Count and size more cells for smoother averaged size distribution and greater statistical confidence in results. Contact Us. Sign in to view contract pricing Your shopping cart is empty. Qty Price View Cart. Added to cart View Cart. Note: All results are automatically corrected for coincidence loss. The result allows the analyses to be expressed in terms of the original sample concentration.
Dilution factor A numeric field for entering the factor conversion of count results to. Operation concentration. If a value of 0 is entered or the field is left empty, the factor defaults to 1.
A dilution factor of up to four digits including one decimal point can be entered directly. For a 50, dilution, the figures entered in the dilution factor line in screen A1 should be: Dilution factor: 5. The units remain switched throughout the Z Series menus, until switched back.
A selection field giving you the option of selecting from three channelyzer resolutions: 64 channels, channels or channels. The default is channels. All printed and displayed graphs and tables use the selected resolution. The graph displays the full width of the accumulation, channels. If you selected or 64, the graph still displays the full width of the accumulation but combines sums adjacent channels. For example, the first displayed column contains the sum of the first two if you selected or four if you selected 64 channelyzer channels.
The second displayed column contains the sum of the third and fourth if you selected or fifth through eighth if you selected 64 channelyzer channels. If you select Automatic, at the end of the analysis the results are automatically printed. If you select Short, at the start of the first analysis a columnized printout with the instrument settings as headings is produced.
Instrument settings are not printed again until one or more are changed. If you select Long, a printout of all the instrument settings with each individual result is produced. Graph The available selections for this field are controlled by the printer that you select and the value selected in Form.
Refer to Table for valid selections. Pressing PRINT sends a copy of the current result to the printer whether in manual or automatic mode. Results can be printed in one of two formats, Short or Long.
Operation Table The available selections for this field are controlled by the printer that you select and the value selected in Form. Results can be printed in one of two formats Short or Long. Various status messages appear in the Message area of the Data Terminal.
Table 4. Tl represents the lower size setting and Tu the upper. For optimum analysis results, the concentration should be as low as practicable e. Shows the heights of a random selection of pulses on a scale of 0 to units, along with an indication of baseline noise levels, and horizontal line s to show the position of the Lower Size Tl and the Upper Size Tu Dual Threshold version only.
The number of pulses shown on the display represents the sample concentration. Once START is pressed and screen A2 of the Analysis screens displays, the analysis continues until the selected volume of sample has been analyzed. If Concentration was selected, screen A3 displays: A With the Dual Threshold version, a selection field on both these screens allows access to the three results, above Tl lower size , above Tu upper size , or between between the two sizes.
If Auto is selected on the Mode line of screen A8, the results are sent to the printer when screen A3 or A4 displays. Note: Dual Threshold only - displayed count mode has no effect on printout.
Changing Size Settings in the Analysis Screen The operator can move the size threshold setting s by using the cursor keys, while counting is taking place. This analysis result is invalid and has to be repeated with the new settings.
The procedure is as follows: 1. The Lower Size Threshold is set to The The threshold can now be adjusted by using or? Toggle between the two thresholds using the cursor keys. The selected threshold will be highlighted. The horizontal line s on the pulse display, which represent the threshold level s , move as the threshold s are adjusted a slight delay between pressing the cursor key and the threshold response is quite normal. The metering system continues to draw the sample through the aperture for as long as possible in order to maintain the pulse display.
It will then reset, leaving the pulse display frozen. Provide an on-screen display of the numeric values contained in the channelyzer expressed as a percentage of the total number of pulses channelyzed. Depending on the mode selected, provide a printed graph or table of the distribution collected by the channelyzer during the most recent analysis.
Channelyzer Results This function provides an on-screen graph of channelyzed data. The resulting graph scales the vertical axis to maximize the data display.
No scaling of the horizontal axis is offered. See para. The maximum channel is plotted as a column extending from the very bottom of the graph to one pixel from the top of the graph excluding borders. Cursors are represented by a column of inverted pixels extending from the bottom border to the top border inclusive.
To the right of the graph, the Z2 displays either a cumulative over and under value or statistics based on the results. When Cum. When Stats is selected, statistics. Operation display. Statistics The Z2 displays statistics based on channelyzer results. The range these results cover are defined by the placement of the cursor. As the user moves the cursor, the values displayed show the relative amount of channelyzer data lying above and below the cursor.
To obtain statistics:. These profiles contain the:? Choose a profile name to store your information under by selecting either Profile A, B, C, D, or E, using the cursor keys. The instrument displays a short description below the selection fields.
