ChemoMetec NucleoCounter Page 3 User Manual Download

Page: 3 / 76

Nu cl e o Cou n t er



Mammalian Cell Counter

P / N 9 9 1 - 0 0 0 9 ( E n g l i s h )

R e v i s i o n 1 . 3

J u n e , 2 0 1 5

ChemoMetec A/S

Gydevang 43, DK-3450 Allerød, Denmark

Telephone: (+45) 48 13 10 20, Fax: (+45) 48 13 10
21

Internet: www.chemometec.com,

E-mail:[email protected]

Summary of Contents for NucleoCounter

Page 1: ...NucleoCounter User s Guide P N 991 0009 Revision 1 4 Technology that counts...

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Page 3: ...ll Counter P N 9 9 1 0 0 0 9 E n g l i s h R e v i s i o n 1 3 J u n e 2 0 1 5 ChemoMetec A S Gydevang 43 DK 3450 Aller d Denmark Telephone 45 48 13 10 20 Fax 45 48 13 10 21 Internet www chemometec co...

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Page 5: ...ssistance with NucleoCounter or the NucleoView software to place an order for a NucleoCounter or consumables call 45 48 13 10 20 fax 45 48 13 10 21 or send e mail to sales chemometec com Disclaimer No...

Page 6: ...v This page was intentionally left blank...

Page 7: ...al equipment for measurement control and laboratory use EMC requirements emission and immunity Annex B and C from A1 1998 is used This device complies with Part 15 of the FCC Rules Operation is subjec...

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Page 9: ...prevent possible harm to the environment or human health from uncontrolled waste disposal please separate this from other types of wastes and recycle it responsibly to promote the sustainable reuse o...

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Page 11: ...rs The NucleoCounter is developed as a stand alone instrument but optionally it can be connected to a computer via an USB interface When connected to a computer the NucleoViewTM software offers variou...

Page 12: ...fficient cell counting and viability determinations of samples containing mammalian cells The NucleoCounter NucleoCassette the Lysis buffer and the Stabilizing buffer are not intended for human or vet...

Page 13: ...Introduction and intended use xii This page was intentionally left blank...

Page 14: ...ounter has to comply with the standards UL 1950 and IEC EN 60950 Table 1 Description of the NucleoCounter USB interface connector Use the shielded printer cable supplied with the NucleoCounter to ensu...

Page 15: ...please follow the instructions for connecting the power supply The NucleoCounter shall only be used with one of the following external power supplies Power supply Class I Proton Electronic Industrial...

Page 16: ...able protection against harmful interference in a residential installation This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the ins...

Page 17: ...ective clothing eye protection and gloves must be worn when using potentially hazardous liquids General Any biological specimen should be handled as if it is capable of transmitting infectious disease...

Page 18: ...GENT A100 AND STABILIZING BUFFER REAGENT B 9 3 CONTROL BUTTONS KEYPAD AND DISPLAY 12 3 1 INTERACTIVE CONTROLS 12 3 1 1 Control Buttons 12 3 1 2 Keypad 13 3 2 INTERFACE DISPLAY 13 3 2 1 Status indicato...

Page 19: ...liquid 35 8 TROUBLESHOOTING ERROR MESSAGES 37 8 1 NO VALID CASSETTE 37 8 2 ANALYSIS ABORTED 37 8 3 ACTUATOR ERROR MESSAGES 38 8 4 SAMPLE COULD NOT BE ANALYZED 38 8 5 SENSOR ERROR 39 8 6 POWER ON FAILU...

Page 20: ...e must be reported to the carrier and to ChemoMetec A S immediately Unpack the order saving the packing materials for possible later use Also be sure to save the User s Guide for instruction and refer...

Page 21: ...1 Inspection and Unpacking of Equipment 4 This page was intentionally left blank...

Page 22: ...cell suspension please refer to appropriate Application Notes provided by ChemoMetec concerning stabilizing of the lysed cell suspension Basics of the NucleoCounter system is explained on the next fe...

Page 23: ...roscope Before the NucleoCounter fluorescence microscope is activated a NucleoCassette is loaded with sample and inserted into the insertion slit of the instrument The lid is closed in order to exclud...

Page 24: ...the CCD chip of the camera A fluorescent image of the content of the measurement chamber is thus achieved Example of such an image is shown in Figure 4 Each spot in the image represents a cell contain...

Page 25: ...the piston the flow system containing the fluorescent dye propidium iodide and the measurement chamber Figure 5 The NucleoCassette The fluorescent dye propidium iodide is immobilized in the first part...

Page 26: ...of the measurement chamber of the cassette The dot code is read and decoded in the instrument during analysis and the volume analyzed is determined by multiplying the imaged area with the depth of the...

Page 27: ...the Stabilizing buffer is to raise pH of the sample mixture thereby optimizing the fluorescence of propidium iodide and to stabilize the cell nuclei The combination of treating the cell sample first...

