FAQS - Spectral Imaging

	FAQs
	Faq's - BandView®

		When I captured the Metaphase, there were some chromosomes not captured. How do I add this?
		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.

		Can I annotate my images
		Annotation of any image can be done in Metaphase or karyotype mode
	Faq's - Multi-Species

		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.

		I work with chromosomes from rare species. Can I use BandView for this?
		With the addition of the MultiSpecies module, all species can be karyotyped
	Faq's - FISHView

		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.

		Which Fluorochrome filters do I need for my application?
		The Fluorence filters in the microscope are used to select the appropriate signal spectral which are emitted from your specimen. Before we are able to advise you on which filters to purchase, we need to know about the application and which probe kits you wish to use.
	Faq's - SkyPaint® Probes

		What is the content of vials no.2-4?
		Vial 2 contains a blocking solution to reduce the background, this step is an optional step and can be omitted from the process. Vial 3 and 4 contain antibodies conjugated to Cy5 and Cy5.5.

		What can be done to improve my results if the Cy dyes are weak?
		You can repeat the detection step. Remove carefully the coverslip and wash the slides for 5 minutes in 4XSSC/).1%Tween 20. Continue with step 5 in the detection step.

		For how long can I keep the kit?
		Under proper condition (at 4°C, vial no. 1 can be stored at –20°C) the kit is stable for 6 months, after this period the antibodies (vials 2-4) should be replaced. The SkyPaint-DNA kit (without vials 3 and 4) and the CAD (concentrated antibodies) kit is stable for 9 months under proper conditions (stored at -20°C)

		Can I use other types of pretreatment?
		Yes, Proteinaze K can be used. In our hand a brief incubation in Trypsin (1/2-1/5 of the concentration that is used for G-banding) is also very useful to remove residual cytoplasm.

		Can I use a rapid wash without Formamide?
		Yes, you can wash your SKY slides in 0.5XSSC for 5 min at 72C and then for 2 min at 4XSSC/0.1%Tween20.

		Can I hybridize my slides for only 16-24 hours?
		Yes, you can hybridize your SKY slide for only 16-24 hours and usually get very good results.

		Can I denature the slide and the probe together (codenaturation) ?
		Yes, you can use codenaturation for the slide and the probe. In this case no pretreatment : Protocol for codenaturation Put 10ml from the probe on the slides, cover with a cover glass and close with rubber cement. Put on a hotplate at 75C for 1.5 minute Transfer immediately to the incubator at 37C for 16-24 hours Continue with the washing step
	Faq's - Case Data Manager

		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 the existing databases we have, then we will only be too happy to assist in connecting to your patient database.
	Faq's - SpectraView

		Wavelengths and Spectral Resolution SpectraView
		The spectral range of the system is limited mainly by the CCD chip (also by the optics) and in overall the range is 400-900 nm. The efficiency in this range is not constant, and it can be plotted as a graph that peaks around 550-600 nm and goes down from this range to the edges. At 400 and 900 nm, it will go down to about 5-10% from the peak. While it may measure some data above 380 nm, it will not measure any data below that wavelength. Also, it is possible to improve the measurement by measuring only the blue range by dividing the measurement to say 2-3, and measuring it separately (by using a band filter). Such a measurement can improve the signal to noise, but it may take a longer time. In short, 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, and it is better than 4 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.

		What is the SpectraView System?
		Our technology is capable of measuring a panel of immunoassays. In many ways one can look at the stains that are used by us for the applications in spectral karyotyping as a set that is similar to immunoassays. It is different from the biology point of view, but the system is doing the same - it is detecting an image that has many colors, some of them are mixed together in the same geographical areas, and it knows how to separate it and give the quantity of each color at every point.
	Faq's - METScan

		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 required metaphases are chosen.

		How long does it take to scan a slide for metaphases
		A full slide can be scanned anywhere between 3-7 mins. However there are many factors effecting the timing such as mitotic index and sample type.
	Faq's - SPOTScan

		Is it possible to adapt SpotScan to different conditions?
		Each of the ScanView applications, METScan and SPOTScan can be adapted to any condition. ASI ScanView has an intergrated training feature whereby using the setup and parameters the system can be adapted to different conditions.
	Faq's - HiSKY

		What Type of C-mount should I use?
		Recommended using 60X objective with 1 X c-mount can work when the user performing the analysis only by the software Depending on the magnification of the objectives the reduction or magnification of the c-mount should be decided. e.g. if you are using a 100x objective a 0.7x (or 0.63x) c-mount is preferable because it gives you a wider field of view (the CCD image covers a larger area of your sample due to the reduced magnification). If the customer is using a 63x (or 60x) objective a 1x c-mount (no reduction or magnification) is preferable.
	Other Faqs

		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

		I would like to use ASI images in my article or report. Who should I contact for this?
		Please contact sales@spectral-imaging.com

		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 sales@spectral-imaging.com

		I already own a certain microscope. Can I use this 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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