For the measurement of chlorophylls and total carotenoid content, Stevia leaves were harvested from the fourth and fifth nodes of plants that were grown in the greenhouse for 3 weeks and frozen in liquid nitrogen. After homogenization, 200 mg of the powdered leaves was extracted twice with 2 mL of 100% methanol at room temperature for 1 h with constant shaking in the dark. The extracts were pooled and diluted fivefolds before analysis on an Infinite M2000 microplate reader (Tecan, Männedorf, Switzerland). Absorbance values at three different wavelengths, 666 nm, 653 nm and 470 nm, were used to calculate the relative amount of chlorophyll a, chlorophyll b and total carotenoids present in the leaves based on previously reported formula (Lichtenthaler and Wellburn, 1983 ).
Infinite m2000 microplate reader
Manufactured by Tecan
Sourced in Switzerland
The Infinite M2000 microplate reader is a versatile laboratory instrument designed for measuring various types of samples in microplate format. It is capable of performing absorbance, fluorescence, and luminescence measurements, providing a comprehensive solution for a wide range of applications in life science research and clinical diagnostics.
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Lab products found in correlation
3 protocols using infinite m2000 microplate reader
1
Measurement of Leaf Pigments in Stevia
2
Quantifying Ligand Density on PEG-Alkyne Surfaces
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The ligand density on PEG-alkyne-modified
surfaces is quantified by adopting a method established by Barber
et. al.32 (link) Briefly, glass substrates coated
with varying mol % of PEG-alkyne are modified by the click reaction
with a fluorescently labeled peptide, K(N3)GGNGEPRGDTYRAYK(fluorescein)GG,
which contains a chymotrypsin digestion site. The coating on the surfaces
is enzymatically digested with α-chymotrypsin by treating the
substrate with 1.25 μg/mL α-chymotrypsin in 30 mM Tris
HCl (pH 8.00), 50 mM CaCl2, and 10 μM HCl for 16
h at 30 °C. The surfaces that have been enzymatically digested
and the untreated samples are mounted with Mowiol, and the fluorescence
intensity on the surfaces is measured. The fluorescence intensity
of the solution above the digested surface is measured with a microplate
reader (Infinite M2000 microplate reader, TECAN, Männedorf,
Switzerland), and the peptide concentration in solution is calculated
by using solutions of the fluorescently labeled peptide with known
concentrations
surfaces is quantified by adopting a method established by Barber
et. al.32 (link) Briefly, glass substrates coated
with varying mol % of PEG-alkyne are modified by the click reaction
with a fluorescently labeled peptide, K(N3)GGNGEPRGDTYRAYK(fluorescein)GG,
which contains a chymotrypsin digestion site. The coating on the surfaces
is enzymatically digested with α-chymotrypsin by treating the
substrate with 1.25 μg/mL α-chymotrypsin in 30 mM Tris
HCl (pH 8.00), 50 mM CaCl2, and 10 μM HCl for 16
h at 30 °C. The surfaces that have been enzymatically digested
and the untreated samples are mounted with Mowiol, and the fluorescence
intensity on the surfaces is measured. The fluorescence intensity
of the solution above the digested surface is measured with a microplate
reader (Infinite M2000 microplate reader, TECAN, Männedorf,
Switzerland), and the peptide concentration in solution is calculated
by using solutions of the fluorescently labeled peptide with known
concentrations
3
Quantifying hBM-MSCs Adhesion and Proliferation on 3D Scaffolds
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Immunofluorescence staining was performed to study the adhesion and proliferation of hBM-MSCs cells on the scaffolds. The cell-seeded scaffolds were washed with PBS and fixed with 3.7% (v/v) formaldehyde for 10 min after 7 days of culture. The cells were permeabilized with 0.1% (v/v) Triton X-100 for 5 min and the blocking of the samples was performed with 1% (w/v) BSA for 30 min. The F-actin filaments were stained with 165 nM AlexaFluor 488-labelled phalloidin for 30 min at room temperature protected from light. After washing the samples with PBS three times, the nuclei were stained with 300 nM DAPI solution. The samples were observed under inverted fluorescence microscope (Nikon TMS).
The proliferation of hBM-MSCs on the 3D scaffolds was evaluated by means of metabolic activity determination following CCK-8 assay. The cell-seeded samples were rinsed with PBS after 2, 7 and 10 days of incubation and 350 μL of fresh medium containing 35 μL of CCK-8 kit reagent was added to each scaffold. The samples were incubated for 4 h at 37 °C and the optical density of the generated formazan was measured by Tecan Infinite M2000 microplate reader.
The proliferation of hBM-MSCs on the 3D scaffolds was evaluated by means of metabolic activity determination following CCK-8 assay. The cell-seeded samples were rinsed with PBS after 2, 7 and 10 days of incubation and 350 μL of fresh medium containing 35 μL of CCK-8 kit reagent was added to each scaffold. The samples were incubated for 4 h at 37 °C and the optical density of the generated formazan was measured by Tecan Infinite M2000 microplate reader.
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