 × 104 cells) were cultured in 6-well plates as described in Cell line and cell culture. Cell counts were performed 24 hours, 48 hours and 72 hours after OVACAR-3 cells were transfected with RECQL1-siRNA or control siRNA (as control). To distinguish live and dead cells, cells were stained with 0.3% trypan blue solution (Wako Pure Chemical Industries, Osaka, Japan). Cells were counted in a hemocytometer. The cell count was performed in triplicate, and the total cell count and the number of dead cells were represented as averages.
Matsushita Y., Yokoyama Y., Yoshida H., Osawa Y., Mizunuma M., Shigeto T., Futagami M., Imaizumi T, & Mizunuma H. (2014). The level of RECQL1 expression is a prognostic factor for epithelial ovarian cancer. Journal of Ovarian Research, 7, 107.
+ Open protocol
+ Expand
6

Apoptosis Inhibition in ALDH1high Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
In Fig 3J and 3K, ALDH1high cells isolated after PKCλ KD for 48 h with targeted siRNA were plated in 12-well plates (Thermo) at a density of 2 x 104 cells/well and incubated for 24 h. apoptosis inhibitor (using the pan-caspase inhibitor z-VAD-FMK) was purchased from PEPTIDE Institute Inc. and dissolved in 100% DMSO, making a 200 mM stock solution. In Fig 4F, ALDH1high control cells or MDA-MB 157 PKCλ KO cells were plated in 12-well plates (Thermo) at a density of 2 x 104 cells/well and incubated for 48 h with 100 μM z-VAD-FMK or 0.5% DMSO as a control. After staining with 0.4 w/v% Trypan blue solution (Wako), the cells were counted manually.
Nozaki Y., Motomura H., Tamori S., Kimura Y., Onaga C., Kanai S., Ishihara Y., Ozaki A., Hara Y., Harada Y., Mano Y., Sato T., Sato K., Sasaki K., Ishiguro H., Ohno S, & Akimoto K. (2020). High PKCλ expression is required for ALDH1-positive cancer stem cell function and indicates a poor clinical outcome in late-stage breast cancer patients. PLoS ONE, 15(7), e0235747.
+ Open protocol
+ Expand
7

Quantifying Cell Viability in HeLa Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
A HeLa cell suspension (0.5 × 105 cells/ml) was inoculated in 24-well plates 1 day before viral infection. Cells were collected in Trypsin-EDTA (Wako) and stained with 0.4% (w/v) Trypan Blue Solution (Wako). Cell counting was performed using a TC20 Automated Cell Counter (Bio-Rad), and the cell viabilities of virus-infected cells normalized to that of mock-treated cells were determined.
Takahashi T., Nakano Y., Onomoto K., Yoneyama M, & Ui-Tei K. (2019). LGP2 virus sensor enhances apoptosis by upregulating apoptosis regulatory genes through TRBP-bound miRNAs during viral infection. Nucleic Acids Research, 48(3), 1494-1507.
+ Open protocol
+ Expand
8

Cell Counting and Viability Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded at a density of 5 × 104 per well in a six-well plate and grown at 37°C in 5% CO2. The cells were harvested with 0.25% Trypsin-EDTA (Gibco, NY, USA) and mixed with an equal volume of 0.4% Trypan Blue solution (Wako, Osaka, Japan). Cell number and viability were determined using a hemocytometer. To calculate cell viability, at least 200 cells were counted in each culture after four days.
Sato H., Kato H., Yamaza H., Masuda K., Nguyen H.T., Pham T.T., Han X., Hirofuji Y, & Nonaka K. (2017). Engineering of Systematic Elimination of a Targeted Chromosome in Human Cells. BioMed Research International, 2017, 6037159.
+ Open protocol
+ Expand
9

Isolation and Culture of Primary Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Hygromycin B Gold was purchased from InvivoGen Co. (San Diego, CA, USA). HEPES was purchased from Dojindo Laboratories (Kumamoto, Japan). Hank's balanced salt solution was purchased from Sigma-Aldrich (St. Louis, MO, USA). Deoxyribonuclease I from bovine pancreas, precrystalline, DISPASE II, NP-40 substrate, tris (hydroxymethyl) aminomethane, sodium deoxycholate, sodium dodecyl sulfate, chloroform, isopropyl alcohol, ethanol, NaHCO3, d-(+)-glucose, and 0.4 w/v % trypan blue solution were purchased from FUJIFILM Wako Pure Chemical Co. (Osaka, Japan). Collagenase D was purchased from Roche Diagnostics, Inc. (Mannheim, Germany). Fetal bovine serum (FBS) was purchased from Biosera (East Sussex, UK). Dulbecco's modified Eagle's medium (DMEM) was purchased from Nissui Pharmaceutical Co. Ltd. (Tokyo, Japan). Mitomycin C, a penicillin-streptomycin-glutamine mixed solution, and 100 mM sodium pyruvate solution (100 × ) were purchased from Nacalai Tesque Inc. (Kyoto, Japan). All the other chemicals used were of the highest commercially available grade.
Tsujimura M., Kusamori K., Takamura K., Ito T., Kaya T., Shimizu K., Konishi S, & Nishikawa M. (2022). Quality evaluation of cell spheroids for transplantation by monitoring oxygen consumption using an on-chip electrochemical device. Biotechnology Reports, 36, e00766.
+ Open protocol
+ Expand
10

Cell Counting Using Trypan Blue

Check if the same lab product or an alternative is used in the 5 most similar protocols
To count the number of DT40 cells, 50 µl cultured medium was mixed with 0.4 wt/vol% Trypan Blue Solution (Wako), and the mixture was counted using Countess II (Thermo Fisher).
To count the number of RPE-1 cells, the culture medium was removed by aspirator. Then, 2.5 g/liter of Trypsin 1 mmol/liter EDTA solution (Nacalai Tesque) was added and incubated for 3 min at 37°C. To stop trypsinization, RPE-1 culture medium was added. The cell solution was mixed with same volume of 0.4 wt/vol% Trypan Blue Solution, and cell numbers were counted using Countess II.
Watanabe R., Hara M., Okumura E.I., Hervé S., Fachinetti D., Ariyoshi M, & Fukagawa T. (2019). CDK1-mediated CENP-C phosphorylation modulates CENP-A binding and mitotic kinetochore localization. The Journal of Cell Biology, 218(12), 4042-4062.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!