Sa3800 spectral cell analyzer

Manufactured by Sony

Sourced in Japan, United States

The SA3800 Spectral Cell Analyzer is a laboratory instrument designed for the analysis of cells and particles. It utilizes spectroscopic techniques to provide detailed information about the properties and characteristics of the samples under investigation.

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Lab products found in correlation

Sh800 cell sorter, by Sony (4 mentions) Sepmate tube, by STEMCELL (3 mentions) Biotinylated pd 1 and pe streptavidin, by BioLegend (2 mentions) Alexa fluor 488 conjugated anti mouse igg, by Cell Signaling Technology (2 mentions) B cell isolation kit, by Miltenyi Biotec (2 mentions) Pam3csk4, by InvivoGen (2 mentions) Facsdiva software version 8, by BD (1 mentions) Facsaria fusion instrument, by BD (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Accumax, by STEMCELL (1 mentions) Aldefluor kit, by STEMCELL (1 mentions) D9542, by Merck Group (1 mentions) Fitc annexin 5 apoptosis detection kit, by BD (1 mentions) Dcfh da, by Dojindo Laboratories (1 mentions) Stempro accutase cell dissociation reagent, by Thermo Fisher Scientific (1 mentions) Aldefluor assay kit, by STEMCELL (1 mentions) Alphalisa kit, by PerkinElmer (1 mentions) Cytoflex lx cytometer, by Beckman Coulter (1 mentions) Flow cytometry, by BD (1 mentions) Special order cytometer, by BD (1 mentions)

Close spelling variants of this product

SA3800 Spectral Analyzer (39 citations) SA3800 (36 citations) SA3800 Cell Analyzer (33 citations) Spectral Analyzer (8 citations) Spectral Cell Analyzer (3 citations) SA3800 analyzer (2 citations)

37 protocols using sa3800 spectral cell analyzer

Staining and Analysis of Kidney Organoids

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Kidney organoids were stained with fluorescein-conjugated LTL (FL-1321, Vector Laboratories) as described elsewhere (Garreta et al., 2019 (link)). Kidney organoids were then dissociated to single cells using Accumax (07921, Stem Cell Technologies) for 15 min followed by 0.25% (wt/vol) trypsin (25300–054, Life Technologies) for 15 min at 37 °C. SA3800 software version 2.0.4 (SONY) was used to acquire flow cytometry samples in the Sony SA3800 spectral cell analyzer (SONY). FACSDiva software version 8.0.1 (BD Biosciences) was used in the FACS Aria Fusion instrument (BD Biosciences) for cell sorting experiments. FlowJo software version 10 was used to analyze the data.

Garreta E., Prado P., Stanifer M.L., Monteil V., Marco A., Ullate-Agote A., Moya-Rull D., Vilas-Zornoza A., Tarantino C., Romero J.P., Jonsson G., Oria R., Leopoldi A., Hagelkruys A., Gallo M., González F., Domingo-Pedrol P., Gavaldà A., del Pozo C.H., Hasan Ali O., Ventura-Aguiar P., Campistol J.M., Prosper F., Mirazimi A., Boulant S., Penninger J.M, & Montserrat N. (2022). A diabetic milieu increases ACE2 expression and cellular susceptibility to SARS-CoV-2 infections in human kidney organoids and patient cells. Cell Metabolism, 34(6), 857-873.e9.

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Evaluation of SI-2 Inhibitor Effects

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For each analysis, THJ-11T and -16T cells were first incubated with the indicated dose of SI-2 or vehicle for 24 or 48 hours. For cell cycle analysis, the cells were trypsinized, washed, and fixed in 70% ethanol at – 20°C overnight. On the next day, the cells were washed and then incubated in DAPI staining solution (D9542, Sigma-Aldrich: 1 µg/ml DAPI in PBS) for 10 minutes at room temperature. Cell apoptosis was measured with Annexin V-FITC apoptosis detection kit (556547, BD Biosciences). To assay for ALDH activity, we used an ALDEFLUOR kit (#01700, Stem Cell Technology, Lt.,) as per the manufacturer’s protocol. Briefly, the cells were trypsinized, after which 1 million cells were resuspended in 1 ml aldeflour assay buffer. Aldeflour reagent was added to this cell suspension and then half of the cell suspension was transferred to a fresh tube containing diethylaminobenzaldehyde (DEAB), which is used to control for background fluorescence as a specific inhibitor of ALDH. The cells were incubated at 37°C for 30 minutes, washed, and then subjected to flow cytometry. All the stained cells were analyzed by Sony SA3800 spectral cell analyzer (Sony Biotechnology, Inc.). The data analysis was done by FlowJo software 10.5.3 (FlowJo LLC).

