Transcript Powerpoint - University of Northern Colorado

An Inexpensive Gel
Documentation
System
Tony Schountz, PhD
School of Biological Sciences
University of Northern Colorado
[email protected]
http://www.unco.edu/schountz/gel/
Revision 3.2
16 Oct 2006
Commercially-available gel documentation
systems cost US$2,000 or more, but usually have
software for sophisticated gel analysis.
This document describes the assembly of a gel
documentation system (image capture) for about
US$250. It requires basic skills in power tool use
to build a light box for a commercially-available
digital camera, the Canon PowerShot A520.
The idea for this system was adapted from
one used by the Hirsh Lab
http://www.virginia.edu/biology/Fac/hirsh_gel/
gel/
Although this system is working for me, I offer
no
guarantees that it will work for you!
Camera Equipment
(Can be purchased from B&H Photo*)
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Canon PowerShot A520 (B&H #CAPSA520) - $170
LA-DC52F 52 mm filter adapter for A520 (B&H #CALADC52F) $18
52 mm UV filter (B&H #BWUV52) - $20
52 mm infracolor 099 orange filter (B&H #BW09952) - $20
1/4” Tripod screw (B&H #LI861568 or Home Depot) - $4.50
ACK-800 AC Adapter Kit (B&H #CAACK800) - $40 (optional eliminates the need for camera batteries)
B&H Photo http://www.bhphoto.com/
*I have no financial interest in B&H
Photo
Canon PowerShot A520
• PowerShot A520 Body (4 megapixels)
• MultiMediaCard memory card (included
Needed items included with the camera
with most cameras)*
• 2 AA Alkaline Batteries
• Canon Digital Camera Solution CDROM
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USB Interface Cable
*There is no need to purchase a larger memory card because
the images are stored on your hard drive, not the camera’s
memory card
Camera Note
Although this document discusses the use of the PowerShot
A520, other Canon cameras should work, so long as there is
a filter adapter for the camera. So far, we have used three
Canon cameras with success, the PowerShots A520, A95
and A70. These are no longer in production, but Canon
offers others with similar prices and features. For additional
information, see Canon’s web site:
http://www.usa.canon.com/html/canonindex.html
Software
• Canon Digital Camera Solution Disk
(included with the Canon camera)
• ImageJ - http://rsb.info.nih.gov/ij/
• Gel-converter.class plugin for ImageJ,
available at the Hirsh lab web site
• Optional: iPhoto 5 (Mac users only)
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Hardware
USB-equipped Mac (OS X) or Windows PC
Transilluminator
Five sided box (1/2” width boards) with 2.25” hole in the
top to accommodate the camera filter adapter
1/2” Weather stripping
L-bracket with 1/4” hole for tripod screw (e.g., Home
Depot)
1/4” nut for the tripod screw (to secure it to the Lbracket)
Handles for the sides of the box
#6 x 3/4” screws for holding the box together; #5 x 5/8”
screws for attaching the L-bracket
Tools
• Table, circular, or jig saw
• Straight edge for cutting guide
• Drill and bits
• Screw driver
• Optional: Router with flush-trim bit
• Optional: Random orbit sander/220+
paper
Box Construction
Here’s how I made my camera box1:
1/2” Medium-density fiberboard (MDF; Home Depot, Lowes,
etc.).
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10”d x 11”w x 10”h. I cut four 10”x10” boards for the sides
and a 10”x11” top. You’ll need to custom-design a box to fit
your transilluminator. The height should be enough so that
the camera can see all of the glass on the transilluminator. I
used a router with a flush-trim bit to get nice smooth edges.
Held together with wood glue (probably not necessary) and
countersunk #6 x 3/4” wood screws. Pre-drill the holes for
the screws. Use a hand-held screwdriver, and not a powered
screwdriver, to tighten screws in MDF. Do not over-tighten!
2.25” hole for filter adapter was cut with a hole saw (about
$11 from Home Depot) for power drill.
Lightly sand the outside surfaces. Prime, then spray-paint flat
black inside and outside. This is important because it
eliminates reflections and flare inside the box.
Handles attached with screws only.
1/2” weather stripping on bottom edges to protect the
transilluminator glass.
