Multisizer II Reference Manual (1998)

Section 7 Additional Techniques

7.1 SMALL APERTURE TECHNIQUES (5Oµm or less)

The comments and cautions in this section are to be used as a procedural

guide to the use of small orifice tubes only, i.e. those less than 5Oµm in
diameter.

It is inadvisable to use the automatic current and gain setup (Analysis Setup
-1 menu) as the aperture current selection process may result in damage to
small apertures. It will be necessary to use the manual current and gain
setup. Typically, 800µA should be regarded as the maximum working
current. Though there will be circumstances where 1000µA on a 3Oµm
aperture and 1600µA on a 5Oµm aperture will be acceptable. The aperture
current should never exceed 800µA with apertures smaller than 3Oµm.

The main point to remember about small apertures is that the background
count should be as low as possible, since the instrument is working near the
limits of its capabilities. There are several sources of high background
counts indicated below 50µm.

7.1 .1 SOURCES OF BACKGROUND NOISE

7.1 .1 .1 Dirty Electrolyte Solution

The factor which influences the successful use of small apertures more than
any other is the strength and cleanliness of the electrolyte solution.

The choice of electrolyte solution will depend upon the nature of the material
to be sized. Scientific instrument laboratory specialists at Coulter Electronics
Ltd will be pleased to make suggestions. For most electrolyte solutions, it
will be found that increasing the salt concentration will increase signal to
noise ratio at sizes below 1 µm. .

For example: 2-4% NaCI instead of 1 % NaCI

4 or 6% Na3 P04 instead of 2% Na3 PO4.

It should be noted; however, that an increase in electrolyte solution

concentration may have an adverse effect upon the material being
analyzed, and at all stages this should be checked.

When deciding upon the concentration of the electrolyte solution, it should

be remembered that one of the limiting factors is the aperture resistance.
 Ideally the aperture resistance of all orifice tubes should be in the range 1k-
100k ohms.

Filtration of the electrolyte solution is of extreme importance and the choice

of filtration equipment available for this type of work is fairly limited (see
Appendix 3). The procedure in general use is detailed in the ‘Fine Particle
Application Notes’ for COULTER COUNTER® Instruments Section 1, Para.
1.4.

7.1.1.2 Atmospheric Interference

It will be virtually impossible to obtain any satisfactory counts if the

instrument is located in a dust-laden atmosphere. The instrument should
preferably be kept in either a reasonably dust-free room or enclosed in a
clean air cabinet.

7.1.1.3 Electrical Interference

An increased background count will be caused by any form of electrical

interference. The most obvious symptom of electrical interference is random
noise patterns and pulses occurring on the Scope Display. Also the
Concentration Index will tend to increase its reading and may also be
unsteady when the instrument is subject to electrical interference. Any
deviation from the normal should be immediately obvious. Mains-borne
interference may be eliminated by moving the instrument to a different
power supply or by installing a mains filter.

7.1.1.4 Stirrer Motor

Another source of noise can be the stirrer motor. When using the instrument
at 2% of the aperture diameter it may help to turn the motor off whilst
performing the analysis. (Small particles will not settle out in this short time.)

7.1.1.5 Damaged Aperture Wafer .

A further possible source of noise is the orifice tube itself; this will normally
only show at the smallest size levels as variable noise on a blank electrolyte
solution, where none had been present before. If the orifice tube is the
cause, then the blank count will tend to differ considerably from one aperture
current polarity to the other. Fit another orifice tube with the same aperture
diameter into the stand, and repeat the test.

7.1.1.6 Excessive Aperture Current

Excessive current causes the electrolyte solution in the aperture to heat, or

even to boil, causing noise. This can normally be seen on the aperture
 viewing monitor as either a dark plume of fluid or as bubbles leaving the
aperture. The cure is to manually reduce the current or to increase the
conductivity of the electrolyte solution. Prolonged boiling will permanently
damage the aperture wafer (see paragraph 7.2.5).

7.1 .1 .7 External Sound and Mechanical Vibration

With the higher aperture resistances that are encountered with small
apertures, the instrument can be affected by either vibration of the
supporting bench or by local sounds of high intensity (banging, shouting,
etc) and these should be eliminated during the analysis. This vibration can
be sensed and magnified by bubbles clinging to the inside of the orifice tube
and the aperture wafer. Any bubbles should be flushed away by opening
both controls on the Sampling Stand as follows:

(1) Set RESET/COUNT to RESET.

