Zetasizer Nano-ZS User Instructions

1. Activate the instrument computer by logging in to the central login system in the
service corridor and then log into local instrument computer Username: zetasizer
Password zetasizer

2. Instrument is usually on. If the instrument is off, turn on the instrument and wait 30
minutes for the laser to stabilize.

a. Unstable light source is more likely to introduce errors in the measurements.

b. When turning on the instrument, a beep will occur to indicate the instrument
has been turned on, followed by another beep has finished the setup routine.

c. Two further beeps will be heard to indicate the instrument has reached the
default temperature of 25C.

3. Start the Zetasizer software by clicking the DTS desktop icon:

4. If measurement data from the previous user shows up it is important that you remove
this

a. From the File menu select close until all the measurement data is removed from
the window
OR
b. Click the lower X in the right corner of the data window until all your
measurement data is removed from the window

5. Choose the appropriate cuvette cell for your sample and measurement type.

a. For Particle Size Measurements use DTS0112 Low volume cuvette

b. For Zeta Potential Measurements use DTS1060C Clear Zeta Cell

c. Guides on cuvette use are in manual - page 4.3

d. The plastic cuvettes should not be used above 50
o
C

e. The zetapotential plastic cell should not be used above 70
o
C

f. Both cuvettes are only compatible with H
2
O and/or Ethanol solutions


6. Fill your selected cuvette with your prepared sample


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a. DTS0112 Low volume cuvette for size measurement needs ~375L. Check that
all air bubbles are removed.

b. DTS1060C Clear Zeta Cell should be slowly filled by syringe with ~0.75mL of
sample. Check that all air bubbles are removed.

7. Open the lid to the zetasizer by pressing the metallic button


8. Insert cuvette in instrument

a. If the cuvette has an arrow on it, the arrow should be faced against you. Be sure to
check the solution level against the sticker on the inside of the lid.

b. Sample preparation is key to obtaining good data. Information on sample
preparation can be found in manual chapter 6

!!! CAUTION NEXT STEP IS IMPORTANT FOR CORRECT OPERATION !!!

9. Confirm that there is no measurement data from the previous user displayed in the
window.

10. If there is, oo to File menu and select Close to remove measurement data from previous
user

11. Create a personal folder if needed in C:\Users\netid or C:\Program Files\Malvern
Instrument\DTS\Measurement Data\netid

12. Create new measurement file #.dts

13. Go to Measure menu and select Start SOP or Manual

14. Select each parameter tab ( click on Advice button, or see Advice Notes below)

a. Measurement type
b. Labels
c. Cell
d. Sample
e. Temperature
f. Measurement

15. Choose the appropriate cell(s) to match the one youre using.

For Particle Size Measurements Use DTS0112 Low volume cuvette





For Zeta Potential Measurements Use DTS1060C Clear Zeta Cell





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16. Save as SOP.. if you want to save these settings

17. Click Ok

18. Measurement window will now show on screen


19. Make a manual measurement by pressing Start.

20. Check the Count Rate and the Correlation factor as measurements are being taken. See
manual pages 4.20-4.22 for advice on interpretation.

21. When measurements are completed the system will beep.

22. Review your measurements to ensure you have all data.

!!! CAUTION NEXT STEP IS IMPORTANT FOR CORRECT OPERATION !!!

23. Unload measurement data from the DTS software before you close down

a. From the File menu select close until all your measurement data is removed from
the window
OR
b. Click the lower X in the right corner of the data window until all your
measurement data is removed from the window

24. Remove cuvette

25. Clean the cuvette(s) you have used see below for more details

26. When done, close the software, put all parts back in drawer, clean up benchspace and
log out of local computer +central log system.


Cleaning the cuvettes

When done, be sure to clean the cuvettes properly for the next user.

1. Disposable plastic cuvettes
a. Discard solvent appropriately and dispose of cuvettes


2. Zetapotential plastic cuvette

a. The cuvette is semi-disposable and is usually good for ~40 measurements or until
the electrodes are no longer functional.

b. To clean the cuvette, use a luer lock syringe to rinse with 2x10 ml of the same
solvent used for the measurement followed by 3x10 ml 18.2 Mohm water. Dry
with nitrogen

3. Universal dip cell

a. Only use this cuvette prior to talking to staff. Special instructions required.


4. Glass/quarts cuvettes

a. The cleaning procedure for glass or quartz cuvettes is dependent on the sample
that was measured, so cleaning is sample specific. However, the following advice
should be followed.

b. Rinse the cuvette with the same dispersant that was used for the measurement, i.e.
if the sample was dispersed in water - use clean water to rinse it.
c. Clean the cuvette in an ultrasonic bath of clean solvent. rinse with 5-10 volumes
of 18.2M ohm DI water.

d. Once clean, dry the cuvette with nitrogen.

e. The smaller and more dilute the sample being measured, the more important the
cleanliness of the cuvette.

f. Be careful not to scratch the surface of ANY cuvette!


