NucleoCounter ® NC-200 ™ Instrument User's Guide P/N 991-0203 Revision 1.
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NucleoCounter NC-200™ Flexible Cell Counter P/N 991-0203 (English) Revision 1.1 October, 2011 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: Sales@chemometec.com 991-0203 Rev. 1.1 Sep.
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Caution! This equipment must be operated as described in this User’s Guide and documents referred to herein. Please read the entire guide and referred documents before attempting to use this unit. Please pay attention to that gloves or protective clothing are not worn on the illustrations/pictures shown in this User’s Guide. However, ChemoMetec A/S does recommend that the user wear suitable protective clothing et c.
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Declaration of Conformity Declaration of Conformity Name of product: NucleoCounter NC-200 Type: Flexible Cell Counter Other identifying data: Part no. 900-0201 We declare under our sole responsibility that the products, to which this declaration relates, are in conformity with the Council Directives on the approximation of the laws of the EEC Member States relating to the following: Directives Electromagnetic Compatibility (2004/108/EC).
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WEEE directive information – Europe only WEEE directive information – Europe only Correct Disposal of This Product (Waste Electrical & Electronic Equipment) - Europe only This marking shown on the product or its literature, indicates that it should not be disposed together with other household wastes at the end of its working life.
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Introduction and intended use Introduction and intended use The NucleoCounter NC-200 is a compact cell-counter intended to be used for precise cell counting within the life-science and pharmaceutical markets. The NucleoCounter NC-200 system is intended for research use only, not for diagnostic use. The NucleoCounter NC200 uses disposable cassettes from ChemoMetec A/S.
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Warnings and precautions Warnings and precautions Whenever the symbol appears on the NucleoCounter NC-200 instrument, it indicates that the manual must be consulted for precautions and warnings. Power and cables Use the shielded USB cable supplied with the NucleoCounter NC-200 to ensure that appropriate EMI classification is maintained for the intended environment. The USB interface connector of the NucleoCounter NC-200 must only be connected to SELV circuits.
Warnings and precautions Electromagnetic interference NOTE: This equipment has been tested and found to comply with the limits for a Class B digital device, pursuant to Part 15 of the FCC Rules. These limits are designed to provide reasonable 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 instructions, may cause harmful interference to radio communications.
Table of contents Table of contents DECLARATION OF CONFORMITY ...................................................................................... VII WEEE DIRECTIVE INFORMATION – EUROPE ONLY .............................................................. IX INTRODUCTION AND INTENDED USE ................................................................................ XI WARNINGS AND PRECAUTIONS ......................................................................................
Table of contents 8.3 9 P OWER - ON FAILURE ............................................................................................... 19 TECHNICAL SPECIFICATIONS ..................................................................................... 21 9.1 T HE N UCLEO C OUNTER NC-200 ................................................................................ 21 9.2 S AMPLE MEDIAS .................................................................................................... 21 9.
1 Inspection, Unpacking and Installation of Equipment 1 Inspection, Unpacking and Installation of Equipment Upon receiving the order from ChemoMetec A/S, the box or boxes should be carefully inspected for any damage that may have occurred during shipping. Any damage must be reported to the carrier and to ChemoMetec A/S immediately. Unpack the order, saving the packing materials for possible later use.
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2 Cell counting with the NucleoCounter NC-200 2 Cell counting with the NucleoCounter NC-200 The NucleoCounter NC-200 is a bi-color fluorescent cell-counting instrument. The NucleoCounter has the advantage that only short training time is required before the user can run highly advanced cell counting protocols. The instrument itself is calibration free. The individual calibration parameter for a cassette will be read and used by the instrument during analysis.
2 Cell counting with the NucleoCounter NC-200 2.1.1 Fluorescence microscope The NucleoCounter NC-200 is a fluorescent microscope with a magnification of approximately x1.3. It is equipped with 3 light sources (2 for fluorescent use and one for dark-field analysis). It has a dual-band emission filter. At present only Via1-Cassette is used the NucleoCounter NC-200. A number of different protocols can be run on the NucleoCounter NC-200.
