Titanium Phosphate Glass Microspheres as Microcarriers for In Vitro Bone Cell Tissue Engineering - Biointerfaces: Where Material Meets Biology - page 118

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Figure 5.3 Screenshots of multi-dimensional acquisition and Snap/Live windows in

mManager software.

5. Carry out the following steps to obtain microsphere diameter values
from the stored time lapse images using ImageJ version 1.47.
i. Open the time lapse images: Go to File | Import | Image Sequence.
Go to the folder containing the images. Click on the first image
from your sequence and press Open. Press OK on the window that
appears.
ii. Binarise the images: Go to Process | Binary | Make Binary and press
OK on the window that appears.
iii. Select a rectangular area around a sphere: Press on the rectangle
icon and then draw a rectangle on the image wherever required by
dragging the cursor (which looks like a ' þ ' on the image) across
the image with the left mouse button pressed. The position of the
rectangle can be adjusted by keeping the cursor anywhere inside
the rectangle (the ' þ ' sign changes to a normal arrow) and drag-
ging it over the image.
iv. Exclude all objects other than the ones inside the rectangular area:Go
to File | Edit | Clear Outside, and press OK on the window that
appears. Remove the rectangular area by clicking anywhere on the
image.

.

TIP: Before excluding all other spheres from each image, make sure
that the bead of interest stays fully within the rectangle throughout
all the images by moving the horizontal scroll bar from beginning to
end. It is alright if parts of other spheres are visible inside the rect-
angular area since the software will exclude them during the analysis.