Page 28: ...Fejl Henvisningskilde ikke fundet 11 This page was intentionally left blank...

Page 29: ...ent see Figure 6 Table 3 Control Buttons on NucleoCounter On Off Use this button to turn the instrument on and off Run Use this button to start measuring Esc Use this button only as instructed in this...

Page 30: ...On Off button see Figure 6 illuminates when the unit is turned on It is green when the instrument is ready to measure and it turns red while the instrument is measuring or displaying an error message...

Page 31: ...ounter The NucleoCounter can now be turned on and off by pressing the On Off button on the front panel 4 1 1 Starting up Caution During start up the NucleoCounter performs a self check Never insert or...

Page 32: ...green NucleoCounter Ready Figure 9 The NucleoCounter is ready for cell counting If the instrument has performed as described above it is ready for use If the instrument displays an error message at s...

Page 33: ...covers the cassette insertion area This lid is designed to protect the area from dust and other potential contaminants and to keep external light from interfering with the fluorescent image recording...

Page 34: ...ring The Run button When a cassette has been loaded and inserted into the instrument the measurement is activated by pressing the Run button 5 3 1 Actuator movements When Run is pressed the actuator m...

Page 35: ...e complete process described above may be interrupted Please familiarize yourself with possible error handling of the NucleoCounter presented in chapter 8 5 4 Result display Following a successful ana...

Page 36: ...ults follow the directions below a letter or number enclosed with and corresponds to a button on the keyboard Press F 2 1 on the keyboard of the NucleoCounter Press enter Press the mode number of the...

Page 37: ...printer port Please refer to Technical Note 001 Part No 994 0013 with respect to setting and using the Time and Date features 5 4 4 Reset Counter F30 The function F30 F 3 0 is used to reset the Count...

Page 38: ...w LCD contrast value can now be chosen Contrast 6 1 Up 2 Down store Figure 16 LCD contrast value 6 is going to be chosen Press to store the chosen value or Esc to return to the previous value If the v...

Page 39: ...s the number of CCD pixels that show elevated signal levels If the number of pixels is above a predefined limit then the cassette insertion slit can be contaminated with an interfering particle like a...

Page 40: ...e Zero Count Check If no contamination of the insertion slit can be detected an OK message appears in the display see the figure below Press any key to return to Ready mode 0 count OK Press any key Fi...

Page 41: ...No 004 How to clean the NucleoCounter Perform the Zero Count Check again If the error appears even after several attempts to clean the insertion slit with compressed air and if necessary the NucleoCo...

Page 42: ...5 Operation of the NucleoCounter 25 This page was intentionally left blank...

Page 43: ...e plasma membranes of all the cells must be disrupted to render all the nuclei susceptible to staining with propidium iodide The disruption is achieved by treatment with a Lysis buffer4 Reagent A100 f...

Page 44: ...e vial upside down 5 10 times Alternatively you can use the pipette Add 200 l of the Stabilizing buffer and mix thoroughly This mixture is called the Stabilized lyzate Load the NucleoCassette with the...

Page 45: ...puter if one is connected Examine the result of the analysis The measurement range of the NucleoCounter is 5 103 2 106 cells ml If the cell concentration of the stabilized lyzate is outside the measur...

Page 46: ...al cell concentration The result of the NucleoCounter should be regarded as reliable in these cases but the operator must be aware that a high relative error is associated with these results due to th...

Page 47: ...Mnv is 1 The result of the count of non viable cells Cnv is 3 0 104 cells ml The calculation of viability is shown below 98 5 100 3 10 6 7 1 10 3 0 3 10 6 7 100 M C M C M C viability 5 4 5 t t nv nv...

Page 48: ...this method to dilute a cell suspension take out a sample of the diluted cell suspension and add Lysis buffer and Stabilizing buffer in the normal way Finally calculate the multiplication factor Examp...

Page 49: ...6 Total and viability count 32 This page was intentionally left blank...

Page 50: ...nst dust and other contaminants Therefore great care must be taken to ensure that the lid covering the cassette insertion area is closed when cassettes are not being loaded into or removed from the Nu...

Page 51: ...g dust particles will normally not influence the counting of cells The NucleoCounter will distinguish between cells and contaminants since cell is generally significantly smaller than contaminants Dus...

Page 52: ...ways and have many causes but a common feature is that it cannot be removed by the use of compressed air Even though such phenomena are clearly visible on the image it only rarely affects the results...

Page 53: ...7 Maintenance of NucleoCounter 36 This page was intentionally left blank...

Page 54: ...r message displayed when the NucleoCounter does not recognize the cassette as valid This error message is displayed in the following situations The cassette has been analyzed previously The cassette i...

Page 55: ...r of the error messages appears Actuator error Refer to manual Press any key Figure 29 The NucleoCounter can display this message regarding the actuator First examine if the cassette has been inserted...

