Suggestions - Nice Western Blot Image from Office Scanner? - (Jun/18/2013 )
We don't have a laboratory imager so I've always had to scan my western blots (colormetric) using a standard office scanner. It has always worked o.k. but I don't get great or crisp images,especially when the signal isn't strong to began with. I can see it looking at it but the scan makes it so I have to squint.
Does anyone have any suggestions of how to get nice western blot images using an office scanner?
Is it possible to change the exposure of the scan? I am sure that you can increase the file size to produce a sharper image. What are you saving your picture as (.tif, .jpeg, etc.)? Does your lab or a neighboring lab possess filters for imaging EtBr, Coomassie or other staining techniques?
We've played with some of the options on the scanner (sharpness, resolution, etc) but it has been difficult to pick up the clean, somewhat lighter bands without them looking pixelated and fake. We save the images as .jpeg's. Would what we save as have an effect? We don't have filters or access to them but I'm not sure I understand how they would apply to the office scanner. We've also tried laying bright white paper or filter paper behind the blot but that appears to make things worse...
I was hoping you would have some kind of imager for capturing gels. Those usually have a couple of different filter settings and one may be able to create a longer exposure to pick up your fainter bands.
Yes, what you save the scanned image as can have an effect. I would recommend .tif or .png. I have attached a link for reference.
I would continue to play with the scanner settings. Try to scan at the highest resolution/file size as you can. From there you could open your picture in an imaging software and bring out the lighter bands.
http://www.scantips.com/basics09.html
I use office scanner for scanning my blots. I agree with JerryShelly1, saving as tiff is definitely better and increasing dpi also seems to help. Are you using photo scanner?
as jerryshelly1 and iceage said, use highest possible mechanical resolution (not interpolated resolution) and save as tiff. also scan at the greatest bit depth (most colors or most shades of gray).
if you still can't get a good scan then you may want to try using a (high megapixel) camera (again, use highest resolution and bit depth). save as a raw file then convert to tiff for manipulation and presentation.
Thank you all for your replies and for the scan tip reference link. I will try switching to .tiff and scanning at the greatest depth and see how that goes. I wish a good quality imager didn't cost so much $$!
It wouldn't be a bad idea to consider mdfenko's advice. Make a cardboard cone similiar to the one provided in the link below. With this you could use a high megapixel camera to take the photos. With most camera's you could adjust the exposure to obtain all bands.
http://walkercameras.com/pics/ilford/pinhole-cones.jpg