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Hts 7000 plate reader

Manufactured by PerkinElmer
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The HTS 7000 plate reader is a versatile and high-performance instrument designed for a wide range of microplate-based assays. It features multi-mode detection capabilities, including absorbance, fluorescence, and luminescence, allowing researchers to conduct various assay types. The HTS 7000 provides accurate and reliable data acquisition, enabling efficient and precise measurement of samples across multiple microplate formats.

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Lab products found in correlation

Calcein am, by Thermo Fisher Scientific (2 mentions) Fluoroblok insert, by BD (1 mentions) 2 nbd glucose, by Thermo Fisher Scientific (1 mentions) Insulin, by Merck Group (1 mentions)

4 protocols using hts 7000 plate reader

1

Neutrophil Chemotaxis Assay Using FluoroBlok

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Chemotaxis of purified neutrophils was assessed by means of FluoroBlok inserts (Falcon; BD). Cells (5 × 106/ml) were labeled with calcein-AM (1 µM final concentration; Molecular Probes) for 30 min at 37°C, washed twice, and resuspended in Hepes buffer at a concentration of 2 × 106/ml. Chemoattractant solution (PAF, IL-8, and C5a, all at 10 nM; Sigma-Aldrich) or medium alone (0.8 ml/well) was placed in a 24-well plate, and 0.3-ml cell suspension was delivered to the inserts (3-µm pore size) and placed in the 24-well plate. Cell migration was assessed by measuring fluorescence in the lower compartment at 2.5-min intervals for 45 min with the HTS7000+ plate reader (Perkin Elmer) at an excitation wavelength of 485 nm and emission wavelength of 535 nm. Maximal slope of migration was estimated over a 10-min interval.
Canault M., Ghalloussi D., Grosdidier C., Guinier M., Perret C., Chelghoum N., Germain M., Raslova H., Peiretti F., Morange P.E., Saut N., Pillois X., Nurden A.T., Cambien F., Pierres A., van den Berg T.K., Kuijpers T.W., Alessi M.C, & Tregouet D.A. (2014). Human CalDAG-GEFI gene (RASGRP2) mutation affects platelet function and causes severe bleeding. The Journal of Experimental Medicine, 211(7), 1349-1362.
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2

Microencapsulated Protein Release Kinetics

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AlexaFluor488-labeled IgG (goat anti-rabbit IgG, Life Technologies), bovine serum albumin-AlexaFluor488 conjugate (Life Technologies), 2-NBD-glucose (Life Technologies) or insulin (Sigma) tagged with AlexaFluor488 was dissolved in a 5% PEG-4MAL (10 kDa or 20 kDa) solution before being microencapsulated by macromer droplet gelation. To prevent proteins from being crosslinked by the macromer, thiols were capped using aminoethylate reagent (Thermo Scientific) according to product instructions. Particles were washed and resuspended in PBS and divided into 5 replicates containing 2 mL total volume. 50 µL samples were taken of supernatant alone, as well as of supernatant containing well-mixed, protein-laden microgels. These samples were placed in a 96 well plate, and their fluorescent intensity was measured using a Perkin Elmer HTS 7000 plate reader. To generate release curves, supernatant samples were collected over the course of 3 days, and their fluorescent intensity was measured. Protein release was normalized by setting fluorescent intensity of the supernatant alone correspond to 0% protein released, and fluorescent intensity of the buffer/microgel mixture correspond to 100% protein released. This data was plotted using GraphPad Prism, and exponential best fit curves were calculated from normalized data.
Headen D.M., Aubry G., Lu H, & García A.J. (2014). Microfluidic-Based Generation of Size-Controlled, Biofunctionalized Synthetic Polymer Microgels for Cell Encapsulation. Advanced materials (Deerfield Beach, Fla.), 26(19), 3003-3008.
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3

Chemotaxis Assay for BMDM and BMDN

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Chemotaxis of BMDM and BMDN was assessed by means of Fluoroblock inserts (Falcon, Colorado Springs, CO, USA). 6*105 BMDN or BMDM from WT and SIRPαΔcyt mice were labeled with calcein-AM (Molecular Probes, Invitrogen, Carlsbad, CA, USA) and seeded on the upper chamber of 3μm or 8μm pores respectively. C5a (10nM) was used as a chemoattractant. Cell migration was assessed by measuring fluorescence in the lower compartment at 2’ intervals for 1h (BMDN) or 2h (BMDM) with a HTS7000+plate reader (Perkin Elmer, Waltham, MA, USA) at an excitation wavelength of 485 nm and emission wavelength of 535 nm.
Alvarez-Zarate J., Matlung H.L., Matozaki T., Kuijpers T.W., Maridonneau-Parini I, & van den Berg T.K. (2015). Regulation of Phagocyte Migration by Signal Regulatory Protein-Alpha Signaling. PLoS ONE, 10(6), e0127178.
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4

HDL Antioxidant Capacity Measurement

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HDL was isolated from EDTA plasma by ultracentrifugation for measurement of triglyceride, cholesterol and protein content. HDL triglyceride and HDL cholesterol content was expressed as nmol per gram protein (nmol/g). HDL‐TG was calculated as the HDL triglyceride: total cholesterol ratio and expressed as nmol/nmol. The anti‐oxidative function of HDL was assessed by monitoring its capacity to inhibit the oxidation of dichlorodihydrofluorescein (Invitrogen) by oxidized 1‐palmitoyl‐2‐arachidonoyl‐phosphathidylcholine (Avanti Lipids).9 Briefly, in a total volume of 790 µL phosphate‐buffered saline (pH 7.4), 35 µL of a normal LDL solution (final concentration of 18 µmol/L cholesterol), 35 µL of test HDL cholesterol (final concentration of 11 µmol/L cholesterol), 20 µL oxidized 1‐palmitoyl‐2‐arachidonoyl‐sn‐phosphatidylcholine (final concentration 50 µmol/L), and 10 µL of dichlorofluorescein solution (final concentration of 40 µmol/L) were incubated in glass tubes for 2 hours at 37°C. Afterwards the fluorescence intensity was determined with a HTS 7000 plate reader (Perkin Elmer) at 485 and 530 nm (excitation and emission wavelength respectively). The intra‐assay CV was 3.4%.
Verwer B.J., Scheffer P.G., Vermue R.P., Pouwels P.J., Diamant M, & Tushuizen M.E. (2020). NAFLD is related to Post‐prandial Triglyceride‐enrichment of HDL Particles in Association with Endothelial and HDL Dysfunction. Liver International, 40(10), 2439-2444.
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