How can I photograph dim blue luciferase glow with a DSLR?
Asked 3/7/2012
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2 answers
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I’m trying to photograph luciferase bioluminescence, which appears blue around 480 nm. I can clearly see the glow by eye, but I haven’t been able to capture it with my Nikon D80 using 35mm and 50mm Nikon lenses. I’ve also tried removing UV/protective filters.
So far I’ve shot at 30 seconds, f/22, ISO 1600. Do I need a special lens or filter for this wavelength, or is this mainly an exposure and setup issue? What shooting setup and camera settings would give me the best chance of capturing the glow?
Originally by Photography Stack Exchange contributor. Source · Licensed CC BY-SA 4.0
Photography Stack Exchange contributor
14y ago
2 Answers
9
That wavelength is certainly within the spectrum you can capture with any lens, with or without filters. Digital sensors capture between 350-1000nm If it is glowing, then you'd want that to be your main light source. Any additional light you throw onto it is going to dilute the glow from your subject and make it harder to see.
What aperture are you using? Unless this is incredibly dim, I can't imagine 30" not capturing something unless you're using a very small aperture.
Are you doing this in a dark room with a dark background? What do you see in your 30" exposures?
Through trial and error, you could do a range of shutter speeds (and ISO) to try to hone in on a good exposure value. If you can see it with your eyes, you should be able to photograph it without special equipment.
Originally by user4191. Source · Licensed CC BY-SA 4.0
user4191
14y ago
0
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You do not need a special lens or filter for 480 nm blue light. That wavelength is well within what normal camera lenses and digital sensors can record.
The main issue is likely exposure and setup. At f/22 you are cutting the light dramatically, which is a poor choice for a very dim glowing subject. Try this instead:
- shoot in total darkness, with the luciferase as the only light source
- use a dark background so ambient light doesn’t wash out the glow
- focus first with room lights on, then switch to manual focus
- use manual exposure and bracket several shots
- open the aperture much wider than f/22
- vary shutter speed and ISO to find the best exposure
Any extra light will dilute or wash out the bioluminescence, so start with no other light at all.
If you can see the glow with your eyes, the camera should be able to capture it without special equipment, but it may require wider apertures, careful focusing, and trial-and-error with long exposures.
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