(2) Momentarily set FILL/CLOSE to FILL.

The use of a suitable surfactant solution inside the orifice tube will also
alleviate the problem.

7.1 .2 SPECIAL PROCEDURES WHEN USING APERTURES SMALLER THAN

3Opm

When measuring particles with a diameter approaching the lower limit of the
instrument (approximately .4 µm), all factors regarding the elimination of
noise sources assume an even greater importance, and other factors are
introduced.

It is therefore recommended that the procedure adopted to measure these

small diameters takes into consideration the following points:

(a) The electrolyte solution should be membrane filtered twice at .2µm and,
where possible, should consist of a 4% saline solution or one of
equivalent conductivity.

(b) Ensure that the aperture current never exceeds 800µA. High currents
can erode the aperture as well as generate electrical noise.

(c) Always position the Sampling Stand to the right of the electrode module.
Computer monitors should be positioned as far as is practically possible
from the Sampling Stand.

(d) When the mercury has been unbalanced, prior to each count sequence,
 set RESET/COUNT to COUNT. This will remove all pulsations from the
mercury in the manometer as it re-balances and thus eliminate a possible
source of electrical and mechanical noise. If MANOMETER SELECT is set to
2000µl, several timed counts may be made before the aperture current is cut
off when the mercury reaches this contact.

7.2 LARGE APERTURE TECHNIQUES

When using aperture tubes with aperture diameters 560µm, 1000µm or

2000µm the addition of a certain percentage of thickening agent, for
example glycerol, to the electrolyte solution is necessary to reduce noise
produced by turbulent flow of the electrolyte solution through the aperture.
The 400µm aperture design and configuration produce a ‘border line case’
i.e., some aperture tubes require the addition of, for example, 1 0% glycerol
to the electrolyte solution to prevent noise while others function perfectly well
with low viscosity electrolyte solutions, for example lSOTON II diluent. It
must be emphasized that nothing is physically wrong with a new 400µm
aperture tube which requires an electrolyte solution of increased viscosity. In
fact the addition of glycerol has a two-fold effect; by increasing viscosity the
turbulent flow is reduced and at the same time the suspending of large,
dense, particles is facilitated.

Note: The size levels obtainable with 1000µm and 2000µm aperture tubes
are very dependent upon the density of the material being analyzed
and the practical upper limit with the COULTER® Multisizer II is in
the order of 1200µm with particles of density approximately I g/ml.

(1 ) Modified handling of the vacuum system and special precautions for

particle suspension may be required.

(2) Figure 7-1 shows one method of supplying a constant vacuum source.
The tube from the large vacuum reservoir should have a pinch-off clamp
so that the vacuum storage can be preserved whenever the system is to
be open to atmosphere for any reason. It will be found that the mercury
cannot be drawn back beyond the ‘start’ contact; to overcome this, the
following procedure must be observed:

(a) Set the Sampling Stand MANOMETER SELECT control to OUT, to

isolate the manometer contacts. The external electrode will remain
live.

(b) Press the RESET button on the Multisizer II, set RESET/COUNT on
the Sampling Stand to RESET and then press the START switch.
 Alternatively, use a larger pump and fit a vacuum gauge directly on
to the reservoir to monitor the vacuum.

(3) Large sample beakers are helpful. Extra sample suspension can be
added from an agitated sample supply container.

(4) The beaker platform may be lowered and a large sample beaker placed
on it.

(5) Turbulence due to fast flow of low viscosity electrolyte solutions through
the 2000, 1000, 560 and sometimes 400j.tm orifices can create an
audible noise which can actually be heard as a ‘whistle’ and can be
picked up as ‘noise’ on the Multisizer II. To overcome this, electrolyte
solutions can be used with the addition of, for example, 30% glycerol for
the 2000 and 1000µm orifices and 10% glycerol for the 560 and 400µm
orifices when analyzing large, low density particles, e.g. PVC.

(6) Large dense particles are difficult to suspend, and normal vacuum levels
(150 to 160 mm of mercury) may cause turbulence that can interfere with
the analysis. Increasing the viscosity or density of the electrolyte solution
substantially overcomes this problem as does reducing the vacuum
level. Viscosity should not generally be higher than the equivalent of
90% glycerol, nor should the vacuum applied to the aperture be reduced
to less than 70mm of mercury.