Macroscope

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Line 41: Line 41:
   * YFP: HQ500/20x 25mm
   * YFP: HQ500/20x 25mm
   * mCherry: Chroma [http://www.chroma.com/products/part/1135 ET572/35x 25mm]
   * mCherry: Chroma [http://www.chroma.com/products/part/1135 ET572/35x 25mm]
 +
  * BFP : Chroma [http://www.chroma.com/product/individual-filters-mirrors/widefield-microscopy/D395_x_40x-EX]
=== Emission filters and automated filter wheel ===
=== Emission filters and automated filter wheel ===
* Emission filters. Chroma clearance bin as possible (50mm filters = $$)
* Emission filters. Chroma clearance bin as possible (50mm filters = $$)
Line 48: Line 49:
   * YFP: Shares GFP emission filter
   * YFP: Shares GFP emission filter
   * mCherry: HQ645/75m 50mm
   * mCherry: HQ645/75m 50mm
 +
  * BFP: Chroma 460/40 55mm (oops, should have been 50.  Works anyway, not perfectly.)
* [http://www.optcorp.com/product.aspx?pid=354-1630-1632-13255 Starlight Xpress USB 5 Position 2" Filter Wheel - 72mm Male Input/ Output]. Make sure here to get a filter wheel insert that is for unthreaded filters!  
* [http://www.optcorp.com/product.aspx?pid=354-1630-1632-13255 Starlight Xpress USB 5 Position 2" Filter Wheel - 72mm Male Input/ Output]. Make sure here to get a filter wheel insert that is for unthreaded filters!  
=== control box ===
=== control box ===

Revision as of 19:23, 12 February 2013

In the beginning, there was Disco!

coming soon ...


Contents

Macroscope:

what

why

capabilites (current and potential)

To build:

parts list and sources

basic structure

custom parts (cam and filter mounts)

  • Filter wheel mount. McMaster-Carr part *** machined to this spec ^^^. This part can also be ordered directly from emachine shop. Requires mating to the adapter that comes with the filter wheel (picture links).
  • Camera mount. McMaster-Carr part *** drilled with 1/4" bit to allow mating to camera tripod mount.
  • LED filter coupler. 1 per LED. Available through eMachine Shop. design link.
  • LED mount and heat sink. 1 per LED. Available through eMachine Shop. design link.

camera

  • Canon T3i, with Canon EF-S 60mm f/2.8 Macro USM Lens, and Canon ACK-E8 AC Adapter Kit for Rebel T2i
  • Canon SDK can be requested here.

LED illumination packages. filters and leds - chroma...clearance...mention gel filters...

  * Wide dispersion
  * Medium dispersion 
  * Narrow dispersion used for fluo illumination.
  * CFP
  * GFP
  * YFP
  * mCherry
  * White
  • Excitation filters. Chroma clearance bin as possible.
  * CFP: Chroma D436/20x 25mm
  * GFP: Chroma S470/40x 25mm
  * YFP: HQ500/20x 25mm
  * mCherry: Chroma ET572/35x 25mm
  * BFP : Chroma [1]

Emission filters and automated filter wheel

  • Emission filters. Chroma clearance bin as possible (50mm filters = $$)
  * CFP: Chroma D480/40m-50mmr
  * GFP: Chroma D530/20  50mm
  * YFP: Shares GFP emission filter
  * mCherry: HQ645/75m 50mm
  * BFP: Chroma 460/40 55mm (oops, should have been 50.  Works anyway, not perfectly.)

control box

plate registration

Software (link to canon SDK req; starlight express drivers, switch&sense)

etc

Thoughts:

notes

  • Fluorescence shading correction. Divide images by a flat field standard
  • Filters. Interference filters are far and away the most costly part of this device, but have an enormous impact on signal to noise ratio. A very inexpensive but lower S/N filter option is to use theatric gel filters, available with transmission spectra from RoscoLUX, ... (ref worm breeders). A still cheaper option is to see how much fluorescent signal can be extracted using just the RGB filtered channels of the cameras sensor, along
  • Often, grabbing a single channel of the output fluorescent image (eg, Blue for CFP emission) further cuts some background and may improve the signal for certain fluors.

improvements

  • Better darkfield illumination
  • 3D imaging of fluorescence in transparent volumes using sheet illumination (laser, cylindrical lens, )
  • Hyperspectral imaging to allow fluorescent compound identification and spectral unmixing of combinations of FPs, and for transmission spectroscopy...(ref)

Similar Tech


  • sample images?


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