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14. As needed, engage the Z-stepper-motor (used to do Z-stacks) by pulling up gently 

(on back right of microscope). 

 
Shutdown:  

1.  If no one is signed up 2 hours after your session is over:  

a.  Raise objectives fully up and move up the focus. 
b.  Clean off oil objectives with lens paper and replace lens in its holder. 
c.  Reverse of Turn on, items 6

1. 

2.  If someone is signed up after you:  

a.  Turn off EZ-C1 software.  
b.  Leave everything else on.  

 

Overview 

 

The C1si is a confocal laser-scanning microscope with four lasers for multi-color 
imaging.  It has 4 lasers with these laser lines: 408nm, 457nm, 488nm, 514nm, 561nm, 
638nm.  It has 2 types of detectors: one with 3 standard band pass detectors and one 
with a spectral detector that can image 32 different channels at the same time (they can 
be set to be 2.5nm, 5nm, and 10nm wide). It is on an upright Nikon FN1 microscope with 
a large electrophysiology stage. The max penetration depth into tissue is approximately 
100–200 µm depending on the opacity of the sample. 
 
To move the objective turret between the two lens positions, center the lever so that both 
lenses are in the up position and then press the button on the upper right part of the 
carriage and move forward or back. Make sure that it is in the very center so that when 
you change positions it does not fall down and get damaged.   

Note: the viewed area may change because of slight differences in lens position 
between the two holders. 

 

Summary of Contents for C1si

Page 1: ...Microscope User Guide Contents C1Si Turn On ShutDown Procedures 2 Overview 4 Setup for epi illumination to view through the eyepieces 5 Setup for confocal imaging 5 Notes 6 Troubleshooting 7 Owners Consortium Department of Pathology SABRE Proctor Foundation ...

Page 2: ...the up position Use both hands and gently thread it until it is finger tight Do not over tighten it or it will get stuck Available Objectives Dipping objectives are designed to form an image without a cover slip whereas immersion lenses require a cover slip For ideal imaging always use 1 5 cover slips You can then switch between the two different objectives that you have loaded by turning the leve...

Page 3: ...ou need to push the eyepiece slider in open the shutter and choose the GFP filter cube it gets very bright so you can put in the neutral density filters on the back right of the scope so you don t blind yourself b Engage the confocal pull eyepiece slider out close shutter and move filter cube to open position 6 c Turn the software onto live and set it up so the color changes with intensity 256x256...

Page 4: ...It has 2 types of detectors one with 3 standard band pass detectors and one with a spectral detector that can image 32 different channels at the same time they can be set to be 2 5nm 5nm and 10nm wide It is on an upright Nikon FN1 microscope with a large electrophysiology stage The max penetration depth into tissue is approximately 100 200 µm depending on the opacity of the sample To move the obje...

Page 5: ...ckground is black a If you are using frame lambda set up each pass individually b Set your pinhole generally it is best to leave it at a small 30 micron pinhole unless there is a compelling reason to change it 6 If using the standard detector set the detector gains at 6 5 your usable range is generally between 5 and 8 7 Go live and slowly turn up the laser power to your sample and then increase de...

Page 6: ...olution limit You know you have reached this because the note not optimum pixel size disappears 11 XY basic a Pixel dwell time this is how long the laser stays on one pixel I usually leave this at the minimum time b Field zoom the zooming that you have done in step 10 c Pixels I recommend using either 512x512 or 1024x1024 but you can do arbitrary if you want 12 Time a Use this to set up your time ...

Page 7: ...s set to 6 The eyepiece scanhead changer is pulled out Both the detector and laser are turned on so they are colored The detectors that you want are moved to 6 5 or higher The sliders for the Lasers that you want are on Your sample is getting photo bleached too quickly Try using less laser power and more detector gain Try focusing using single frames instead of live Your image is too noisy Try inc...

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