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Material properties - Advice
The sample refractive index and absorption are only used in the conversion of the intensity
distribution to a volume or number distribution. They have no effect on the calculation of the Z-
Average mean and polydispersity.
The accuracy of the refractive index of the material is only required to +/- 0.05 units if within 0.2
units of the refractive index of the dispersant. For materials with a difference in RI values greater
than 0.2 units from the dispersant, an accuracy of 0.1 is sufficient.
Dispersant properties - Advice
The choice of dispersant is usually dictated by the medium the sample is already dispersed in. If
the sample has to be diluted for measurement, the original dispersant should be imitated as
closely as possible. Preferably the dispersant should be extracted from the sample and a portion
of the sample redispersed in it.
A dispersant can be added using the dispersants button. As the parameters for any added
dispersant will only be applicable at a specific temperature, the temperature selected will be
displayed in the dispersant/solvent properties box.
The technique relies on the measurement of the Brownian motion of the particles or molecules in
the dispersant. This is reduced at higher viscosity, so the results will be most repeatable with
dispersants of low viscosity such as water, and much less repeatable with viscosities above
100cP. This rule of thumb is highly size dependant. Particles less than 10nm have been
successfully measured at over 1000cP, whereas particles over 1 micron will need to be in low
viscosity dispersants to be measured at all.
The size result will be wrong if an incorrect dispersant refractive index is used.
In general, a refractive index error of less than 0.005 will affect the size accuracy by less than
1%.
The size reported is directly proportional to the viscosity. A viscosity used that is 50% higher
than the actual sample viscosity, will report a size that is 50% too low.
Temperature settings - Advice
Temperature
To speed up the equilibration of sample temperature, it is helpful to set the temperature to that of
the laboratory, unless there is a specific reason to measure at another temperature.
The size of most samples will be unaffected by the setting of the temperature over a range of a
few degrees, as long as the viscosity has been corrected appropriately.
This will not be the case for samples such as micelles, proteins and polymers, where the
conformation, and therefore the size, may be highly temperature dependent.
If it is unknown if the sample has a size dependence on temperature, it is good practice always to
measure at the same temperature to ensure comparability. Alternatively, conduct test
measurements at a range of temperatures.


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Note: As the pH probe is not temperature compensated, it is recommended that titrations are
performed with ambient temperature conditions. Adjust the temperature in the SOP to the
ambient value.
Equilibration time
Entry of an equilibration time adds a delay before the start of a measurement sequence to ensure
the sample temperature is equal to the cell block temperature before the measurement is started.
This delay starts as soon as the system has reached the temperature requested. Allowing the
sample to be fully temperature equilibrated ensures the actual Brownian motion of the particles is
measured and not convection due to thermal gradients. For any temperature change it takes four
minutes for the sample to become stable to within 0.1deg of the requested temperature after the
system indicates that the target temperature has been reached, so this should be considered a
starting point for the most accurate measurements
If you find that the first result of a series of repeat measurements is different to all of the repeated
measurements, try increasing the equilibration time.
When should the cuvette be placed into the instrument?
Selection of After the set temperature has been reached enables the sample to be kept at a
constant temperature, in a water bath for example, and transferred to the system after the system
temperature has equilibrated. The measurement sequence will stop and the status bar will request
the user to insert the sample at the appropriate time.
Measurement duration - Advice
The result quality is linked to the measurement duration. The automatic setting will give a
suitable measurement duration for a repeatable size distribution to be determined for the majority
of samples.
If greater repeatability is required, the manual setting will allow a higher number of runs, and a
longer run duration, which will result in better data quality.
If only the Z-Average size is required, the manual setting can be used to reduce the measurement
time. As with all measurements, a number of repeats are recommended to ensure that the result
repeatability is still acceptable.
The balance between the number of runs and the run duration will be sample dependant. For dust
free samples less than 50nm a run duration of 5 seconds should be sufficient, for larger materials
20 seconds is advised.
An hour, or 180 x 20-second measurements would be considered the longest practicable
duration, except for the study of very slow processes such as those observed in gel systems.
A minimum of 1 run of 1 second is allowed. The maximum is 36 runs of 600 seconds duration
Number of measurements - Advice
It is wise to do multiple measurements on a sample for two reasons: Firstly it will indicate if the
first measurement is different to the rest because the sample temperature had not equilibrated.
Secondly, it is difficult to exclude all the effects of contaminating dust. Even with the dust


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rejection algorithm employed, and measuring at higher concentrations, particles of dust scatter so
much light that if dust happens to be present in all the sub-runs of a measurement, the result may
be substantially affected. This can be spotted as a rogue result in a series of measurements.
If multiple measurements are selected, the same measurement strategy will be used for all
measurements.
A delay between measurements is useful if a size change is being observed over a long period,
e.g. 24 hours, but where a large number of results would be unwieldy and unnecessary.
Append measurement number to sample name adds a sequential number to each sample name,
starting with one. This counter is reset every time a measurement sequence is started.