Page 56: ...sors has a faulty operation an error message will be displayed see Figure 31 Sensor error Refer to manual Press any key Figure 31 This error message is displayed if one of the sensors which read the b...

Page 57: ...Then remove the fuse holder by use of a screwdriver turn CCW as indicated in Figure 32 Inspect the fuse If it needs replacement use a fuse 1 25A 250Vac UL 248 If the NucleoCounter does not turn on aft...

Page 58: ...8 Troubleshooting Error messages 41 This page was intentionally left blank...

Page 59: ...pressing Run on the NucleoCounter the result will be displayed within 30 seconds Operation Menu controlled by means of keyboard and LCD display Physical data Weight Height Width Depth 3 kg 26 cm 38 cm...

Page 60: ...ealed foil bag at max 30 C Stability See documentation delivered with the NucleoCassettes 9 3 EMC EMI standards The NucleoCounter complies with EMC EMI standards as follows EN61326 1997 A1 1998 Class...

Page 61: ...A 500 ml 910 0001 A reagent for lysis and disagregation of cells Reagent A100 500 ml 910 0003 A reagent for lysis and disagregation of cells Reagent B 500 ml 910 0002 A stabilizing buffer Reagent C 5...

Page 62: ...r Reagent A100 or Reagent B Bottle Stand 929 0001 Holder for Reagent A100 and Reagent B bottles Thermo Printer PCNEOS S2BN 939 0006 Optional thermo printer using thermo paper for the NucleoCounter LTS...

Page 63: ...VT rated min 60C 18 AWG 3 conductors Provided with molded on grounding type NEMA 5 15P attachment Plug rated min 125 Vac min 2 5A Opposite end terminates in molded on IEC320 style connector rated min...

Page 64: ...ied by the manufacturer the protection provided by the NucleoCounter and the external power supply may be impaired Contact ChemoMetec A S for information on order of a specific Power supply cord liste...

Page 65: ...is located just below the USB connector on the rear side as apparent from Figure 34 and Figure 35 Figure 34 Location of the Printer output on the NucleoCounter Figure 35 Connecting the Printer cable...

Page 66: ...6 NucleoCounter Test Kit Figure 37 The NucleoCounter test kit The NucleoCounter test kit consists of 3 levels of bead concentration and a blank The test kit can bu used to test the performance of a Nu...

Page 67: ...Figure 38 NucleoView The NucleoView PC software is used for handling of the obtained cell counts The program is installed on a Windows PC Win2000 Win98 2 edt WinME WinXP A USB driver and the program...

Page 68: ...5 1 35 x 10E5 1 3 x 10E5 9 13 x 10E5 9 37 x 10E5 9 25 x 10E5 1 36 x 10E5 1 45 x 10E5 1 4 x 10E5 9 38 x 10E5 9 62 x 10E5 9 50 x 10E5 1 46 x 10E5 1 55 x 10E5 1 5 x 10E5 9 63 x 10E5 9 87 x 10E5 9 75 x 10...

Page 69: ...10E5 1 876 x 10E6 1 925 x 10E6 1 90 x 10E6 4 91 x 10E5 5 12 x 10E5 5 00 x 10E5 1 926 x 10E6 1 975 x 10E6 1 95 x 10E6 5 13 x 10E5 5 37 x 10E5 5 25 x 10E5 1 976 x 10E6 N A 2 00 x 10E6 5 38 x 10E5 5 62...

Page 70: ...F11 The appropriate setting is selected by the user function F11 F11 on the keypad Press 1 to toggle up and 2 to toggle down through the menu For firm ware 4 10 the options are Type 1 7 For firm ware...

Page 71: ...e 1 is the least sensitive default NC 100 and should be used for the counting of mammalian cells We recommend that NucleoView is used to select the best Type for your cells by ensuring that cell signa...

Page 72: ...12 Appendix 2 Extended NucleoCounter NC 100TM Settings 55 This page was intentionally left blank...

Page 73: ...drugimi gospodarskimi odpadki Slovakia Spr vna likvid cia tohoto v robku Elektrotechnick a elektronick odpad Plat len pre Eur pu Toto ozna enie na v robku alebo v sprievodnej bro re hovor e po skon en...

Page 74: ...tliekomis Kad b t i vengta galimos nekontroliuojamo atliek i metimo alos aplinkai arba moni sveikatai ir siekiant skatinti aplink tausojant antrini aliav panaudojim pa om atskirti j nuo kit r i atliek...

Page 75: ...o no debe eliminarse mezclado con otros residuos comerciales Greece Germany Korrekte Entsorgung dieses Produkts Elektrom ll Nur Europa Anzuwenden in den L ndern der Europ ischen Union und anderen euro...

Page 76: ...results obtained from use of the NucleoCounter or NucleoCassette shall be construed to imply any warranty or guarantee ChemoMetec A S and affiliated companies shall not be held liable for damages and...

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