Appendix G/F13: Difference between revisions

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== APPENDIX G: Use of the Canon A620 and A630 Digital Cameras''' ==
== '''APPENDIX G.  Instructions for Capturing Digital Images of Nucleic Acid Gels Stained with SYBR Safe Using the BioRad Imaging System''' ==
Focus your microscope slide image.<br>
'''Instructions for Taking a DNA gel image stained with Sybr Safe using the BioRad ChemiDoc MP System with Image Lab Software'''
Record the total microscope magnification in your laboratory notebook.<br>
 
IMPORTANT:
We own several Canon digital cameras:  A620 models with a resolution of 7MP (megapixels) and A630 models with a resolution of 8MP. The A620 and A630 models are essentially the same camera.  
Ethidium Bromide stained gels may NOT be imaged in this equipment.
Our strategy will be to capture images at the highest resolution (7 or 8 MP) of the camera, transfer them to the computer and use Photoshop to edit.
Remove gloves and wash hands BEFORE using the computer. DO NOT contaminate the computer.
#Pull out the viewfinder and flip it so you can see the screen on the back of the camera.
The XcitaBLue Conversion Screen should be kept covering the UV transilluminator and only removed TEMPORARILY when using fluorphores other than Sybr Safe or SybrGreen. Please return the Xcita Blue Screen to the UV transillumintor if you remove it.  
#To take a photo, make sure the switch on the back of the camera is set to the red-camera-icon position (live image mode).
 
#Set the camera to auto on the wheel at the top of the camera near the on/off button.
'''Quick and Easy Protocol for photographing Sybr Safe DNA gels:'''<BR>
#Turn off the flash by pushing the function wheel on the back of the camera under the lightning (flash) symbol. If the flash is off the flash symbol will have a line through it on the screen.  
1)Make sure the Power Button on the right front of the imager shows a green light. If not press it until the green light comes on and wait 5-10 min for warm up.  
#Push the zoom all the way to the left so that you've zoomed out all the way.
 
#Place the camera lens against the microscope ocular lens containing the ocular micrometer.
2) Open the UV transilluminator drawer on the lower front of the imager and make sure that the XcitaBlue Conversion Screen is in place. If not find it and position it covering the glass. Clean it with water and a paper towel if necessary.
#Move the camera lens around the microscope eyepiece to find the image. This takes a little practice and patience. The camera lens should not touch the microscope eyepiece.  The image field is circular.
 
#Rest your left hand on the side of the microscope eyepiece and cup your fingers around the lens of the camera and the microscope.  This will stabilize the camera and facilitate obtaining the best image.  The camera lens should be about a quarter of an inch or so from the ocular lens.
3) Position your gel in the center of the glass.
#The camera will automatically focus the image. When you have obtained an evenly illuminated circular image field, gently press the exposure button at the top of the camera to take the picture.
 
#To transfer captured images to the computer, slide the switch on the back of the camera from the red-camera-icon position (live image mode) to the blue-arrowhead-icon position (captured-image review mode).
4) Close the drawer, remove gloves, and wash your hands before using the computer to the left of the imager.  
#Plug USB cable into computer keyboard port located on back of keyboard, then into camera port. The iPhoto window opens up. Check box to delete image from camera.  
 
#Click Import to download your image into the iPhoto Library folder.  Confirm that the “Delete originals” button is selected.
5) Open the ImageLab 4.0.1 software by double clicking on the icon on the computer desktop
#In the File menu, select Export...  The Export Photos window opens.  Select “Full-size images” and “Original” format.  Click Export button.
 
#A window shade drops down in the Export Photos window.  Rename your file in the Save As text box and choose Desktop as your destination.  Click OK.  Close the iPhoto window.
6) Find and open the Protocol called SybrSafe under Recent Protocols or from the Open Menu. Double click to open it.
#Launch Adobe Photoshop.  In the File menu, select Open…  The “Open” window appears.  Find and select your image, then click Open.
 