Lee W.K., Kim W.G., Fozzatti L., Park S., Zhao L., Willingham M.C., Lonard D., O’Malley B.W, & Cheng S.Y. (2020). Steroid receptor coactivator-3 as a target for anaplastic thyroid cancer. Endocrine-related cancer, 27(4), 209-220.

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Quantifying Cellular and Mitochondrial ROS

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Two methods were used to determine ROS production. Mitochondrial ROS was determined with MitoSOX stain as we reported 25, 26 . To detect cellular ROS, cultured cells at 80% confluency were treated with DZ-SIM (5 μM) for 6 hours. The cells (1 × 10 7 ) were trypsinized and collected in 1 ml of fresh medium and were stained with diacetyldichlorofluorescein diacetate (DCFH-DA, Dojindo Laboratories, Rockville, MD, USA), with the manufacturer's recommended protocol. After being washed in phosphate buffered saline twice, the samples were subjected to flow cytometry on a Sony SA3800 spectral cell analyzer (Sony Biotechnology, San Jose, CA, USA).

Ou Y., Chu G.C., Lyu J., Yin L., Lim A., Zhai N., Cui X., Lewis M.S., Edderkaoui M., Pandol S.J., Wang R, & Zhang Y. (2024). Overcoming Resistance in Prostate Cancer Therapy Using a DZ-Simvastatin Conjugate. Molecular pharmaceutics, 21(2).

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Cell Dissociation and Flow Cytometry

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Differentiated cells were dissociated by treatment with StemPro Accutase Cell Dissociation Reagent (Thermo Fisher Scientific) for 8–10 min. Collected cells were fixed, permeabilized, blocked, and incubated with antibody as mentioned in section 2.6. The antibodies used in this study are summarized in Supplemental Table 2. The analysis was performed using SONY SA3800 spectral cell analyzer (Sony Biotechnology, San Jose, CA, USA) (see also Supplemental Fig. 2A).

Sekiya A., Takasawa K., Arai Y., Horike S.I., Akutsu H., Umezawa A, & Nishino K. (2022). Variation of DNA methylation on the IRX1/2 genes is responsible for the neural differentiation propensity in human induced pluripotent stem cells. Regenerative Therapy, 21, 620-630.

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Quantification of GD2 Antigen Expression

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The expression of GD2 antigen on U87MG and U87DR surface was analyzed by flow cytometry analysis using anti-GD2 Ab (14G2a). To perform this assay, cells were trypsinized and detached from the culture plates. The cells were collected and washed with PBS for incubation with unlabeled anti-GD2 Ab (1 μg per 10⁶ cells) for 1 h. Followed by this, the cells were washed in PBS containing 1%FBS and 0.02% sodium azide. Subsequently, the cells were incubated with fluorescein isothiocyanate (FITC)-labeled anti-mouse IgG (1:1000) for 40 min and washed twice in PBS. The samples were then analyzed using FACS instrument (SONY SA3800 spectral cell analyzer) where 10,000 events were collected for each sample. The data so obtained were further analyzed using the FlowJo software to determine the extent of GD2 expression.

Jose G., Lu Y.J., Hung J.T., Yu A.L, & Chen J.P. (2020). Co-Delivery of CPT-11 and Panobinostat with Anti-GD2 Antibody Conjugated Immunoliposomes for Targeted Combination Chemotherapy. Cancers, 12(11), 3211.

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Quantifying ALDH Activity in Cells

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ALDH activity of various cell lines was quantified using an ALDEFLUOR^® assay kit (STEMCELL Technologies), according to the manufacturer's instructions. Cells were harvested, placed in assay buffer, and incubated for 45 min at 37°C to allow intracellular ALDH to convert uncharged ALDH substrate (BAAA) to negatively charged BAA. As a negative control, an aliquot of ALDEFLUOR-stained cells was quenched immediately with 1.5 mM diethylaminobenzaldehyde (DEAB), a specific ALDH inhibitor. Cells were analyzed using the green fluorescence channel (FL1) on a SA3800 spectral cell analyzer (Sony Biotechnology).