If the profile selected is not blank, the aperture and threshold settings are displayed. The instrument compares the current settings to the ones in the selected profile. If they are compatible, the current settings are changed to the ones in the selected profile. Note: Pressing any other key aborts the loading of the profile. For the disinfection procedure and additional maintenance procedures for biological applications refer to Appendix 7.
End of Day In non-biological applications, if the instrument is to be used the next day or after a short interval, place a beaker of clean diluent on the platform ensuring that the Aperture and the electrode are submerged in diluent, then set the Power Switch to O off and disconnect from the Mains Supply.
In biological applications particularly White Blood Cell counting it is good laboratory practice to Prime the Aperture using a non-lytic cleaner e. Ensure that the Aperture and the electrode are submerged in the cleaning agent before setting the Power Switch to O off.
Long-Term Shutdown If the instrument is to be left for a long period of time in excess of 2 weeks or is in dry ambient conditions, it is advisable to put distilled or de-ionized water in the beaker in place of the diluent, so that salt deposits are not formed as a result of evaporation. Disconnect the level sensors from the rear of instrument. Empty the jars and thoroughly wash.
If the Diluent tubing is disconnected at the instrument, instead of from the lid, diluent will siphon out of the jar. Link the Waste and Diluent connectors with tubing see Figure to prevent the ingress of contamination.
Disconnect tube from diluent vessel and insert in suitable waste container. Place empty vial at sampling station. A spare connection is provided on the Waste Jar cap to accept the disconnected tube. The number of cycles to completion is shown in the status message. The total procedure takes approximately ten minutes.
The procedure can be aborted by pressing STOP. This causes the Drain System procedure to stop when the current cycle has completed. Filling the System If the instrument and power supply have not been switched off, since the system has been drained, ignore steps 1 to 2 and start at step 3.
Changing the Aperture Tube This procedure must be observed if an Aperture is to be replaced without draining the entire system. Storing the Aperture Tube 1 For short term storage, aperture tubes should be capped and stored wet e.
Then capped and stored in its original packaging. Fitting and Priming the Aperture Tube If the instrument and Power Supply have not been switched off since an Aperture Tube was removed, ignore steps 1 to 4 and start at step 5.
The Vent function must be selected when an Aperture Tube is being fitted, to prevent the possibility of liquid being sprayed from the aperture. Then secure it by fully rotating in the direction shown.
When in place with the Coulter logo to the front, the aperture is slightly angled to produce an optimum image on the aperture viewing screen. Do not attempt to adjust the angle of the aperture. Follow the instructions on the screen, ensuring that the Diluent Jar is at least a quarter full with clean diluent, the platform is raised and the door is closed. Setting the Date and Time It is important that the results you print have the correct date and time displayed on them.
Upon power-up initialization, the Z2 checks to see if its real time clock RTC has been set. If it has, the process proceeds. If you receive this message, the Z2 will not allow you to continue until date and time have been entered and START has been pressed. Note: The Z2 does not perform automatic daylight savings time adjustments due to the differences in daylight savings worldwide.
INDEX aperture table, suitable metered volumes for selected aperture sizes. Operation Drain System F5. Guaranteed Data The following data is guaranteed when the instrument is operating within the environment now specified: Input Voltage within Set Range. The instrument is placed on a bench that is not subject to: 1 Strong vibrations or sounds of high intensity e. Each threshold i. Lower and Upper is resolvable and selectable in 0.
Note: The Dual Threshold is an optional extra. Specifications During accumulation, displays the incremental count. Printer Interface. The data output can either be results only or results with instrument settings.
Particle Size Range. Particle Count. Aperture Tube Sizes. Metering Volumes. Data Output. Beckman Coulter Method of Counting and Sizing The Beckman Coulter method of counting and sizing is based on the detection and measurement of changes in electrical resistance produced by a particle or cell suspended in a conductive liquid diluent traversing through a small aperture.
When particles or cells are suspended in a conductive liquid, they function as discrete insulators. When a dilute suspension of particles is drawn through a small cylindrical aperture, the passage of each individual particle momentarily modulates the impedance of the electrical path between two submerged electrodes located on each side of the aperture. Figure illustrates the passage of a particle through an aperture.
An electrical pulse, suitable for counting and sizing, results from the passage of each particle through the aperture. The effective resistance between the electrodes is due to the resistance of the conductive liquid within the boundaries of the aperture. Theoretical and practical analysis of the behaviour of particles within an aperture shows that the height of the electrical pulse produced by the particle is the characteristic that most nearly exhibits proportionality to the cell volume Eckhoff , Grover et al.
