Cellometer auto x4

Manufactured by Revvity

Sourced in United States

The Cellometer Auto X4 is a compact and automated cell counter that provides accurate and reproducible cell counts. It utilizes advanced imaging technology to rapidly analyze cell samples and determine the total cell count and viability. The Cellometer Auto X4 is designed for efficient cell analysis in a wide range of applications.

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Lab products found in correlation

Whatman, by Cytiva (3 mentions) Gf c filter paper, by Cytiva (2 mentions) Lsr 2, by BD (1 mentions) Rat anti mouse ly 6g, by Thermo Fisher Scientific (1 mentions) Du730, by Beckman Coulter (1 mentions) Chemidoc system, by Bio-Rad (1 mentions) Dm6000b microscope, by Leica (1 mentions) Cellometer sd100, by Revvity (1 mentions) Uv 1800, by Shimadzu (1 mentions) Cellsens dimension v1, by Olympus (1 mentions) Ix73 microscope, by Olympus (1 mentions) Cytospin 4, by Thermo Fisher Scientific (1 mentions) 70 μm nylon cell strainer, by BD (1 mentions) Freund s adjuvant, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)

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Cellometer (154 citations) Cellometer Auto X4 Cell Counter (6 citations)

11 protocols using cellometer auto x4

Thoracic Lymph Collection and Analysis

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Mice were anesthetized with isoflurane (4% for induction, 2–3% for maintenance). The anesthetized animal was positioned on its right side and a cannula was inserted into the thoracic lymph duct above the cisterna chyli between the transverse lumbar artery and the diaphragm. Lymph was collected continuously, on average for 45 minutes to 1 hour, with a tube attached to a syringe coated with EDTA 0.1 M. Collected lymph was centrifuged at 1,200 g for 10 minutes at 4°C to remove most traces of whole cells, while being cautious not to create EVs due to high-speed centrifugation. Following that, 5% sucrose was added to each sample to improve preservation of the sample, which was then placed in a freezer at −80°C for further batch analysis. Although our technique ensures extreme care during collection, the absence of contaminating cells in lymph isolated from the thoracic duct was verified using a PC-based automated cell counter (Cellometer Auto X4, equipped with a 470/535 nm optic module; Nexcelom (Massachusetts (MA), USA)) that allowed bright field and fluorescent cell counting using acridine orange (AO), a nuclear-staining cell-permeable dye that stains all nucleated cells to generate green fluorescence. RBCs were considered as AO⁻ cells larger than 5 µm.

Milasan A., Tessandier N., Tan S., Brisson A., Boilard E, & Martel C. (2016). Extracellular vesicles are present in mouse lymph and their level differs in atherosclerosis. Journal of Extracellular Vesicles, 5, 10.3402/jev.v5.31427.

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Multicolor Flow Cytometry Analysis of Lung Immune Cells

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Cellularity was determined by flow cytometry using reagents obtained from BD Biosciences (San Jose, CA) unless otherwise specified. The resuspended BALF and MLN cells were divided in two for differential and intracellular cytokine staining. One half of the BALF cells were stained for 30 minutes using the following antibodies at 1∶100 rat anti-mouse Ly-6G, rat anti-mouse Siglec-F, pan-leukocyte rat anti-mouse CD45, and Armenian hamster anti-mouse CD11c (eBioscience Inc. San Diego, CA). MLN cells were also stained to identify B cell populations using B220/CD45R antibodies. After staining, cells were washed and fixed with BD Cytofix. Cells were then washed and resuspended in FACS buffer.
Cell populations were evaluated on a BD LSRII (BD Biosciences, San Jose, CA). Neutrophils were defined as CD45^hiLy-6G^hi, eosinophils as Ly-6G^lowSiglecF^hiCD11c^low, and alveolar macrophages as Ly-6G^lowSiglecF^hiCD11c^hi as previously reported [14] (link), [19] (link). Total cell numbers were quantified using acridine orange nuclear staining and an automated cell counter (Cellometer AutoX4, Nexcelom Bioscience, Lawrence, MA). Total numbers of each cell population were obtained by multiplying the frequency of specific population by the total number of BALF cells recovered for each animal.

Buskirk A.D., Green B.J., Lemons A.R., Nayak A.P., Goldsmith W.T., Kashon M.L., Anderson S.E., Hettick J.M., Templeton S.P., Germolec D.R, & Beezhold D.H. (2014). A Murine Inhalation Model to Characterize Pulmonary Exposure to Dry Aspergillus fumigatus Conidia. PLoS ONE, 9(10), e109855.