2 Cell counting with the NucleoCounter NC-200 2.2 The Cassette Figure 2 The Via1-Cassette. Here fluorescent dyes AO (acridine orange) and DAPI (4',6 diamidino-2-phenylindole) are immobilized in the first part of the flow system. 2.2.1 Loading the Cassette The cassette is loaded by gently pressing the white piston down to the top of the cassette, which creates a partial vacuum in the flow system.
2 Cell counting with the NucleoCounter NC-200 The cassettes to be used with NucleoCounter NC-200 are highly specialized devices, which hold the key to several of the unique features of the NucleoCounter NC-200 concept. For instance in the Via1-Cassette the fluorescent dyes AO (acridine orange) and DAPI (4',6diamidino-2-phenylindole) are immobilized inside the flow channels of the disposable cassette.
3 Key, LED indicator and PC control 3 Key, LED indicator and PC control 3.1 Interactive Controls Figure 4 The front of the NucleoCounter NC-200 The figure above shows the front of the NucleoCounter NC-200. Two operational areas are present: Lid with cassette fixture behind Run button and a Tri color status LED indicator The lid is located in the first area where the sample, contained in a cassette is placed. When the lid is opened, a cassette can be placed in the fixture.
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4 Start-up 4 Start-up 4.1 Power on, power off On the rear of the instrument, two plugs and one fuse holder are located. When seen from the rear the USB 2.0 port is to the left and the fuse holder and DC Pow er Plug is at the right. The NucleoCounter NC-200 is powered by a 24VDC external power supply. To connect the power supply to the NucleoCounter, connect it to the DC Power Plug (see Figure 5), and plug the mains power cord of the power supply into a wall outlet. Figure 5.
4 Start-up 4.1.1 Starting up The NucleoCounter NC-200 is connected to a PC by inserting the respective plugs of a USB cable into the instrument and the PC. Once the NucleoView NC-200 program is launched the stepping motor for the piston actuator performs a homing procedure in order to assure correct positioning of the actuator. After this initialization, the status indicator on the front will turn green and the NucleoCounter NC-200 is ready for operation. 991-0203 Rev. 1.1 Sep.
5 Operation of the NucleoCounter 5 Operation of the NucleoCounter 5.1 Ready mode Once the NucleoCounter NC-200 and the NucleoView NC-200 software are ready, analysis can begin. In the NucleoView NC-200 software, the browse menu can be used to select the protocol to be performed. Once selected, the run button on the front of the instrument (see Figure 4) or the run button in the software can be used to execute the analysis. Browse for protocol Figure 8 NucleoView NC-200 5.
5 Operation of the NucleoCounter Figure 9. Illustration of how to hold and place the cassette in the fixture 5.2.1 Actuator movements When a cassette is analyzed after Run is pressed, the piston actuator will move the piston rod down through the cylinder of the cassette. Hereby, the sample material is transported to the cassette measurement chamber. The form and dimensions of the flow channels facilitate effective mixing of the sample. Sensors monitor sample flow.
5 Operation of the NucleoCounter 6 NucleoView NC-200 Figure 10 NucleoView NC-200 with a typical result after analysis The NucleoView NC-200 software controls the NucleoCounter NC-200 instrument and image analysis of the recorded images. The program is installed on a Windows PC (Windows XP SP3 or Windows 7). Several options for data presentation are available in the NucleoView NC-200 software. The main results of the analysis are presented directly on the user interface of the software.
5 Operation of the NucleoCounter The events registered as cells can (when enabled in the protocol) be inspected by launching the Protocol Adaption Editor in the Tools menu and selecting to show events included in the count prior to running an analysis. This will launch a plot manager that shows a scatter plot with a gate that encloses the events recorded as cells. Furthermore, two histograms will be displayed showing the intensity and area of the events in the picture.
