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2 Introducing the NucleoCounter  

  12 

 

Figure 5

 The NucleoCassette. The fluorescent dye, propidium iodide, is immobilized 

in the first part of the flow system.   

 

2.2.1  Loading the cassette 

The NucleoCassette is loaded by gently pressing the white piston,  which creates a 

partial vacuum in the flow system. The tip of the NucleoCassette must be immersed 

into a lyzate mixture when pressing the piston, resulting in lyzate being loaded into 

the flow system. Approximately 50µl is loaded into the flow system.  

2.2.2  Fluorescent dye 

The fluorescent dye, propidium iodide, is immobilized in the first part of the flow 

system. As the stabilized lyzate is loaded into the NucleoCassette, the immobilized 

propidium iodide is dissolved immediately and mixed with the lyzate. Propidi um 

iodide intercalates with DNA and forms a fluorescent stain, absorbing green light 

and emitting red light, which is utilized for detection of stained cells. Furthermore 

PI's ability to fluoresce is enhanced significantly upon binding to DNA thereby 

substantially enhancing selectivity.   

During the analysis the stained mixture is transported through the flow system, 

where it is mixed with the propidium iodide, towards the chamber where the actual 

cell counting is performed. The precise volume analyzed, is d etermined individually 

for each cassette during analysis. This volume is approximately 1 µl, and thus the 

volume analyzed is 10-20 times that of a conventional hemacytometer. After 

analysis the NucleoCassette is disposed of as biological waste.  

2.2.3  Sample volume 

During production each cassette is marked with a dot code, which specify the 

precise depth or thickness of the measurement chamber of the cassette. The dot 

code is read and decoded in the instrument during analysis and the volume 

Summary of Contents for 900-0300

Page 1: ...NucleoCounter YC 100 User s Guide Revision 1 3 Technology that counts...

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Page 3: ...Counter P N 9 9 1 0 3 0 0 E n g l i s h R e v i s i o n 1 2 D e c e m b e r 2 0 0 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 chemome...

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Page 5: ...For sales assistance with NucleoCounter YC 100 or the NucleoView software to place an order for a NucleoCounter YC 100 or consumables call 45 48 13 10 20 fax 45 48 13 10 21 or send e mail to sales ch...

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

Page 7: ...ipment 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 subject to t...

Page 8: ...vii This page was intentionally left blank...

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: ...d of fluorescence microscopy The system enables the user to stain and count cells with good protection against being exposed to potentially hazardous DNA staining dyes The NucleoCounter YC 100 is deve...

Page 12: ...Introduction and intended use xi This page was intentionally left blank...

Page 13: ...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 ensure that you ma...

Page 14: ...Industrial Co Ltd model SPN 260 12 input rated 100 240 Vac 50 60Hz 1 6 A Output rated 12 Vdc 4 3 A Power Supply Class I Powerbox Europe AB model EBH 03 131 input rated 100 240 Vac 47 63 Hz 0 8A Output...

Page 15: ...off and on the user is encouraged to try to correct the interference by one or more of the following measures Reorient or relocate the receiving antenna Increase the separation between the equipment a...

Page 16: ...4 Never pipette by mouth Avoid cross contamination of the samples when preparing of the samples T his can compromise the quality of the results...

Page 17: ...handling 13 2 3 LYSIS BUFFER 13 3 CONTROL BUTTONS KEYPAD AND DISPLAY 15 3 1 INTERACTIVE CONTROLS 15 3 1 1 Control Buttons 15 3 1 2 Keypad 15 3 2 INTERFACE DISPLAY 16 3 2 1 Status indicator 16 3 2 2 Di...

Page 18: ...1 2 Optical elements 29 7 1 3 Removal of dust particles 30 7 1 4 Spill of liquid 31 8 TROUBLESHOOTING ERROR MESSAGES 33 8 1 NO VALID CASSETTE 33 8 2 ANALYSIS ABORTED 33 8 3 ACTUATOR ERROR MESSAGES 34...

Page 19: ...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 20: ...1 Inspection and Unpacking of Equipment 8 This page was intentionally left blank...

Page 21: ...cells Basics of the NucleoCounter system is explained on the next few pages The NucleoCounter system identifies and counts individual cell containing stained DNA The sample pre treatment is intended...

Page 22: ...gh the cylinder of the cassette Hereby the stained lyzate mixture is transported throug h the flow system of the cassette The form and dimensions of the flow channels facilitate effective mixing of ly...

Page 23: ...s known the cell density in the initial sample can be calculated The concentration of cells in the initial sample is presented in the NucleoCoun ter display If the optional software NucleoView is used...

Page 24: ...tes with DNA and forms a fluorescent stain absorbing green light and emitting red light which is utilized for detection of stained cells Furthermore PI s ability to fluoresce is enhanced significantly...

