These images were obtained by fluorescence imaging of fibroblast cells. The actin and mitochondria are each stained in such a way that they fluoresce at different excitation wavelengths. Green light makes the microtubules fluoresce red, while blue light makes the actin fluoresce green. The intensity values for each of these emmissions are collected separately. These separate images are then adjusted so that they display a full range of intensity values and are composited into a single RGB image with the mitochondria in the red channel and actin in the green. Post-processing was accomplished with ImageJ.
To estimate scales at both 20x and 100x magnification, we took three measurements of the calibration slide at each magnification. From these three measurements we calculated average pixel to length ratio with ImageJ using the length information provided by the manufacturers of the slide.
Using the scale information previously calculated, we superimposed scale bars on top of the images using ImageJ.
We imaged the spatial calibration slide at group 5 element 2. We estimated that 1440 pixels equals 1mm.
The microscope digital image capture device does not capture colors correctly. Either there are settings we are not familiar with or the device needs to be fixed/calibrated. We attempted increasing the brightness of the blue channel in our post-processing to achieve a better image. Although this does not fix the problem completely, it made an improvement. Regardless, no color information is neccessary to actually study the spirulina.
Based on further eyeballing and discussion we were able
to estimate the following:
- ~65 turns of the helix within half the field of view.
- The thickness was approximately a quarter of the pitch
- The outer diameter was roughly twice the thickness
This enabled us to come up with the dimensions shown on the illustration below and compare it to more accurate dimensions acquired by imaging with a microscope.
By eyeballing we underestimated the dimensions of the spirulina by about a factor of two.
