Spectramax minimax 300 imaging cytometer

Manufactured by Molecular Devices

Sourced in United States

The SpectraMax MiniMax 300 Imaging Cytometer is a compact and versatile instrument designed for high-content imaging and cellular analysis. It features a motorized stage, autofocus capabilities, and a sensitive camera for capturing detailed images of cells and other biological samples.

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Lab products found in correlation

Celltiter glo luminescent cell viability assay, by Promega (2 mentions) Mtt assay, by Biotium (2 mentions) Resazurin dye, by Merck Group (2 mentions) Xf cell mito stress test kit, by Agilent Technologies (2 mentions) Xf96 cell culture microplate, by Agilent Technologies (2 mentions) Nucleospin plant 2 molecular kit, by Macherey-Nagel (2 mentions) Celleventtm caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions) Eclipse ti microscope, by Nikon (1 mentions) Softmax pro 7, by Molecular Devices (1 mentions) Dual luciferase reporter assay system, by Promega (1 mentions) Amplex ultrared reagent, by Thermo Fisher Scientific (1 mentions) Piercetm nickel coated plates, by Thermo Fisher Scientific (1 mentions) Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions) Seaprep agarose, by Lonza (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions) D300e digital dispenser, by Tecan (1 mentions) Fluoroskan ascent fl, by Thermo Fisher Scientific (1 mentions) Medetomidine, by Pfizer (1 mentions) Midazolam, by Ratiopharm (1 mentions)

20 protocols using spectramax minimax 300 imaging cytometer

Measuring Anchorage-Dependent and -Independent Cell Growth

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To measure anchorage-dependent growth, cells were plated at low density in 96 well plates and allowed to attach overnight. Inhibitors were then added with a D300e Digital Dispenser (Tecan). After 72 hours, cells were labeled with calcein AM (Thermo Fisher) according to the manufacturer’s protocol, and counted on a SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices).
To measure anchorage-independent growth, soft agar colony formation assays were performed. In a 96-well plate, wells were coated with 50 μl 2% SeaPrep Agarose (Lonza) mixed with cell culture medium. Next, 100 μl of 5000 cells mixed with 1% SeaPrep Agarose were added to each well and allowed to solidify. Fifty μl of medium were added to each well, and drugs were dispensed with a D300e Digital Dispenser (Tecan). After 72 hours, the alamarBlue cell viability reagent (Thermo Fisher) was added and incubated for 3 hours. Fluorescence was quantified on a SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices).

Blake D.R., Vaseva A.V., Hodge R.G., Kline M.P., Gilbert T.S., Tyagi V., Huang D., Whiten G.C., Larson J.E., Wang X., Pearce K.H., Herring L.E., Graves L.M., Frye S.V., Emanuele M.J., Cox A.D, & Der C.J. (2019). Application of a MYC degradation screen identifies sensitivity to CDK9 inhibitors in KRAS-mutant pancreatic cancer. Science signaling, 12(590), eaav7259.

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Apoptosis Assay for Breast Cancer Cells

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The T47Darom and T47DaromLR cells were seeded at a density of 1.5 × 10³ cells in a 96-well plate with appropriate media overnight. Cells were treated the following day with DMSO control, 0.5 μM lapatinib, 10 μM glyceollin, or 0.5 μM lapatinib plus 10 μM glyceollin for 5 days. Following the drug treatment, the cells were treated with 4 μM CellEventTM Caspase-3/7 Green Detection Reagent (Invitrogen, Carlsbad, CA, USA) and incubated for 30 min at 37 °C with 5% CO₂. The cells were then fixed with 4% formaldehyde and allowed to incubate for 15 min at room temperature. The SpectraMax^® MiniMax^® 300 Imaging Cytometer (Molecular Devices, San Jose, CA, USA) was used to measure fluorescence and background wavelengths at 530 nm and 502 nm, respectively. Cells were imaged on Nikon Ti Eclipse microscope (Nikon, Minato, Tokyo, Japan) using an FITC filter.

Walker R.R., Patel J.R., Gupta A., Davidson A.M., Williams C.C., Payton-Stewart F., Boué S.M., Burow M.E., Khupse R, & Tilghman S.L. (2022). Glyceollins Trigger Anti-Proliferative Effects in Hormone-Dependent Aromatase-Inhibitor-Resistant Breast Cancer Cells through the Induction of Apoptosis. International Journal of Molecular Sciences, 23(5), 2887.

