What causes the broadening of diffraction spots in the TEM at parallel illumination and high CL?

As the title says, my question is about the diffraction spot intensities. I quantify the diffraction spot intensities for my analysis but accidentally I get the diffraction images in one series at 480mm instead of 300mm. I am comparing the intensities with different series of images, does it affect my work? 

I am analyzing amphibole samples with TEM and I don't have equipment yet to analyze the diffraction pattern (camera, software...). What do you think I shoud buy ?

Our institute require a new tem camera for the in-situ video recording for JEOL JEM 2100F TEM. I would like to ask about the camera specifications and their importance on imaging. 

First of all, the field of view and sensor size relation. one brand camera has 8.8*6.6 mm sensor and 25.5*19.2 mm field of view however other brand provides 15*15 mm field of view and same sensor area. what would be the effect of these parameters ?

2- what is the advantage or disadvantage of fiber optic or lens coupled CCD?

and also 3:1 and 1:1 tapering

3-most important one is the resolution. what will be the effect of 2.8 or 4megapixel resoultion on imaging and video recording. Since our microscope is not an atomic resolution microscope 2.8 megapixel camera could cause any significant disadvantage against 4 megapixel one. We are more focused on 2.8 mpx camera because its video recording speed is much higher both at full resolution (55 fps) (no info for 4mpx camera) and binning 4x 120fps (484*363) for 2.8 mpx camera while  30fps (512*512) for 4 mpx camera 

4- how the pixel size affect imaging one has 13*13 um while other one has 7*7um pixels. 

thanks.

The CCD camera of the given pixel captures of the image. I want calibrate the image and find the constant magnification factor for all the images.

Hello,

I am recording with a CCD camera epifluorescence of targeted cells on slices. Specific cells express a Gcamp, so we can follow the calcium variations through fluorescence variations.

For the analysis I extract intensity data of different ROIs with ImageJ and analyse the resulting curves in Igor. The problem comes with the fact that my fluorescent marker bleaches.

At first I would try to fit my data-baseline with exponential equations, or double exponential in the Neurmatic panel. But when the traces are too noisy, especially in the beggining or in the end, their is no way I could get a correct fit.

I have recently decided to change my approche and fit my baseline with an Akima's interpolation. But in calcium imaging, I did not see any papers using this fitting method.

Looking at my data, I really do feel like this method fits best my baseline (so does my boss and collegues). But I have no confirmation that this is the proper way to do it. I don't want to find out this is an aberant way of fitting the baseline.

Could anybody enlighten me?

I am trying to measure the electron beam damage induced from TEM on crystals using their diffraction spots and how they change over time. I know I can measure the difference in their intensity and their distance (r) but I don't know any other methods I could use. And how exactly do I finalize these results.

Also beam damage measurements from bright field images could be helpful for the future so I would appreciate any help!

Actually I have two interferogram ( containing Young's fringes). One interferogram is recorded without any perturbation in the path of light rays. While recording the second interferogram, I have kept a glass plate in the path of one light beam. Now I want to get information about refractive index of the glass plate by analysing the interferogram. Both Interferogram have been recorded usin a CCD camera. Can anyone help me?

I used beam profiler like the camera-based Cinogy beam profiler and the BP209 beam profiler from Thorlab to measure the beam profile of laser output from DFB laser’s output fiber directly. The beam profile is a perfect Gaussian shape. And then I used an industrial CCD camera to replace the original beam profiler. The beam profile is kind of different. It has some stripes which doesn’t changed much even I adjust the position and layout of the output fiber and cut the fiber over and over again. It looks like some kind of interference but I don’t know exactly how it occurs and why it only visible to the industrial CCD camera, not the professional beam profiler. Anyone can explain it? The attached files are the beam profilea captured by the CCD camera and the spectrum of the DFB laser.