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Frequently asked questions |
FAQs—BandView®
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When I captured the Metaphase, some chromosomes were not captured. How can I add them?
Using ASI's Mergeview, all chromosomes not captured in the first acquisition can be added at a later stage. No need to recapture the metaphase again.
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Can I annotate my images?
Annotation of any image can be done in Metaphase or karyotype mode
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FAQs—Multi-Species
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Can I karotype new species with your system?
With the addition of the MultiSpecies module you can create forms or templates for Karyotyping any species.
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I work with chromosomes from rare species. Can I use BandView?
With the addition of the MultiSpecies module, all species can be karyotyped
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FAQs—FISHView
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What type of probe kits should I use for FISH?
You can use any commerical kit for FISH assays. Applied Spectral Imaging produces Whole Chromosome Painting kits.
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Which Fluorochrome filters do I need for my application?
The Fluorescence filters in the microscope are used to select the appropriate spectral signal emitted from your specimen. Before we are able to advise you on which filters to purchase, we need to know which application and probe kits you wish to use.
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FAQs—SkyPaint® Probes
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What is the content of vials no. 2–4?
Vial 2 contains a blocking solution to reduce the background, This step is optional and can be omitted from the process. Vials 3 and 4 contain antibodies conjugated to Cy5 and Cy5.5.
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What can be done to improve my results if the Cy dyes are weak?
You can repeat the detection step. Carefully remove the coverslip and wash the slides for 5 minutes in 4XSSC/).1%Tween 20. Continue with step 5 in the detection step.
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For how long can I keep the kit?
At the proper temperature of 4°C, the kit is stable for six months. (Vial no. 1 can be stored at –20°C). After this period, the antibodies in vials 2–4 should be replaced. The SkyPaint-DNA kit (not includingt vials 3 and 4) and the CAD (concentrated antibodies) kit is stable for nine months if stored at the proper temperature of –-20°C.
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Can I use other types of pretreatment?
Yes, Proteinaze K can be used. Brief incubation in Trypsin, at 1/5 –1/2 of the concentration that is used for G-banding, is also very useful for removing residual cytoplasm.
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Can I use a rapid wash without Formamide?
Yes, you can wash your SKY slides in 0.5XSSC for five minutes at 72°C and then for 2 min at 4XSSC/0.1% Tween20.
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Can I hybridize my slides for only 16–24 hours?
Yes, you can hybridize your SKY slides for only 16–24 hours and usually get very good results.
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Can I denature the slide and the probe together (codenaturation) ?
Yes, you can use codenaturation for the slide and the probe. In this case, there is no pretreatment.
Protocol for codenaturation:
1. Place10 ml from the probe on the slide, apply a glass cover and seal with rubber cement.
2. Place on a hotplate at 75°C for 1.5 minutes.
3. Transfer immediately to the incubator at 37°C for 16–24 hours.
Continue with the washing step.
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FAQs—Case Data Manager
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In our institute we would like to connect the ASI products to our patient database. Is this possible?
ASI already connects to several customer databases. However ,should this not be one of our existing databases we have, we will be only too happy to assist in connecting to your patient database.
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FAQs—SpectraView
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Wavelengths and Spectral Resolution: SpectraView
Overall, the spectral range of the system is 400–900 nm and is limited by the CCD chip and the optics. Efficiency in this range is not constant, and can be plotted as a graph that peaks at ~550–600 nm and decreases from this range at the edges. At 400 and 900 nm, efficiency decreases ~5-10% from the peak. Some data above 380 nm, but not below that wavelength, may be measured. Efficiency may be improved by measuring only the blue range, dividing the sample by two or three and measuring separately using a band filter. Such a measurement can improve the noise on the signal, but may take a longer time. The best spectral resolution is 5 nm @ 500 nm.
It is important to understand that the spectral resolution of the system, like every Fourier-based spectrometer, is constant in the energy domain (or frequency, or Wave number measured in cm-1), which means that the spectral resolution is not constant in the wavelength range. Resolution is better than four nm around 400 nm, and about 16 nm at 900 nm. The equation for the spectral resolution is approximately: dL = L^2 * 2*10^(-5) where dL is the resolution in nm and L is the wavelength in nm.
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What is the SpectraView System?
Our technology is capable of measuring a panel of immunoassays. The set of stains we use in our spectral karyotyping applications can be viewed as similar in many ways to immunoassays— detecting an image that has many colors, some of which are blended, and separating and measuring the quantity of each color at each point.
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FAQs—METScan
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Which objectives do you recommend for the MetScan?
ASI recommends that you first scan with a 10X objective and use a higher magnification for the second scan once the required metaphases are chosen.
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How long does it take to scan a slide for metaphases?
A full slide can be scanned between 3–7 mins. However, there are many factors affecting the timing, such as mitotic index and sample type.
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FAQs—SpotScan
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Is it possible to adapt SpotScan to different conditions?
Each ScanView application, METScan and SPOTScan, can be adapted to any condition. ASI ScanView has an intergrated training feature allowing the setup and parameters of the system to be adapted to different conditions.
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FAQ's—HiSKY
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What Type of C-mount should I use?
A 60X or 63X oil objective with a 1X C-mount is preferable as no magnification or reduction is required, but the user performing the analysis should decide the magnification or reduction according to the software. For example, if a 100x objective is used, a 0.7X or 0.63X C-mount is preferable because it provides a wider field of view (the CCD image covers a larger area of the sample due to the reduced magnification).
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Other FAQs
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What type of CCD does ASI use with their products?
All ASI acquisition products are supplied with a high-end 12-bit digital camera, with a 2/3” CCD and 1.3 million pixels
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I would like to use ASI images in my article or report. Who should I contact for this?
Please contact
This e-mail address is being protected from spambots. You need JavaScript enabled to view it
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I am living in a country that does not appear to be in your contact lists. Who should I contact for a quotation?
Please send all requests to
This e-mail address is being protected from spambots. You need JavaScript enabled to view it
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I already own a microscope. Can I use it with ASI products?
All ASI products can be adapted to any manual or automatic microscope that provides a camera and a C-mount.
For ASI Scanning products (ScanView) an Olympus BX61 or Zeiss Axioimager M1 or Z1 is required
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