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select from the pulldown. A histogram representation of your image will appear in the large box at the
bottom of the System window, to help you decide if your image is bright enough but not too bright.
To acquire an image press the “Snap” button. Then save the image using the “Save” or
“Save As” button at the bottom of the Acquired image window. The default file format for saving single
images is a .tif. Other file type options are available using the File Save As feature from the main
ImageJ toolbar menu. If you want to keep a snapped image, you must save it right away, otherwise it
will be overwritten by live preview data or another snap image acquisition. If you want to change cubes
between captures, you have to manually slide the filter slider between captures. If you want color channel
images and DIC images, use the foil “shutter” to block the brightfield light when you are taking color
images, it’s easier and more energy efficient than turning the bulb on and off with the power switch. The
ROI button will allow you to capture only a portion of the image. Use the box tool from the main ImageJ
toolbar to draw a box around a portion of your image, then hit the button with the blue box under ROI.
Only that portion of the image will be captured, saving space in memory. To return to the full chip, hit
the button with the four green arrows. If your image becomes too bright, too dim, or just plain funny
looking after you draw the ROI, try going back to full chip, then make sure that the “auto-stretch” feature
is turned off by unchecking the box. Hit the “auto” button to re-scale your image and then try drawing
your ROI again. A feature for storing and re-visiting multiple ROIs is built-in to the software, but
unfortunately since the software has no control of the microscope stage it is not possible to use this
feature to collect multiple data points from a
single dish/slide at this time.
Also, because the software
cannot control the microscope’s filters or
objective position, automated multi-channel
(“Channel group”) or multi-z plane (“Slices
(z)”) imaging is not possible. However, time
laspe imaging of a single channel (one color or
DIC) is possible. To collect a time series, hit
the Acquisition button on the System window.
You will get this Multi-dimensional Acquisition
window. Once you have set your exposure time
in the System window, use the “Frames (time)”
box in the Multi-dimensional Acquisition
window to set up your time lapse. Set the
acquisition order to time-lapse, and press
“Acquire!” to capture the images. Once the
capture is finished, you can then save your
images using either the “Save As” button from
this window (if you want a series of .tif files in
a folder created with the filename that you
supply) or the File Save As feature from the
main ImageJ taskbar (if you want another filetype, such as a .tif stack .stk). If you want to write your data
to disk as you go (nice for time lapses lasting several hours) you can set up a routine to write your images
to disk as soon as they are captured. This could save you from data loss if the computer and/or
microscope were to lose power at some point during your experiment.
(122 strony)
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