Liang Y.J., Wang C.Y., Wang I.A., Chen Y.W., Li L.T., Lin C.Y., Ho M.Y., Chou T.L., Wang Y.H., Chiou S.P., Lin Y.J, & Yu J. (2017). Interaction of glycosphingolipids GD3 and GD2 with growth factor receptors maintains breast cancer stem cell phenotype. Oncotarget, 8(29), 47454-47473.

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EV Effects on Immune Cells

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PBMCs were isolated from whole blood using SepMate tubes (STEMCELL Technologies). Cells were plated in round-bottom 96-well plates at 200,000 cells per well in RPMI supplemented with 10% fetal bovine serum. Different EVs were added to the wells in a final volume of 200 µL and incubated overnight at 37 °C and 5% CO₂. The next day, the cells were pelleted, washed, and stained for flow cytometry. Level of the GFP was assessed, along with different population markers (CD3—T Cells, CD19—B Cells, CD16/56—NK Cells, CD14—Monocytes, CD123—pDCs, CD11c—cDCs). Flow cytometry analysis was completed on a SA3800 Spectral Cell Analyzer (Sony).

Jang S.C., Economides K.D., Moniz R.J., Sia C.L., Lewis N., McCoy C., Zi T., Zhang K., Harrison R.A., Lim J., Dey J., Grenley M., Kirwin K., Ross N.L., Bourdeau R., Villiger-Oberbek A., Estes S., Xu K., Sanchez-Salazar J., Dooley K., Dahlberg W.K., Williams D.E, & Sathyanarayanan S. (2021). ExoSTING, an extracellular vesicle loaded with STING agonists, promotes tumor immune surveillance. Communications Biology, 4, 497.

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Evaluating STING Agonist Efficacy in Human PBMCs

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Healthy human whole bloods were purchased from Research Blood Components LLC. All donors filled out their standard medical questionnaire and agreed to their donor informed consent form before blood donation. PBMCs were isolated from whole blood using SepMate tubes (STEMCELL Technologies). Cells were plated in round-bottom 96-well plates at 200,000 cells per well in RPMI supplemented with 10% fetal bovine serum. ExoSTING or free CDNs were added to the wells in a final volume of 200 µL and incubated overnight at 37 °C and 5% CO₂. The next day, the supernatant was harvested and analyzed for human IFN-β using an AlphaLISA kit according to the manufacturer’s protocol (Perkin Elmer). The cells were pelleted, washed, and stained for flow cytometry. Expression of the activation marker CD86 on CD11c+ dendritic cells and CD14+ monocytes were assessed. Flow cytometry analysis was completed either on a SA3800 Spectral Cell Analyzer (Sony) or Beckman Coulter CytoFLEX LX cytometer. EC₅₀ values were analyzed by using GraphPad Prism 8. Antibody information is listed in Supplementary Table 2.

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PD-L1 and HLA-G Expression on NSCLC Cells

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NSCLC cells were seeded into 12‐well plates and treated with chemotherapeutics for 48 h. Cells were then stained with FITC‐conjugated anti‐PD‐L1 or anti‐HLA‐G antibodies. The induction of PD‐L1 and HLA‐G expression was determined using flow cytometry (BD Bioscience). The MFI was analyzed using FlowJo™ software (FlowJo, LLC/BD Bioscience). The phenotypes and activating status of human PBMC‐derived T cells were assessed and analyzed using flow cytometry (SONY SA3800 spectral cell analyzer) after staining with specific antibodies.

Chen M., Hung M., Pan C., Huang S., Jan C., Li Y., Chiu S, & Cho D. (2023). Pemetrexed combined with dual immune checkpoint blockade enhances cytotoxic T lymphocytes against lung cancer. Cancer Science, 114(7), 2761-2773.

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Cell Cycle Analysis by Flow Cytometry

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Cells were harvested and washed with PBS, followed by fixation with 70% cold ethanol overnight at 4°C. Cells were washed twice with PBS and stained with propidium iodide. Cell cycle distribution was detected by SONY SA3800 spectral cell analyzer. Data were analyzed by FlowJo 7.6 software (Tree Star).

Shi X., Zheng Y., Jiang L., Zhou B., Yang W., Li L., Ding L., Huang M., Gery S., Lin D.C, & Koeffler H.P. (2020). EWS-FLI1 regulates and cooperates with core regulatory circuitry in Ewing sarcoma. Nucleic Acids Research, 48(20), 11434-11451.

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