Three million cells grown on 100-mm tissues culture dishes were detached by 0.25% trypsin-ethylenediaminetetraacetic acid (trypsin-EDTA) and the cell pellets were stored at −80 °C. For an assay, 70 microliters of phosphate buffer (50 mM, pH 7.0) were added to the cell pellet and sonicated for 30 s using a Vibra-Cell sonicator (Sonics, Newton, CT). After sonication, to break up cell membranes, samples were centrifuged for 5 min at 10,000 g in an Eppendorf Microcentrifuge. Five hundred microliters of phosphate buffer (50mM, pH 7.0) were added to a 10 mm pathlength quartz cuvette (Starna Cells, Atascadero, CA, USA), followed by 20 µL of cell lysate and 250 µL of 30 mM H2O2. Catalase activity were measured in a HP UV–Vis Spectrophotometer. The absorbance at 240 nm was followed for 120 s with a cycle time of 10 s. The activity of catalase in cells was determined as mkUnits per million cells [20 (link),21 (link)].
10 mm pathlength quartz cuvette
Manufactured by Starna Cells
Sourced in United States
The 10 mm pathlength quartz cuvette is a laboratory equipment designed for spectroscopic analysis. It is made of high-quality quartz material and provides a consistent 10 mm optical path length for samples. The cuvette is commonly used in various analytical techniques, such as UV-Vis spectroscopy, to measure the absorbance or transmittance of solutions.
Automatically generated - may contain errors
2 protocols using 10 mm pathlength quartz cuvette
1
Catalase Activity Assay in Cell Lysates
2
Upconverting Nanoparticle pH Cycling
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A CW
980 nm diode laser (Opto Engine) is fiber-coupled and focused onto
a 10 mm path length quartz cuvette (Starna Cells, Inc.) through a
collimator with an N-BK7 Plano convex lens (f = 20.0
mm) and an additional Plano convex lens (f = 35.0
mm) from Thorlabs. The incident irradiance is estimated to be 100
W/cm2 with a power of 800 mW and beam diameter of 1 mm.
Emission is collected after a 750 nm SP filter by an OceanOptics HR4000
spectrometer.
For pH cycling, 0.48 M HCl and 0.48 M NaOH solutions
in DI water are prepared. pH is measured for a test sample containing
S-Medium buffer to calibrate the volume of acid or base necessary
to tune pH across the relevant range (pH 3 to pH 6) for five cycles.
On the basis of the calibration, HCl and NaOH are added dropwise to
the cuvette containing 1 mL of UCNPs in S-Medium (10 mg/mL). Spectra
is collected after shaking the cuvette using the setup mentioned above.
For intensity corrections, x is added to the raw
normalized intensity values. . This calculation assumes that intensity
and concentration are linearly related.
980 nm diode laser (Opto Engine) is fiber-coupled and focused onto
a 10 mm path length quartz cuvette (Starna Cells, Inc.) through a
collimator with an N-BK7 Plano convex lens (f = 20.0
mm) and an additional Plano convex lens (f = 35.0
mm) from Thorlabs. The incident irradiance is estimated to be 100
W/cm2 with a power of 800 mW and beam diameter of 1 mm.
Emission is collected after a 750 nm SP filter by an OceanOptics HR4000
spectrometer.
For pH cycling, 0.48 M HCl and 0.48 M NaOH solutions
in DI water are prepared. pH is measured for a test sample containing
S-Medium buffer to calibrate the volume of acid or base necessary
to tune pH across the relevant range (pH 3 to pH 6) for five cycles.
On the basis of the calibration, HCl and NaOH are added dropwise to
the cuvette containing 1 mL of UCNPs in S-Medium (10 mg/mL). Spectra
is collected after shaking the cuvette using the setup mentioned above.
For intensity corrections, x is added to the raw
normalized intensity values. . This calculation assumes that intensity
and concentration are linearly related.
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