Two New Options for Measuring Small Particles
Measurements of weakly scattering small-volume samples is now much easier, more convenient, and less expensive with the 90Plus or the BI-MAS option on the ZetaPlus or the ZetaPALS . Using the optional BI-APD , avalanche photodiode detector, and the BI-SM50 , small volume cell, particles with hydrodynamic radii as small as 1.5 nm are now measurable.
The BI-APD has more than ten times the sensitivity of the standard PMT detector at the wavelength of the standard 30 mW laser. Thus, the count rate with the BI-APD is equivalent to a laser with more than 300 mW power. With such sensitivity, measurements on very weakly scattering particles are possible without the need for a more costly laser.
That is only half the story, especially with precious samples like proteins. Often only a milligram or two of sample is available for light scattering studies. If diluted into the normal 2 mL of liquid, the concentration is then too low for good measurements. The answer is the 50 µL BI-SM50 disposable cell . This cell fits directly into the 90Plus on top of a custom adapter, the BI-SM50A . Using the insertion/removal tool supplied with each one, the adaptor is easily removed to allow use of the standard cells.
A couple of drops constitute 50 µL. The open-well design allows easy removal of the samples after the measurement for sample reconstitution, if desired. The sample is not diluted as it would be with a flow cell.
Measurements with Lysozyme*
Lysozyme is a small, globular protein, well characterized and widely available. Measurements were done at 13.6 kDa, pH 6, in 10 mg/mL NaCl, at 4.6, 9.2 & 15 mg/mL of Lysozyme. The mean hydrodynamic radius, R h , 1.77 ± 0.12 nm. Using the crystal structure, assuming a prolate ellipsoid, this result agrees well with one layer of water attached to the lysozyme surface.
Without the BI-APD , the photon count rate is quite low from such a molecule. The resulting noisy autocorrelation function does not yield reproducible answers with a standard laser. With the BI-APD , excellent repeatability in the size is obtained.
The measurement shown below was made with 9.2 mg/mL in the BI-SM50 cell. The amount of protein used is therefore less than 0.5 mg. And, it is recoverable .

Summary Report of Measurement of Lysozyme in Small Volume Cell
with BI-APD High Sensitivity Detector

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