This method permits the selective counting of particles and cells within very narrow size-distribution ranges by electronic selection of the pulses they generate. Coincidence Correction Occasionally, more than one particle is within the boundaries of an aperture at the same time coincidence. When this occurs, only one larger pulse is generated. This results in low particle count and high particle volume measurements.
However, the frequency of coincidence is a statistically predictable function of particle concentration, and is corrected by the instrument. Effect of Diluent Electrolyte Solution In a counting system highly sensitive to the volume of the individual particles being counted, the conductive liquid in which the particles or cells are suspended must have a minimum influence on their integrity, and, hence, on their size.
It then counts the number of pulses, as a known volume of sample passes through the aperture. The count is then corrected for coincidence errors e. To achieve this purpose, the instrument contains circuitry that, on command from the software, controls the Hydraulics System, automatic calibration facilities, and an output interface to an optional printer. The following description is based on the simplified block diagram in Figure Pulse Generation A constant aperture current, with a value that is either set automatically or chosen from the Setup Menu is generated by the Aperture Current Generator and routed via the safety circuits to the.
Principles of Operation external electrode. The safety circuits are controlled by the Screened Door and Beaker platform micro-switches. These switches are closed only when the Screened Door is closed and the Beaker Platform is fully raised. The safety circuits prevent the external electrode from being touched while current is flowing.
With aperture current flowing between the external and internal electrodes, a known volume of sample is drawn through the aperture. As each particle passes through the aperture, the impedance between the electrodes changes, causing a pulse whose size is essentially proportional to the volume of the particle. This pulse is routed, via the isolation capacitor, to the preamplifier.
The Dual Threshold facility is an optional extra, therefore all text references to the Upper Threshold, Difference Circuit and Difference Counter only apply if that option is fitted. After amplification, the resultant voltage pulses are sent to threshold comparators, which only allow pulses through that are equal to, or exceed, the size setting. This output is then routed to the Difference Counter. The duration of this signal is determined by the selected volume of sample passing through the aperture.
A sudden absence of pulses before the end of a count would give an immediate indication of a blocked aperture.
In addition to the two counters found in the Z1, the Z2 adds the ability to channelyze pulses. This process continues until the metered volume has been drawn through. Principles of Operation the aperture. The resulting histogram shows the distribution of particle sizes. Figure shows the relationship of these circuits to the Hydraulic System in a very simplified manner. Beckman Coulter Method of Counting and Sizing. Coincidence Correction. Effect of Diluent Electrolyte Solution.
Pulse Generation. Pulse Processing. Control Circuits. INDEX basic principle. Coulter Method of counting and sizing. Section 7 Preventative Maintenance Mains voltages and d. The instrument must be disconnected from the mains supply before removing the covers.
If the mains lead requires attention, have it repaired by a qualified electrician. Very little maintenance is required other than standard laboratory good housekeeping procedures. The maintenance required is given in Table Operator selection of the following self tests; keyboard, display, metering pump, control valve.
Display of current software version and ability to reset default settings. Run 5 counts. Let sit for 5 min. Pergorm prime cycle with Isoton II. Very background count. System Flush-Perform a drain system function. Sit for 10 minutes. Drain system. Fill instrument with Isoton II. For the disinfection procedure refer to Appendix 7. In biological applications such as tissue culture, blood, etc. This procedure.
When no further instructions occur, the keyboard functionality has been satisfactorily checked. When the display returns to screen T1 the display has been.
During each test, one of the following status messages displays: Valve to fill position Wait for away sensor Resetting Valve Wait for home sensor. Note: Do not reset default settings unless advised by qualified Beckman Coulter personnel as all user settings will be lost.
Flushing an Aperture To flush particles or air bubbles away from the rear of the aperture proceed as follows: 1 Press. User Testing T1. User Testing list of tests. These messages are divided into the following categories: Status Messages.
These are informative messages that tell the operator what the instrument is doing and are part of normal operation. They appear at the bottom of the display, in inverse video. These warn the operator of any abnormal conditions requiring intervention.
These appear at the bottom of the display, in inverse video. The error message is continuously displayed, and the sounder is activated. The operator is allowed to exit the menu screen. Table 8. Number of particles too low for accurate results. Message only occurs during calibration. Can not occur during sample analysis. Informs operator flow rate is being measured. Informs operator aperture resistance is being measured. Aperture impedance and preamplifier impedance not matched.
Occurs when conductivity of electrolyte solution significantly changed without going back through Setup Menus. Is within noise level of 5 and 2. The rms root mean square value of electronic noise is measured independently of count. This is a warning. Sample analysis not inhibited as the resultant increase in count. Still makes count useful in some cases for comparison data. Alternately change to an Aperture Tube with smaller aperture.
