Canto 1 flow cytometer

Manufactured by BD

Sourced in United States

The Canto I flow cytometer is a compact and versatile instrument used for the analysis of cells and particles in liquid samples. It is designed to detect and measure multiple parameters of individual cells or particles as they pass through a focused laser beam. The Canto I is capable of analyzing a wide range of sample types, including cells, microbeads, and other particles, providing researchers and clinicians with valuable data for their applications.

Automatically generated - may contain errors

Lab products found in correlation

4 protocols using canto 1 flow cytometer

Comprehensive Blood Cell Phenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols

The phenotypes of cells in the blood were analyzed by flow cytometry according to standard procedures at the Hematopathology Unit, Dept. of Pathology, Karolinska University Hospital, using 6 color flow cytometry to detect T, B, NK cells and CD3− CD4+ cells (monocytes and dendritic cells). Flow cytometry was performed on a CANTO 1 flow cytometer (BD, Becton-Dickinson, Europe).
For data acquisition and analysis, a CANTO 1 flow cytometer (BD, Becton Dickinson, Europe) was used with Cell Quest software (Becton Dickinson, Franklin Lakes, New Jersey, USA). All samples were analyzed by setting appropriate side and forward scatter gates to identify the mononuclear cell population, using CD45 and forward and side scatter for gate setting. Consistency of analysis parameters was ascertained by calibrating the flow cytometer with calibrating beads and FacsComp software, both from Becton Dickinson. The results are reported as percentage of gated cells positive for each antibody. The following fluorochrome conjugated antibodies, all from BD, were used: CD4 PE, CD3 PerCP-Cy5.5, CD19 PE-Cy7, CD8 APC and CD45 APC-H7. We also used BD Multitest 6-Color TBNK Reagent containing CD3 FITC clone SK7, CD16 PE clone B73, CD56 PE clone NCAM 16.2, CD45 PerCP-Cy5.5 clone 2D1, CD4 PE-Cy7 clone SK3, CD19 APC clone SJ25C1 and CD8 APC-Cy7, clone SK1. The gating strategy is shown in Fig. S1.

Almestrand S., Wang X., Jeppsson-Ahlberg Å., Nordgren M., Flygare J., Christensson B., Rössner S, & Sander B. (2015). Influence of rimonabant treatment on peripheral blood mononuclear cells; flow cytometry analysis and gene expression profiling. PeerJ, 3, e1056.

+ Open protocol

+ Expand

Murine Immune Cell Isolation and Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Single-cell suspensions were prepared from spleen and blood. Erythrocytes were depleted by lysis with NH₄Cl₂ solution. Peritoneal fluid was collected after lavage with cold and sterile PBS. Murine cells were harvested and cultured in Roswell Park Memorial Institute (RPMI) media with 10% foetal calf serum (FCS) plus supplement (glutamine, 2-mercaptoethanol, penicillin and streptomycin, and 10 mM HEPES). For cell-surface staining, 2–3 × 10⁶ cells per sample were incubated with various antibodies in staining buffer (PBS and 3% FCS) for 20 min at 4 °C. After 4 h of incubation in media with PMA (50 ng/ml) and ionomycin (500 ng/ml) intracellular cytokine staining was performed according to the manufacturer’s guidelines for the BD Biosciences-Pharmingen Fixation/Permeabilization Solution Kit. Anti-murine antibodies included B220 (RA3-6B2), CD4 (GK1.5), CD8 (53-6.7), CD69 (H1.2F3), CD62L (MEL-14), CD44 (IM7), Gr-1 (RB6-8C5), CD11b (M1/70), CD11c (N418), F4/80 (BM8), Ly6C (HK1.4), MHCII (M5/114.15.2) and IFNγ (XMG1.2) all from eBioscience (San Diego, USA). Data were acquired on a Canto I flow cytometer (BD Biosciences, New Jersey, USA) and analysed using FlowJo for Mac version 9.2 (Tree Star Inc., Ashland, USA).

Staats K.A., Humblet-Baron S., Bento-Abreu A., Scheveneels W., Nikolaou A., Deckers K., Lemmens R., Goris A., Van Ginderachter J.A., Van Damme P., Hisatsune C., Mikoshiba K., Liston A., Robberecht W, & Van Den Bosch L. (2016). Genetic ablation of IP3 receptor 2 increases cytokines and decreases survival of SOD1G93A mice. Human Molecular Genetics, 25(16), 3491-3499.

+ Open protocol

+ Expand

Mitochondrial Mass and DNA Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mitochondrial mass was measured using flow cytometry after MitoTracker Green staining of R1(LL) and R1(SS) mESCs before or after overnight starvation. Briefly, cell were washed with PBS and incubated 50 nM of MitoTracker green for 30 min before analysis on a Canto I flow cytometer (BD Biosciences). Data analysis was performed using the FlowJo software (Tree Star, Ashland, OR, USA).
Mitochondrial DNA amount was evaluated by measuring the ratio of mitochondrial DNA (mtDNA) versus nuclear DNA (nDNA) by qPCR analysis. Briefly, the DNA of R1(LL) and R1(SS) mESCs after overnight starvation was isolated using DNAzol (Invitrogen) following manufacturer’s protocol. mtDNA was measured using mt-Co1 primers mt-Co1-F 5⁰-CAGTCTAATGCTTACTCAGC-3⁰ and mt-Co1-R 5⁰-GGGCAGTTACGATAACATTG-3⁰, and nDNA was measured using Gapdh primers Gapdh-F 5⁰GGGAA GCCCATCACCATCTTC-3⁰ and Gapdh-R 5⁰AGAGGGGCCATCCACAGTCT-3⁰. Each reaction contained 10ng of DNA extract, 13 SYBR Green Master Mix, and 300nM of each primer. The qPCR was performed using a 7300 real-time PCR system (Applied Biosystems) and the ratios mtDNA/nDNA were measured.

Hussein A.M., Wang Y., Mathieu J., Margaretha L., Song C., Jones D.C., Cavanaugh C., Miklas J.W., Mahen E., Showalter M.R., Ruzzo W.L., Fiehn O., Ware C.B., Blau C.A, & Ruohola-Baker H. (2020). Metabolic Control over mTOR-Dependent Diapause-like State. Developmental cell, 52(2), 236-250.e7.

+ Open protocol

+ Expand

Phenotypic Characterization of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells (1 × 10 5 /100 L) were stained for 30 min at 4 • C with the following anti-human antibodies at the appropriate concentration or with the relevant isotypes: CD83-FITC (BD Biosciences), CD14-PE (Beckman coulter), CD86-PE (BD Biosciences), HLA-DR-APC (BD Biosciences), CD207-APC (Biolegend), CD1A-AF488 (Biolegend), CD123-APC (Biolegend), BDCA-2-APC (Biolegend), CD209-PE (Beckman coulter). The cells were then washed with 1 × PBS (Dutscher) and viable cells analysed on a Canto I flow cytometer (BD Biosciences). Results were expressed as the ratio of MFI (mean of fluorescence) of the marker on the MFI of the isotype control and referred as MFI ratio.

Deluce-Kakwata-Nkor N., Lamendour L., Chabot V., Héraud A., Ivanovic Z., Halary F., Dehaut F, & Velge-Roussel F. (2018). Differentiation of human dendritic cell subsets for immune tolerance induction. Transfusion clinique et biologique : journal de la Societe francaise de transfusion sanguine, 25(1).

+ 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?

Revolutionizing how scientists
search and build protocols!