Page 25: ...windows of the cassette as clean as possible Caution In order to avoid contaminating the measurement window it is important not to touch the window when handling the cassettes Be careful when attempti...

Page 26: ...Fejl Henvisningskilde ikke fundet 14 This page was intentionally left blank...

Page 27: ...he instrument on and off Run Use this button to start measuring Esc Use this button only as instructed in this manual Del Use this button only as instructed in this manual Figure 6 Control Buttons Key...

Page 28: ...strument is ready to measure and it turns red while the instrument is measuring or displaying an error message The Status Indicator turns green after successful measurement when the results are displa...

Page 29: ...utton on the front panel 4 1 1 Starting up Caution During start up the NucleoCounter performs a self check Never insert or remove any cassette during the start up During start up a brief noise is hear...

Page 30: ...unting If the instrument has performed as described above it is ready for use If the instrument displays an error message at start up please refer to chapter 7 4 1 2 Shutting down Pressing the On Off...

Page 31: ...nt lid 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 rec...

Page 32: ...ents When Run is pressed the actuator moves the piston rod down through the cylinder of the cassette Hereby the sample dye mixture is transported to the cassette measurement chamber The form and dimen...

Page 33: ...sentations of the results In the default display mode mode 3 the result is displayed with number of digits corresponding to full representation of the number of cells per l with 2 3 or 4 digits An ove...

Page 34: ...ctively This feature is only used when a printer or a PC is connected to the printer port Please refer to Technical Note 001 Part No 994 0013 with respect to setting and using the Time and Date featur...

Page 35: ...trument in line 1 of the display and the instrument type in line 2 of the display The version number and date year month date of the integral software of the instrument is shown in line 3 The serial n...

Page 36: ...oCounter instrument using different sensitivity settings Figure 19 Object Intensity Histogram using different sensitivity settings The figure above illustrate firstly on top a sample measured with too...

Page 37: ...s usually observed as an object which is stationary in all images To initiate a Zero Count Check type in F50 Then the following text will appear in the display 0 count check Continue Esc Abort Figure...

Page 38: ...sertion slit is detected an error message is shown 0 count ERROR Refer to manual Press any key Figure 25 The Zero Count Check did not pass the test If the Zero Count Check returns an error it is recom...

Page 39: ...nded cells under conditions where viability of the cells is retained only cells directly stained with PI are assumed to be non viable Therefore when determining the concentration of non viable cells t...

Page 40: ...tion shown below 100 M C M C M C viability t t nv nv t t viability The percentage of viable cells in the original cell suspension Ct The total concentration of cells in the NucleoCassette the displaye...

Page 41: ...st 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 Nuc...

Page 42: ...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 43: ...ways and have many causes but a common feature is that it cannot be removed b y the use of compressed air Even though such phenomena are clearly visible on the image it only rarely affects the results...

Page 44: ...7 Maintenance of NucleoCounter 32 This page was intentionally left blank...

Page 45: ...cassette is not inserted properly into the NucleoCounter There is no cassette inserted into the NucleoCounter 8 2 Analysis aborted The NucleoCounter displays the error message shown in Figure 29 when...

Page 46: ...corrected 8 4 Sample could not be analyzed When the NucleoCounter cannot determine the concentration of cells during analysis the error message in Figure 31 is displayed Error Sample could not be ana...

Page 47: ...connected to the NucleoCounter and a working power plug If the NucleoCounter still does not turn on inspect and or replace the fuses as described below The fuse holder is located just above the Nucleo...

Page 48: ...8 Troubleshooting Error messages 36 This page was intentionally left blank...

Page 49: ...ion in the original cell suspension the result must be multiplied with the appropriate multiplication factor to compensate for any dilution Operation Menu controlled by means of keyboard and LCD displ...

Page 50: ...lows EN61326 1997 A1 1998 Class B A2 2000 Electrical equipment for measurement control and laboratory use EMC requirements emission and immunity Annex B and C from A1 1998 is used This device complies...

Page 51: ...e used for transmission of image data from the NucleoCounter to an external PC Power supply See section below A device for generating the DC voltage for the NucleoCounter from the mains supply Fuse 5x...

Page 52: ...ounter shall only be used with one of the following external power supplies Power supply Class I Proton Electronic Industrial Co Ltd model SPN 260 12 input rated 100 240 Vac 50 60Hz 1 6 A Output rated...

Page 53: ...50Vac Cord type min H05RR F or min H03VV F or min H03VVH2 F rated min 60C 2x0 75mm2 Provided with molded on un grounding type attachment Plug rated min 250 Vac min 2 5 A Opposite end terminates in mol...

Page 54: ...d with a printer output port It 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 3...

Page 55: ...43 2 1 3 Metal E nclosure Figure 36 Printer Output connector seen from the cable entry side...

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Page 57: ...u gimi 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 58: ...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 59: ...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 europ...

Page 60: ...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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