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Photosynthetic Pigment Quantification in Chlorella

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Photoautotrophically grown cells of the C. vulgaris wild type, E5 and the complemented strains were concentrated to 2 × 10⁸ cells/mL, and images were taken by a digital camera. After 15 min of dark adaptation, chlorophyll fluorescence images were taken by a ChemiDoc system (Bio-Rad, USA) with blue epi-illumination and a 695/55 nm filter.
The contents of chlorophyll a (Chl a), chlorophyll b (Chl b) and carotenoid (Car) were analyzed by a dimethyl sulfoxide (DMSO) extraction method^{59 (link)}. Briefly, an amount of 1 mL of cells in the mid-exponential growth phase was centrifuged at 13,000 rpm for 2 min. After the cell pellet was suspended in 1 mL DMSO, the mixture was incubated at 60 °C for 40 min and then centrifuged again. The optical density of the supernatant was measured at 480, 649 and 665 nm (OD₄₈₀, OD₆₄₉, OD₆₆₅) using a UV-Vis spectrophotometer (DU730; Beckman Coulter, Germany). The concentrations of the pigments were calculated by following equations:

\begin{matrix} C h l a [{μ g mL}^{- 1}] & = & 12.19 \times O D_{665} - 3.45 \times O D_{649} \\ C h l b [{μ g mL}^{- 1}] & = & 21.99 \times O D_{649} - 5.32 \times O D_{665} \\ C a r [{μ g mL}^{- 1}] & = & [1000 \times O D_{480} - 2.14 \times (C h l a) - 70.16 \times (C h l b)] / 220 \end{matrix}

The calculated concentrations were normalized with respect to the cell density of each sample, as determined by a Cellometer (Cellometer Auto X4, Nexcelom Bioscience, USA).

Shin W.S., Lee B., Kang N.K., Kim Y.U., Jeong W.J., Kwon J.H., Jeong B.R, & Chang Y.K. (2017). Complementation of a mutation in CpSRP43 causing partial truncation of light-harvesting chlorophyll antenna in Chlorella vulgaris. Scientific Reports, 7, 17929.

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Measuring Symbiodinium Cell Size

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The size of Symbiodinium cells was measured with an automated cell counter (Cellometer Auto X4; Nexcelom Bioscience, Lawrence, MA, USA). Symbiodinium cells were collected in the middle of the light period by centrifugation (16 000 g) for 1 min. The collected Symbiodinium cells were resuspended in fresh seawater and mixed by vortexing. After these steps, Symbiodinium cells lost their flagella and were all in the vegetative form (round cell shape without flagella). Measurements were conducted three times using separately cultured cells. Motile and vegetative Symbiodinium cells (Supplementary Figure S1) were photographed using a Leica DM6000B microscope (Leica Microsystems) equipped with a SPOT Flex camera (SPOT Imaging Solutions, Sterling Heights, MI, USA).

Biquand E., Okubo N., Aihara Y., Rolland V., Hayward D.C., Hatta M., Minagawa J., Maruyama T, & Takahashi S. (2017). Acceptable symbiont cell size differs among cnidarian species and may limit symbiont diversity. The ISME Journal, 11(7), 1702-1712.

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Synechocystis Colony Formation Assay

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Cultures of wild type and mutant Synechocystis strains were grown until they reached the exponential phase (O.D.₇₃₀ : 2.5); then, they were diluted 1:10 in fresh medium (O.D.₇₃₀ at day 0: 0.2). Each experiment was performed in triplicate i) under continuous white light (50 μE m⁻² s⁻¹), ii) with 12 h light/12 h dark cycles or iii) complete darkness. After five days, we assessed the cultures capabilities to generate colonies on plates: each liquid culture was diluted to an O.D.₇₃₀ between 0.3 and 0.4 and quantified using the cell counter Cellometer Auto X4 with a Cellometer SD100 counting chamber (Nexcelom Bioscience LLC.). The cell densities of the diluted cultures ranged between 1 × 10⁶ and 1.5 × 10⁶ cells/ml. Each diluted culture was then plated in three serial dilutions (1:2500, 1:5000 and 1:10000) and incubated at 30 °C in a 20 μE m⁻² s⁻¹ light regimen. Colonies, usually appeared after about ten days, were finally counted. Experiments were made in triplicate.

De Rosa E., Checchetto V., Franchin C., Bergantino E., Berto P., Szabò I., Giacometti G.M., Arrigoni G, & Costantini P. (2015). [NiFe]-hydrogenase is essential for cyanobacterium Synechocystis sp. PCC 6803 aerobic growth in the dark. Scientific Reports, 5, 12424.

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Photobioreactor Cultivation of Microalgae

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Wild-type and NsChlb transformants were cultivated under high light (200 μmol/m²/s) and mid light (90 μmol/m²/s) in a F2 N medium at 25 °C with 120 rpm agitation. Each baffled flask contained 200 mL of cells, and a carbon source was directly supplied into the flasks in the form of 2% CO₂ at 0.5vvm. The growth was determined by optical density, cell density, and dry cell weight (DCW). For each analysis, an automated cell counter (Cellometer^® Auto X4, Nexcelom, USA), UV–VIS spectrophotometer (UV-1800, Shimadzu, Japan), and GF/C filter paper (Whatman, USA) were used. The DCW was calculated by weighing the filter paper before and after filtering cells, which went through a delicate washing and drying procedure.

