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顕微鏡 Nikon C1siのPDF ユーザーマニュアルをオンラインで閲覧またはダウンロードできます。Nikon C1si 7 ページ。 Spectral laser scanning confocal microscope
ページを印刷する7. Turn on computer:
8. Launch software (EZ-C1 3.80).
a. Choose the colors you are going to use: 408/488/561, 488/561/638, or
457/514 (If you plan on doing spectral imaging it doesn't matter what you
choose in the software).
9. Choose standard or spectral detector.
a. Standard detector: Flip lever on side of the scanhead diagonal.
b. Spectral detector: Flip lever on side of scanhead vertical.
10. Choose excitation/emission wavelengths and put in scanhead dichroics and filter
cubes:
Beam splitter
(spectral detector)
Blue/Green/Red
Green/Red/Far Red
CFP/YFP
11. Align pinhole for your particular scanhead dichroic that you have selected:
a. Place a green fluorescent slide under your objective and focus on it using the
arc lamp and the eyepieces (you 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 pixels, green laser and green detector are on, and then go live.
d. Turn up the gain on the green detector until you get signal.
e. Use a 2.5mm hex key to align the pinhole using the two screws on the bottom
right of the scan head.
i.
This is done by 'walking' the pinhole: Move one screw until it gets as
bright as possible, then move the other screw to the brightest point, return
to the first one, and repeat until you get the brightest image possible. If
the image gets too bright so that all the pixels are saturated just turn
down the detector gain.
12. Take off the fluorescent slide and start imaging!
13. If using transmitted light, turn on halogen lamp (Lamp with dial on shelf) or turn off
for fluorescence.
Scanhead Dichroic
BS 20/80
408/488/561
488/561/638
457/514
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Left Filter Cube
n/a
595/50
525/50
685/70
595/50
550/50
485/30
Right Filter Cube
n/a
450/40
525/50