Note: If you don’t have access to a power
saw or are uncomfortable using one, most
home centers (such as Home Depot or
L-bracket screw hole drilled out to 1/4” to accommodate
Lowes) will custom cut MDF to size for a
tripod screw. Important: Mount the L-bracket to the camera
before marking the drill holes for mounting the bracket to the small price. Usually, the first two cuts are
box. This is necessary because the filter adapter protrudes
free, the rest are about $1 each. The
into the box. Use appropriate screws for the L-bracket (I
accuracy of these cuts are adequate for box
used #5 x 5/8” wood screws).
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System Assembly
Attach filter/lens adapter to
camera
Attach UV filter to adapter
Attach orange filter to UV filter
Thread tripod screw through Lbracket, then onto the camera.
Visually align the camera so that
it is perpendicular to the
transilluminator glass.
Position the box on the
transilluminator
Note the USB and power adapter cables are plugged into the
camera.
The Maxell AC 3000 power adapter (Circuit City),
pictured above, is plugged into the outlet (red box) and
connected to a PowerShot A70. The PowerShot A520 has a 3.2
V requirement. The AC 3000 has a voltage setting (3.3 V) close
to the A520’s requirement. The Canon ACK-800 adapter is about
$7 more than the AC 3000. Other Canon cameras have different
voltage requirements, so exercise caution when setting the
voltage on the adapter.
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System Setup
Install the Canon software
Install the ImageJ software
Install the gel_converter.class plugin into ImageJ’s plugin folder
Connect the camera to a USB port
Make sure the camera is set to Playback Mode using the Mode
Switch
If you’re using Mac OS X, open the application Image Capture.
In the Preferences, set the camera connection option to open
the CameraWindow application found inside the Canon
directory. Quit Image Capture. From now on, anytime the
camera is turned on, CameraWindow should automatically
launch.
Turn the camera on; CameraWindow should launch
CameraWindow
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Canon software utility that
controls the camera from the
computer
After it opens, click on Remote
Shooting
Click on the icon for Starts
Remote Shooting
You’ll be prompted for where
the image files are to be saved.
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Remote
Shooting
Size and
Resolution
Rotation
Setting
The RemoteCapture Task application
is used for capturing gel images
You should see a live video of what’s
on the gel box
Adjust the gel’s position so that it’s
roughly centered in the video window
Zoom using the zoom slider
Set all the parameters for Size and
Resolution (set to SS for smallsuperfine here), Rotation Setting, and
the Field angle/flash settings as
shown in the picture
Zoom
Slider
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Remote
Shooting
Adjust other settings in the Shooting
settings tab. You may want to adjust
some of these settings to suit your
needs. We set Photo Effect to Black &
White and sometimes switch AE Mode
to Manual if we have dim bands.
When the gel is properly framed, click
the Release button (encircled)
The image will be saved in the folder
you selected at application launch
Example Gel Capture
Genomic DNA digested with PvuI
This image was reduced to 640x480 from the original
1024x768, but otherwise unaltered.
Gel Editing
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Open the gel file in
ImageJ
Crop and rotate as
desired
Run the Gel Converter
routine from the Plugins
pull-down menu
Scale the image as
necessary
The Gel Converter routine
produces an inverted
grayscale image of the
gel
Note: By inverting the grayscale, less ink or toner will
be used when the image is printed.
iPhoto
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You can import gel files into iPhoto for easier management.
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Choose File > New Album
Choose File > Add to Library...
Select files to import
Drag and drop the image files into the newly created album
iPhoto will provide you with exposure information as well as date and
time the photo was taken.
iPhoto can also convert the color images to black and white, and it
performs basic image manipulations, such as cropping and contrast.
Notes
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Be sure to get the correct lens/filter adapter for the Canon camera that
you purchase.
The orange 099 filter is not a narrow bandwidth filter. Narrow
bandwidth filters are substantially more expensive ($200+), but
probably give a little better image.
Our system is used with an iMac G5 (OS X 10.4.3) and Canon
software CD version 19.
Comments, suggestions, questions and complaints can be emailed to
me. I’ll do my best to answer questions and address complaints.
If you end up using a system based upon this presentation, please let
me know so I can score points with my department director!
Email: [email protected]