Koh H.G., Kang N.K., Jeon S., Shin S.E., Jeong B.R, & Chang Y.K. (2019). Heterologous synthesis of chlorophyll b in Nannochloropsis salina enhances growth and lipid production by increasing photosynthetic efficiency. Biotechnology for Biofuels, 12, 122.

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Analyzing Cell Growth Metrics

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During cultivation, cell growth was analyzed by referring to cell number and dry cell weight (DCW). For cell counting, 20 μl of culture was loaded onto Cellometer counting chambers (CHT4-002, Nexcelom, USA), which was then analyzed using an automated cell counter (Cellometer Auto X4, Nexcelom). The DCW was calculated by filtering the cells through GF/C filter paper (Whatman, USA), followed by washing with distilled water. The mass of the filter paper was measured before and after filtration under completely dry conditions.

Koh H.G., Jeong Y.T., Lee B, & Chang Y.K. (2021). Light Stress after Heterotrophic Cultivation Enhances Lutein and Biofuel Production from a Novel Algal Strain Scenedesmus obliquus ABC-009. Journal of Microbiology and Biotechnology, 32(3), 378-386.

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Cell Morphology and Viability Analysis

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Cell size, count and viability were determined using a Nexcelom Cellometer Auto X4 (Lawrence, MA) with Acridine Orange/Propidium Iodide staining solution (AO/PI, Nexcelom Bioscience). After day 14, cell count and viability were determined using a hemacytometer and trypan blue solution (Mediatech, Inc., Manassas, VA). All brightfield cell images were captured using an Olympus CK40 microscope (Center Valley, PA) with QCapture Pro v6.0 software (QImaging, Surrey, Canada), and images of stained slides were captured using an Olympus IX73 microscope with cellSens Dimension v1.12 software (Olympus). To measure enucleation frequency, cells were centrifuged onto slides using a Cytospin 4 (Thermo Scientific, Waltham, MA), fixed, stained with Wright-Giemsa, and visualized under 40x magnification. Three independent observers each counted 3 random fields of 100 cells, in a blinded fashion, to obtain an average value. Enucleation frequency was calculated by dividing the number of enucleated cells by the total number of cells.

Shah S.N., Gelderman M.P., Lewis E.M., Farrel J., Wood F., Strader M.B., Alayash A.I, & Vostal J.G. (2016). Evaluation of Stem Cell-Derived Red Blood Cells as a Transfusion Product Using a Novel Animal Model. PLoS ONE, 11(12), e0166657.

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Measuring C. sorokiniana HS1 Growth

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Cell growth of C. sorokiniana HS1 in the broth was analyzed by measuring dry cell weight and cell density. In order to determine dry cell weight (DCW), samples were passed through circular glass filter paper (CF/G, 47 mm, Whatman), which was washed with deionized water (DW), and dried in a 70 °C oven overnight. For counting cells, an automated cell counter was used (Cellometer auto X4, Nexcelom, USA).

Oh S.H., Chang Y.K, & Lee J.H. (2019). Identification of significant proxy variable for the physiological status affecting salt stress-induced lipid accumulation in Chlorella sorokiniana HS1. Biotechnology for Biofuels, 12, 242.

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Adoptive Transfer of OTII and Ig-Tg Cells

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Spleens were harvested from male donor OTII mice and pressed through 70 μm nylon cell strainers (Falcon) in DMEM (Cellgro) supplemented with 2% FBS (Atlanta Biologicals), 10 mM HEPES, 50 IU/mL of penicillin, and 50 μg/mL of streptomycin (HyClone). Splenocytes were centrifuged for 7 minutes at 380 rcf., 4°C and resuspended in 0.14 M NH₄Cl in 0.017 M Tris buffer, pH 7.2 for erythrocyte lysis, washed twice with DMEM supplemented as above, and counted using a Cellometer Auto X4 (Nexcelom). The fraction of CD19⁻ CD8⁻ CD4⁺ Vβ5⁺ (OTII) splenocytes was determined by flow cytometry, and the indicated number of OTII cells were transferred i.v. to male recipient mice. Ig-Tg B cells were enriched from male or female donor mice by negative selection as previously described (Allen et al., 2007 (link)). For transient exposure to Ag, purified Ig-Tg B cells were incubated with the indicated concentration of DEL-OVA ex vivo for 5 minutes at 37°C, washed four times with DMEM supplemented as above, and transferred i.v. to recipient mice. Where indicated, recipient mice were immunized s.c. in the flanks and base of tail with 50 or 5 μg of the indicated Ag emulsified in complete or incomplete Freund’s adjuvant (Sigma), prepared according to the manufacturer’s directions. Where indicated, recipient mice were injected s.c. in the base of tail with unconjugated αDEC-205, αDEC-OVAp or iso-OVAp in PBS.

Turner J.S., Ke F, & Grigorova I.L. (2018). B Cell Receptor Crosslinking Augments Germinal Center B Cell Selection when T Cell Help Is Limiting. Cell reports, 25(6), 1395-1403.e4.

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