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http://totallab.com/spot-map » Accurate quantitation is an important part of your HCP analysis. In this video we show you how automatic spot outlines in SpotMap obtain an objective, accurate and quantitative analysis and how you can use 3D mode to validate the software you currently use.

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SpotMap: Accurate Quantitative Analysis for HCP Coverage 

Good image capture is critical to guarantee optimal performance of automated image analysis packages and generate reliable scientific data. This checklist defines some of the important technical factors required to generate digital images of a quality suitable for automated image analysis. Download our image capture guide here: http://totallab.com/cms/pdf/imagecaptureebook.pdf

Image Quality Hints and Tips- Checklist 

TotalLab software will analyse colour images. If the software loads a colour image it is automatically converted to grayscale for analysis. However, we recommend using greyscale images as you have the potential to get more data from your analysis. A colour image is made up of 3 different channels; Red, Green and Blue. So a 24-bit colour image has 8-bits per channel. When the image is converted to grayscale, it’s effectively changed to an 8-bit grayscale image with weighted conversion of the Red, Green and Blue channels. So a colour image, when imported to the software, will lose information and be converted in a way that may not be accurate for the image capture device used. Always scan directly to grayscale as the imaging device will then do the conversion from colour in the most accurate and sensitive manner. For more information download our image capture guide: http://totallab.com/cms/pdf/imagecaptureebook.pdf

Grey scale - Image Quality Hints and Tips

What is bit depth? The bit-depth of the image is important as it represents the number of available intensity levels that each pixel can take. Also referred to as colour depth or pixel depth, bit depth is the number of bits used to represent the grayscale (intensity level) of each pixel in an image. Greater bit depth allows a greater range of shades of grey to be represented by a pixel. Our recommendation: As a rule, the more levels of grey represented in an image, the better the ability to differentiate low abundance spots from the background, and the greater the quantitative accuracy. For more information download our image capture guide: http://totallab.com/cms/pdf/imagecaptureebook.pdf

Bit Depth - Image Quality Hints and Tips

The best resolution at which to capture your images will be dependent on the size of the image you are capturing. Image (or spatial) resolution relates to the number of pixels displayed per unit length of a digital image and is often measured in dpi (dots per inch) or in microns (the size of the area each pixel represents). Images with a higher spatial resolution have a greater number of pixels and have more image detail than that those of lower spatial resolution. Our recommendation: Try to scan at the best resolution for your images. In most situations, 300 dpi or 100 microns will provide an image that is large enough for accurate analysis and small enough for efficient processing. For more information, download our image capture guide: http://totallab.com/cms/pdf/imagecaptureebook.pdf

Image Resolution - Image Quality Hints and Tips 

It is important to avoid saturation effects. Saturation occurs when grey levels exceed the maximum available. When a spot becomes saturated, any differences in high pixel intensities cannot be resolved, and the spot appears truncated when viewed in 3D. Why can saturation be a problem? The outline detection is not optimised for the unnatural shape of these spots, since the peaks are effectively ‘missing’ in the scanned image, and no reliable quantitation can be done with them. Saturation can cause issues in spot detection. It also prevents accurate quantitation of the spots as the true peak height cannot be known. To find out more, download our image capture guide: http://totallab.com/cms/pdf/imagecaptureebook.pdf

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