7 Maintenance of NucleoCounter NC-200 7 Maintenance of NucleoCounter NC-200 7.1 Cleaning 7.1.1 Instrument enclosure. Depending on the environment in which the NucleoCounter NC-200 is operated, it is suggested that regular cleaning of the enclosure is carried out. When cleaning the enclosure it is recommended to use a soft moist cloth and gently wipe the surface. Any contamination, which does not come off immediately, should be rubbed gently wit h a cloth wetted with mild detergent.
7 Maintenance of NucleoCounter NC-200 7.1.3 Spill of liquid If liquid has been spilled on the instrument it can contaminate elements of the optical system. This contamination can show up in the image in several 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 of the NucleoCounter NC-200.
8 PC- maintenance, Error messages and Power-on failure 8 PC- maintenance, Error messages and Power-on failure 8.1 Required hard disk space for data storage. The CM files stored on the hard disk can vary from 6MB to 30MB. Therefore it is important from time to time to check whether enough free space is present on the hard disk. ChemoMetec recommends a minimum of 10G Bs free space on the hard disk. It is recommended to make a periodic archiving of CM files to external storage.
8 PC- maintenance, Error messages and Power-on failure Figure 14. Location and removal of the fuse Disconnect the power supply from the NucleoCounter. Then remove the fuse holder using a screwdriver as indicated in Figure 14 (turn counter clock wise). Inspect the fuse. If it needs replacement use one as described by ChemoMetec A/S.
9 Technical specifications 9 Technical specifications 9.1 The NucleoCounter NC-200 Optics Lens with x1.3 magnification, 1/2.” CMOS with 2560 x 1920 pixels Sample device NC-200 Cassettes, Excitation (nm) 2 LED light sources with peaks at 365 nm, 505 nm and dark field light Emission (nm) A single dual-band emission filter: 410-460 nm and 540-650 nm Sample consumption 60 μl Analyzed volume 1.4 μl sample Optimal Range 5 x 10 - 1 x 10 cells/ml Cell types Mammalian cells.
9 Technical specifications Directives Electromagnetic Compatibility (2004/108/EC).
10 Equipment and Accessories 10 Equipment and Accessories On www.chemometec.com an updated list of Equipment and Accessories for the NucleoCounter NC-200 are available. 991-0203 Rev. 1.1 Sep.
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10 Equipment and Accessories Appendix A: Description of changes from latest revision This chapter describes changes from latest revision of this User’s guide. 991-0203 Rev. 1.1 Sep.
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Appendix B: WEEE directive information in more EU languages Appendix B: WEEE directive information in more EU languages United Kingdom Correc t Disposa l of This Product (Waste Electr ical & Elec tronic Equ ipment) - Europe only This marking show n on the product or its lite rature , indicates that it should not be dispo sed t ogethe r w ith othe r household wastes at the e nd of its work ing life.
Appendix B: WEEE directive information in more EU languages Norway Korrekt avhending av dette produkt (Avfall elektrisk og elektron isk utstyr) - Kun Europa Denne merk ingen som vises på produktet elle r dens dokument asjon, indikere r at de n ikke skal kastes sammen med annet hus holdni ngsavfall ved slutten av sin levet id.
Appendix B: WEEE directive information in more EU languages Finland Tämän tuotteen turvallinen hävittämin en (elektroniikka ja sähkölaitteet) - Vain Eu rooppa Oheinen me rkintä tuotteessa tai tuotteen ohe ismate ria alissa me rkitsee , että t ätä t uotetta e i tule hävittää kot italousjättee n mukana sen e linkaa ren päätyttyä. Hallitsemattomasta jätteenkäs ittelystä ympä ristölle ja kanssa ihm isten te rveydelle aiheut uvien vahink ojen välttäm iseksi tuote tulee kä s itellä muista jätteistä e rillään.
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Appendix B: WEEE directive information in more EU languages The information contained herein is to the best of our knowledge accurate and complete. However cell species and cell environments may vary in property. Therefore systematic and/or random deviation between estimates obtained by the NucleoCounte r and other cell counting methods may occur.