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Proliferation Assay for Cancer Cells

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Proliferation was measured with the CellTiter-Glo Luminescent Cell Viability Assay (Promega, Madison, WI, USA) following the manufacturer’s protocol. SC cells were seeded with complete medium then treated with MMC or 5-FU the next day. Media with drug were replaced every 48 hours. Luminescence measurements were taken every 24 hours on the SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices). The mean percent luminance change compared to the baseline was graphed. Each treatment was performed in triplicate. Growth curves were analyzed by calculating the area under the curves (AUCs), which provide estimates of percent inhibition throughout the entire experiment. Ordinary one-way ANOVAs with Dunnett’s post-hoc comparison tests were performed to determine differences between the control and treated groups.

Gallo R.A., Lang S.H., Gomez A., Sabater A.L., Tse D.T., Pelaez D, & Rong A.J. (2022). Effects of mitomycin-C and 5-fluorouracil on ocular adnexal sebaceous carcinoma cells. American journal of ophthalmology, 240, 14-22.

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LPS, MPLA, and polyI:C Modulation of PyMT Tumor Cell Viability

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PyMT tumor cells were seeded to a 96-well plate
(5×10³/well) and treated with LPS (100 ng/ml), MPLA
(100 ng/ml), or polyI:C (1 μg/ml) with or without mouse IFNγ
(33 ng/ml) for 48 hours. Next, 20 μl CellTiter 96® AQueous One
Solution reagent was added to 100 μl medium and incubated for 2 hours
at 37 °C. Absorbance at 490 nm was measured using a SpectraMax
MiniMax 300 Imaging Cytometer (Molecular Devices).

Sun L., Kees T., Almeida A.S., Liu B., He X.Y., Ng D., Han X., Spector D.L., McNeish I.A., Gimotty P., Adams S, & Egeblad M. (2021). Activating a collaborative innate-adaptive immune response to control metastasis. Cancer cell, 39(10), 1361-1374.e9.

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Quantifying Hydrogen Peroxide with Amplex Red

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An Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit was used for the H₂O₂ concentration measurement, following the manufacturer’s instruction with modifications. Hydrogen peroxide standards and samples (10–50 µL) were loaded in triplicate into a 96 well black polystyrene microplate with a clear bottom (Corning). A working solution was immediately added for a final volume of 100 µL after each triplicate loading. The final concentration of horseradish peroxidase in the mixture was 0.1 U/mL, and Amplex Red was 0.05 mM. After 5–10 min incubation at room temperature in the dark, the plate was read with a microplate reader, SpectraMax MiniMax 300 Imaging Cytometer, equipped with SoftMax Pro 7 software (Molecular Devices). Fluorescence detection was set at λ_ex 540 nm and λ_em 580 nm and optical absorbance detection at 560 nm. Sample concentrations were calculated according to standard curves included in each experiment.

Guarino V.A., Oldham W.M., Loscalzo J, & Zhang Y.Y. (2019). Reaction rate of pyruvate and hydrogen peroxide: assessing antioxidant capacity of pyruvate under biological conditions. Scientific Reports, 9, 19568.

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MTT Assay for Cell Viability

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SC and LSC viabilities after 72-hour treatment with MMC or 5-FU were assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Biotium, Fremont, CA, USA) according to the manufacturer’s protocol, as previously described.^{20 (link)} Optical density was quantified with a SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices, San Jose, CA, USA) at 570 nm, and background absorbance was measured at 630 nm. Quadruplicate readings were recorded. Values were normalized to the untreated control. The half-maximal effective doses (EC₅₀) on cell viability were determined with Combenefit software.^{22 (link)}

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Quantitative Evaluation of DNA Repair Pathways

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Three reciprocal parental crosses were set up between the trans-heterozygous indel mutant (ku80^−/−, lig4^−/−, or DNA-PKcs^−/−) and Lvp strains; 1) ♂ gene^−/− x ♀ gene^−/−; 2) ♂ Lvp × ♀ gene^−/−; 3) ♂ Lvp × ♀ Lvp. Eight replicates (n = 50 embryos per each) of each embryo group were microinjected with the luciferase reporter plasmid for either NHEJ or SSA assay (pNHEJ2.1 or pSSA, 0.5 μg/μl), together with a normalization control plasmid (pSLfa-PUb-Renilla, 0.2 μg/μl) and a plasmid coding for the homing endonuclease (pY2-I-AniI, 0.2 μg/μl). For negative controls, pY2-I-AniI was replaced by a blank plasmid (pSLfa-mcs, 0.2 μg/μl) for microinjections. All injected embryos were incubated at 27°C in a humidity chamber for 48 hr post-injection, and the collected embryos were kept at −80°C until further analysis. The DNA repair event-dependent luciferase activity in each embryo group was measured by using the Dual-Luciferase Reporter Assay System (Promega) and the SpectraMax MiniMax 300 Imaging Cytometer (Molecular Devices).

Chae K., Overcash J.M., Dawson C., Valentin C., Tsujimoto H., Myles K.M, & Adelman Z.N. (2023). CRISPR-based gene editing of non-homologous end joining factors biases DNA repair pathway choice toward single-strand annealing in Aedes aegypti. Current research in biotechnology, 5, 100133.

