LightCannon: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
No edit summary
Line 1: Line 1:
This page is a placeholder for how to build a light cannon for bacterial photography.  Expect the page to be complete before the end of January 2006. In the meantime, below are some raw emails that need processing [[User:Endy|Endy]] 15:56, 22 December 2005 (EST)
This page is a work-in-progress for how to build a "light cannon" for use in bacterial photography.  <br>
Please help make it better!
 
[[Image:ColiroidFSM.jpg]]
[[Image:ColiroidEllington.png]]
[[Image:LightCannon2.png]]


----
----

Revision as of 19:49, 22 December 2005

This page is a work-in-progress for how to build a "light cannon" for use in bacterial photography.
Please help make it better!


Anselm- The details on the filter are:


From Edmunds Optics description: FILTER INT 632NM 50.8MM SQ Catalog number: NT43-185 details on this website

http://www.edmundoptics.com/onlinecatalog/DisplayProduct.cfm?productid=1903


> >Hey Jeff, > > > >could you tell me the details about the bandpass filter you use? > >HWFM and CWL or related. (or even just supplier and #) > >Just want to make sure I'll hit the same optimum window with some that > >I'm buying. > > > >Thanks alot! > > > >-Anselm




our thermometer hole is ~1.5 inches.


At 03:53 PM 12/15/2005, you wrote: One last annoying question. the hole in the top of the incubator, how big is it? I'm looking at a few different ones and can't decide if the standard thermometer port is large enough.

thanks for answering all these questions!

-anselm



we've been using a 100W mercury vapor lamp, taken from a fluorescence microscope. with this light source we use a red bandpass filter (see the attached figure), that allows 660nm light to pass to the exclusion of far red light. we then place the image which is on a 35mm slide (the kind you would use for an old-style slide projector) below the filter, and focus the light through a double gauss lens. the whole apparatus sits outside of a 37C incubator, and the light image passes in.

one really nice feature that we have is an actuator to which 35mm slide is connected, which allows us to have pretty fine control over the focus.

we've also succesfully been using sodium "grow" lamps, which we don't filter usually.

as far as the agar preparation, it's still just 2-4% seaplaque agarose in LB+S-gal and Ferric Ammonium Citrate. We autoclave the media with everything in it before pouring, to get the solutes completely dissolved, then we keep it at 42 degrees, add the cells, mix and pour immediately.

we've actually thought about increasing signal to noise. one idea we had was to put a repressor of LacZ under the OmpF promoter. this will bring down total signal most likely but could also increase signal to noise. i'm sure there are lots of other good ways to do it as well...including mutating Cph8 and screening. good luck, let me know how it goes.

Jeff


At 09:11 PM 11/2/2005, you wrote: Chris said that you guys have been using a smaller setup to develop your bacterial photos, could you tell me what kind of light sources and filters you've been using? I'd like to start taking some myself to see if I can improve the signal/noise ratio by modifying the construct. Also, if you've changed the agar preparation at all, I'd love to hear what's been optimized. All details are great as I'd like to just order the things needed.

By the by, I'll finally be starting some tests on a possible blue-light sensor soon.

thanks a lot! kick ass regarding the paper! -Anselm