So much noise being counted that results are unreliable. Consider using an Aperture Tube with smaller aperture and raising conductivity of electrolyte solution. Section 8 Status and Error Messages. INDEX error messages definition.
The information contained in this appendix is designed to help operators who are unfamiliar with these techniques. The relevance of each of the following procedures is dependent on the type of analysis to be performed. If a significant amount of sample lies outside the size range of one aperture diameter, more than one size of Aperture Tube may be required for a complete analysis.
In such a case, the determination of the largest and smallest particles present in the sample for analysis can be found by one of the following methods: a Microscopic inspection. If however, two or more apertures are suitable for an application, the larger size will complete the measurements more rapidly. A smaller aperture is more suitable for high concentration suspensions. Standard Aperture Tubes are defined as Aperture Tubes which have a 0.
Before mixing electrolyte solutions consider any possible risk. Some electrolytes are mutually incompatible and may generate gas or cause precipitation, if mixed. Always drain the Hydraulics System of old electrolyte solution before using a new or different one. It is essential to use an electrolyte solution compatible with the characteristics of the material being analyzed. An electrolyte solution must not change the size or dispersed state of the sample but must have sufficient conductivity to make the operation of the Z Series possible.
It is essential that the blank count of a diluent is statistically insignificant relative to the sample count. Also refer to Para A1. ISOTON II diluent is a filtered, phosphate-buffered saline solution compatible with human blood cells, and may be used for the suspension of most biological cells and many industrial samples. The composition of a diluent can be modified to take into account particular sample characteristics e. For biological samples a main consideration is the avoidance of the lytic properties in the diluent.
To help select a suitable electrolyte solution, Appendix 2 lists aqueous electrolytes, and Appendix 3 lists many of the more common particulate substances, together with most suitable electrolyte solution s. Filtration of Electrolyte Solutions It is essential that the background count from the electrolyte solution be as low as practicable refer to Table A This is achieved by careful filtration.
The following procedures are satisfactory for most laboratory-made diluents: Note:. If this material is presented in an agglomerated or flocculated form then an untrue count results. In some instances however, especially with airborne dusts, it is important to count the particles in an agglomerated state and dispersion to the ultimate particle size is undesirable.
In almost all cases, dispersion will be aided by the addition of a few drops of non-ionic dispersant such as Beckman Coulter Dispersant, obtainable from Beckman Coulter Particle Characterization, or its distributors. There are four usual methods of dispersion, which are as follows.
Gentle mixing of the powder with a drop or two of glycerol, alcohol, or suitable dispersant e. This is done on a watch glass, using a spatula.
Ultrasonic probes can cause fracture of some types of particles. This is rarely the case with baths for recommended models contact Beckman Coulter Particle Characterization or their authorized Distributors.
After gentle mixing as described above, a sub-sample of the material is placed in a clean beaker and diluted by stirring with a few drops of electrolyte solution. More electrolyte solution is added and the beaker placed in a small ultrasonic bath for 15 to 60 seconds, or until dispersion is complete. Simultaneous stirring with a clean glass rod reduces the agitation time required.
This method is the one generally found most satisfactory, but should not be used with fragile particles or biological cells.
When the particle surface is hard e. Allow air bubbles to disperse before starting the count. High-speed mixing can cause unwanted fracture of some particulate materials.
A wide range of electrolyte solutions may be required, dependent on the nature of the material and its solubility in water. Beckman Coulter Particle Characterization and its distributors maintain a detailed list of electrolyte solutions found suitable for several hundred materials and will be pleased to recommend electrolyte solutions for any given material, or assist in developing a suitable technique for new materials. A partial list is given in Appendix 3. Before using electrolyte solutions, check the stability of dispersion of the material.
After dispersion and mixing for one minute with the stirrer on the beaker platform, take several counts at the coarse end d max and the fine end d min of the size range. After the completion of the full analysis, or after l5 minutes further stirring, recount at the same levels. Table A shows what a comparison of the counts will indicate. Table A Stability of Dispersion d max.
Try different electrolyte solutions e. Presaturate electrolyte solution with sample before filtration. Use common-ion effect to depress solubility e. Deflocculation: Wait until stable. Use different electrolyte solution, dispersant, pH.
Use ultrasonics low power bath. A Table A Stability of Dispersion. Other specific notes are included where necessary. An electrolyte solution list cannot guarantee a perfect result; this also depends on sample preparation, dispersion, and even particle size - the finer the material the more reactive soluble, undispersible it will usually be.