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Nickel-Based Affinity Purification and Detection

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Cells (grown in 24 well plates) were lysed in lysis buffer (50 mM Tris-Hcl, 1M NaCl, 10 mM TCEP, 25 mM Imidazole, protease inhibitors) and processed as described above. Total protein (50 ug) was added to a single well on the Pierce^TM Nickel Coated Plates (Thermo Scientific) and incubated for 4–6 h at 4C. Each well was then gently washed 3 times with wash buffer (50 mM Tris-Hcl, 0.1M NaCl, 10 mM TCEP, 50 mM Imidazole). Each well was incubated with the anti-FLAG-M2-peroxidase antibody for 1 h at 4C. After washed each well 6 times with ice-cold PBS, the Amplex® UltraRed Reagent (Molecular Probes) assay solution was added (200 ul/well). Plates were incubated for 10-20 min at room temperature (in dark). 200 ul of the reaction was transferred to a 96well BLACK plate and read on the SpectraMax Minimax 300 Imaging Cytometer (Molecular Devices).

Ramachandran S., Coffin S.L., Tang T.Y., Jobaliya C.D., Spengler R.M, & Davidson B.L. (2016). Cis-acting single nucleotide polymorphisms alter MicroRNA-mediated regulation of human brain-expressed transcripts. Human Molecular Genetics, 25(22), 4939-4950.

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Caspase-3/7 Activation Assay in T47D Cells

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The T47Darom and T47DaromLR cells were seeded at a density of 1.5×103 cells in a 96-well plate with appropriate media overnight. Cells were treated the following day with DMSO (control) or 10 μM SB205380. Cells were incubated for 24 h at 37°C with 5% CO₂. On the following day, the cells treated with 4 μM CellEvent™ Caspase-3/7 Green Detection Reagent (Invitrogen) and incubated for 30 min at 37°C with 5% CO₂. The cells were then fixed with 4% formaldehyde and allowed to incubate for 15 min at room temperature. The SpectraMax®MiniMax® 300 Imaging Cytometer (Molecular Devices) was used to measure fluorescence and background wavelengths at 530 nm and 502 nm, respectively. Cells were imaged on Nikon Ti Eclipse microscope (Minato, Tokyo, Japan) using a FITC filter.

WALKER R.R., GALLEGOS K.M., BRATTON M.R., LEMIEUX K.P., ZHANG K., WANG G., DAVIDSON A.M, & TILGHMAN S.L. (2021). Acquisition of Letrozole Resistance Through Activation of the p38/MAPK Signaling Cascade. Anticancer research, 41(2), 583-599.

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Quantification of FITC-dextran Bioavailability

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Water bottles were removed from the cages 4 h before administration of 50 mg FITC-dextran (MW 4000) (cat. no. 46944, Sigma-Aldrich) per 100 g body weight in PBS. After 4 h, the mice were anesthetized by intraperitoneal injection of a solution of 5.0 mg kg⁻¹ body weight midazolam (Ratiopharm), 0.5 mg kg⁻¹ body weight medetomidine (Pfizer) and 0.05 mg kg⁻¹ body weight fentanyl (CuraMed Pharma) in 0.9% NaCl solution. Therefore, whole blood without anticoagulants was collected by cardiac puncture^{59 (link)} and stood overnight at 4 °C to coagulate. The next morning, serum was obtained by centrifugation (6,500 r.p.m., 10 min, 4 °C) of whole blood, aliquoted and stored at −20 °C for further analyses. The serum samples were diluted 1:1 (v/v) with PBS and measured in duplicate by spectrofluorometry (Fluoroskan Ascent FL, Thermo Fisher or SpectraMax MiniMax 300 Imaging CYtometer, Molecular Devices) using an excitation of 485 nm and an emission wavelength of 528 nm. A standard serially diluted FITC-dextran (0, 125, 250, 500, 1,000, 2,000, 4,000, 6,000 and 8,000 ng ml⁻¹) was used for quantification. Fluorescence values were normalized to the control group.

Pontarollo G., Kollar B., Mann A., Khuu M.P., Kiouptsi K., Bayer F., Brandão I., Zinina V.V., Hahlbrock J., Malinarich F., Mimmler M., Bhushan S., Marini F., Ruf W., Belheouane M., Baines J.F., Endres K., Reba S.M., Raker V.K., Deppermann C., Welsch C., Bosmann M., Soshnikova N., Chassaing B., Bergentall M., Sommer F., Bäckhed F, & Reinhardt C. (2023). Commensal bacteria weaken the intestinal barrier by suppressing epithelial neuropilin-1 and Hedgehog signaling. Nature Metabolism, 5(7), 1174-1187.

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