[help] about plate reader wavelength
Posted 19 January 2011 - 09:02 PM
I was planning to run Glutathione assay kit for my project using following kit.
Unfortunately, I found out that 405nm filter on my plate reader is faulty (Lamp energy too low) and cannot be used.
But I cannot wait to get it repaired. Their recommended range is 405-414nm.
Which other wavelength can I try instead of 405? I have options of 340, 450, 490, 540, 595, 750nm to choose from.
What possible things can happen if I use other filter other than recommended nm? But I'm willing to take the risk.
Any advice is appreciated.
Posted 19 January 2011 - 10:34 PM
Posted 20 January 2011 - 04:13 PM
Posted 23 January 2011 - 02:06 PM
But in addition to the recommendations above; you ought to be able to continue with the 405nm so long as you have an appropriate standard curve albeit with reduced readings and sensitivity.
Posted 26 January 2011 - 07:29 PM
I also contacted technical support of the kit and replied to try at 450nm(closet to 414), though with reduced sensitivity. Anyway, it worked more or less.
@DRT, Sorry, I don't get about difference between light source vs filter.
In that machine, only 405nm doesn't work. Other filters (450, 595, 750) just work fine.
Anyway, I contact the instrument vendor and they said; need to change the lamp.
Posted 27 January 2011 - 03:17 PM
Posted 27 January 2011 - 11:59 PM
The lamp energy is a measure of the amount or "strength" of the light coming out of the light bulb in the machine. What you perceive as white light is actually made up of lots of different wavelengths, some stronger and some weaker in output (see: Sodium bulb emission spectrum. As the bulb gets older, the bulb produces less light, some of which happens to be the at the 405 nm wavelength. The machine calibrates the amount of light it receives from the bulb and uses this as a basis for measurement... if too little light is received, it can't calibrate itself and give you an error.
Thanks for concise explanation.