This list, however, provides a starting point. There are very few materials which cannot be analyzed using the Beckman Coulter technique. Occasionally some organic compounds pesticides, drugs are too soluble in any electrolyte solution, and particles dispersed in mineral oils may require prior separation see list under Membrane Filters before size analysis.
Contact Beckman Coulter Inc. Always observe the usual chemical precautions; for example, do not mix strong oxidizing agents such as chromium trioxide, nitric acid, chlorates, perchlorates, or permanganates with glycerol.
Always check however, that this does not cause flocculation or poor dispersion. Usually, non-ionic dispersants are best for use with electrolyte solutions, e.
The optimum choice is largely a matter of trial and error. The amount of surfactant used should be small e. A dispersant can dissolve some organic materials e. Glycerol reacts slowly with phosphate electrolyte solutions. This may not be significant if only large particle sizes, e. Dispersion is best aided by low power ultrasonics baths - not probes, which can fracture some particles , and in some cases e.
The following list of electrolyte solutions has been provided by Beckman Coulter Particle Characterization laboratories from their more comprehensive records.
Beckman Coulter Particle Characterization provides an advisory service for customers with problems regarding materials not listed here.
CH2 OH. Often added to help suspend large particles, and to increase the viscosity of the electrolyte solution when using large apertures. Often added to help suspend large particles. De-gas first; ultrasonic agitation 2 minutes very suitable.
Sulframin has been used as dispersant; electrolyte solution has been presaturated with Boric Acid. Sodium chloride reacts. Difficult to wet mechanically, but ultrasonics with Coulter Dispersant very successful. In flake form it gives the characteristic streaming patterns in the stirred suspension, looking unhomogeneous, but this does not affect results. Teepol XL successful dispersant. Activated carbon will release adsorbed gases on passage through aperture giving spurious results.
Disperse powder in a little warm glycerol with spatula, add a little electrolyte solution and boil for a few minutes. Cool and add remainder of electrolyte solution; count. See CLAY. Add 0. Weight integration i. Correlation of results with other techniques e. Disperse mechanically, e. To measure particles in electrolyte solutions, e. Usually coarse and in very narrow size range, e. Solids or oil in water can be measured. Two-stage dilution of e. Stability of emulsion over analytical period must be checked and maintained.
Heat treated ferrites are magnetic and cannot be dispersed stably to unit crystals; the agglomerate size will be fairly stable and measurable. Preferably measure before heat-treatment. Fibers cut to constant known length can be measured for diameter the denier distribution.
Those of constant known diameter can be measured for length distribution, if they do not exceed 2 x diameter of aperture used. Coulter Dispersant has been used as dispersant. Saturate sample in electrolyte solution high concentration before. Reacts fairly quickly. This may be an effect of using too much aperture current. For most latices. Slow flocculation according to Smoluchowsky equation.
Useful for flocculation studies. Lead oxide is soluble in alkaline solutions. Disperse by spatulation e. Keep aperture voltage below some 6V to 12V.
Conductivity effects may be minimized by coating the surface of the particles with a quaternary ammonium compound, e.
High particle concentration; weight integration analysis method may be required. See under chemical name. See also under chemical name. Spatulate in glycerol or dispersant before dilution with electrolyte solution. Can cold-sinter within hours of milling to cause permanent agglomeration. Comments on, or additions to this list will always be welcomed by the Customer Support Laboratory,Beckman. A A3. Quick, accurate and satisfactory calibrations may be made with essentially monosized particles available from various sources.
The following materials Beckman Coulter calibration materials can be obtained from Beckman Coulter Particle Characterization Group for calibration by using the modal sizes. These particles are suspended at a concentration in distilled water containing surfactant and preservative such that one to five drops are required in approx.
Recommended concentrations are given with each vial, as are the various assayed sizes under different measurement conditions. Polystyrene Divinyl Benzene latexes are particularly recommended as they will not readily change size upon immersion in alcohols, ketones or any aqueous electrolyte solution. Redispersion of Calibration Particles Beckman Coulter calibration suspensions contain both a preservative and a surface active agent especially chosen to reduce any tendency of the particles to agglomerate or cake upon storage.
Before use, shake the vial vigorously for several seconds, and ensure that no sediment is left in the vial. Add one or more drops to the electrolyte solution as required. Further dispersion by ultrasonic agitation should not be necessary. Each pack of Beckman Coulter Particle Characterization Calibration Standards carries an assay sheet expressing median and modal diameters etc. The basic procedures used by Beckman Coulter Particle Characterization to assay these sizes are detailed by Harfield and Wood[ 1 ].
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