Zeiss LSM 510 META Manual de usuario descargar pdf (Pagina 6)

Zeiss LSM 510 Meta Confocal Microscope Instructions

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drop, waiting for you to add a drop of immersion fluid to the coverslip. After adding immersion

fluid, tap Done

(if you change your mind, Back will return you to the last

objective lens used).

 Only use Zeiss Immersol 518F with 40X and 63X oil-immersion objective lenses and either

distilled water or Zeiss Immersol W (for long-term work) with 40X water-immersion objective

lens.

e. To examine your sample by reflected light (RL) (i.e. exciting with

Hg lamp and observing fluorescence produced), turn TL off using TL

knob or TL button, then tap Reflector tab.

We have 3 filter sets for observing fluorescence through eyepieces,

indicated by buttons under Reflector tab labeled:

DAPI (blue fluorescence)

FITC (green fluorescence)

Rhodam[ine] (red fluorescence)

Tap on filter set button most appropriate to the fluorescence you wish to observe, then tap Open

under RL-Shutter to allow light from Hg lamp to reach slide. Tap Close once you have observed

fluorescence in area of interest and centered it in field of view.

 Open RL-shutter for as short a period as possible to reduce photobleaching of your fluor(s).

 Buttons under Reflector tab labeled Pos. 1, Pos. 6, and DIC TL are only used with TL (halogen

lamp); DAPI, FITC, and Rhodam[ine] are only used with RL (Hg

lamp). Pos. 1 and Pos. 6 are empty positions (contain no optical

elements), either of which should be selected for regular TL viewing.

DIC TL adds differential interference contrast (DIC) optics to the TL

lightpath to add contrast to otherwise virtually invisible samples.

However, the 5X and 10X objective lenses lack the additional DIC

optical elements needed for true DIC.

16. Collecting laser-excited confocal fluorescence images on monitors.

a. Click Find button in Scan Control window.

An Image display window will open.

b. If you are imaging two or

more fluors (3 in this

example), default is for all

channels to be displayed in a

single, overlapping image.

Click Split xy button to see

each channel in a separate

panel.

Fluorescence is first displayed

just for whichever channel

happened to be selected when

the

Find button was clicked. Click a different channel button,

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Zeiss LSM 510 META Manual de usuario Pagina 6

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