#In the Embedded Profile Mismatch window, select “Use the embedded profile…” and click OK.  Your image opens.
7) Click Position Gel (yellow button) to check the position of your gel.
#Select the “Rectangular Marquee Tool” by typing the letter M on the keyboard.  Drag cursor to select desired portion of your circular image, including a segment of the ocular micrometer. This yields a conventional rectangular image suitable for use in a paper.
 
#In the Image menu, select Crop.  The selected portion of your picture appears on the desktop.
8) Click Run Protocol (green button) to take the photo.
#In the Image menu, select Image Size...  The “Image Size” window appears.  Make sure the Constrain Proportions box is checked.  In the Document Size portion of the window, decrease the larger of the two image dimensions to 4 inches.  Click OK.  
 
#In the File menu select Save As… and rename file.  Use JPEG as the image Format.  Click Save.  JPEG Options window opens.  Set Quality to Maximum (10).  Click OK.  This will create a small file of about 150–300KB in size.  Delete the original large (3MB) image file.  
9) To Save your image (if you are not going to analyze it quantitatively), find or make a folder for your work in Documents (NOT on the Desktop). go to File---Export---Export for Publication (use the defaults, e.g. 300dpi). You will see a pop up message reminding you that this option is not optimized for analysis---it doesn’t matter—Click OK.  Check the Location where your image will be saved, the FILE Name, and use the drop down menu to SAVE AS TYPE tiff or jpg. Click Save.
#Email the images to yourself and to your partner(s). Check with your instructor to if she/he would like them uploaded to the lab Sakai site.
 
10) Close the Image Lab Software
11) Don gloves, remove your gel, clean the glass with water and a papertowel.
12) The computer AND the ChemiDoc Imager should remain ON. DO NOT Turn OFF the power or shut down or log off the computer.

Latest revision as of 05:11, 21 October 2013

Wellesley College

BISC110/112- Introduction to Cell Biology Lab- Fall 2013

   

APPENDIX G. Instructions for Capturing Digital Images of Nucleic Acid Gels Stained with SYBR Safe Using the BioRad Imaging System

Instructions for Taking a DNA gel image stained with Sybr Safe using the BioRad ChemiDoc MP System with Image Lab Software

IMPORTANT: Ethidium Bromide stained gels may NOT be imaged in this equipment. Remove gloves and wash hands BEFORE using the computer. DO NOT contaminate the computer. The XcitaBLue Conversion Screen should be kept covering the UV transilluminator and only removed TEMPORARILY when using fluorphores other than Sybr Safe or SybrGreen. Please return the Xcita Blue Screen to the UV transillumintor if you remove it.

Quick and Easy Protocol for photographing Sybr Safe DNA gels:
1)Make sure the Power Button on the right front of the imager shows a green light. If not press it until the green light comes on and wait 5-10 min for warm up.

2) Open the UV transilluminator drawer on the lower front of the imager and make sure that the XcitaBlue Conversion Screen is in place. If not find it and position it covering the glass. Clean it with water and a paper towel if necessary.

3) Position your gel in the center of the glass.

4) Close the drawer, remove gloves, and wash your hands before using the computer to the left of the imager.

5) Open the ImageLab 4.0.1 software by double clicking on the icon on the computer desktop

6) Find and open the Protocol called SybrSafe under Recent Protocols or from the Open Menu. Double click to open it.

7) Click Position Gel (yellow button) to check the position of your gel.

8) Click Run Protocol (green button) to take the photo.

9) To Save your image (if you are not going to analyze it quantitatively), find or make a folder for your work in Documents (NOT on the Desktop). go to File---Export---Export for Publication (use the defaults, e.g. 300dpi). You will see a pop up message reminding you that this option is not optimized for analysis---it doesn’t matter—Click OK. Check the Location where your image will be saved, the FILE Name, and use the drop down menu to SAVE AS TYPE tiff or jpg. Click Save.

10) Close the Image Lab Software 11) Don gloves, remove your gel, clean the glass with water and a papertowel. 12) The computer AND the ChemiDoc Imager should remain ON. DO NOT Turn OFF the power or shut down or log off the computer.

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