Leginon

From Gaya Yadav

I Aligned different presets using "en" as reference they looks aligned well 
but when i select " exposure targets in the hole, the image captured is on the carbon
so could you please suggest which parameter i need to calibrate to make it happen in hole?

The alignment presets make sure that if you move stage at one preset to put a feature to the center of the camera, the image taken in other preset will also have the feature at the center.

The calibration that relates the scale and direction of a microscope operation like "stage position", "image shift", "beam shift" to pixel vectors on camera determines the other half of targeting, i.e. how much and in what direct it should tell microscope to move in order to put the feature you want (your target) to the center of the camera.

Therefore, you should check the matrix calibration of the move method you defined in "Exposure" node. In your setup, it is likely "image shift". To map what need to be move at the "en" preset, both image shift matrix in "hl" preset where the target is selected from and image shift matrix in "en" preset magnification are used.

Next, to map relative orientation of the image shift coil in the two preset, we need a reference space which will be the stage position calibration. I am planing to make it easier in the future, but for now, what you should do is to get a reasonable stage calibration of the high mag (i.e., en). This is hard to do due to the small area it covers so the trick with most modern microscope is to scale a lower mag one using the instruction in Scaling Stage Position Matrix Calibration