Leica TCS SP8 MP Скачать руководство пользователя страница 16 | Manualshive

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Objectives

Objectives may be selected by the pull-down menu. The

objectives will be raised and lowered as part of the

rotation. The multi-function plate (MFP) should be set to

substrate

for most applications. Use Automatic

determination of the beamsplitter.

Selecting the Fluorescence Detectors

Up to four detectors may be selected simultaneously. The

detector type (HyD or PMT) is indicated in the top left of

each detector window. The detector is selected by

clicking on the on/off button. The color of its display

output is selected from a pull-down menu next to the
type indicator. HyD detectors have a gain selected by

clicking on its slider button. PMT detectors have both

gain and offset functions, controlled by clickable sliders.

Leica recommends a starting offset of -0.3%.

Modifiying the Detector Bandwidth

The detector bandwidth may be modified in either

direction by simply clicking on and dragging its edges.

The minimum bandwith is 5 nm. Adjacent detector

bands may not overlap.

Selecting the Transmitted Light Detector

The TL detector is selected by clicking on its ON/OFF box. It is

a PMT and is responsive up to 900 nm. Like the other PMTs it

has gain and offset controls. Proper centering of the condenser

aperture is required for optimum effectiveness. The condenser

need not be focused but it should be close to the specimen.

Use the Dodt controller to modulate the contrast. The

optimum Dodt setting when viewed with the eyepieces is not

necessarily the same as the optimum setting when using

confocal.

Acquisition Start Buttons

At the base of the pane are four action buttons. At left, the

Autofocus

button should not be used. The

Live

button initiates

scanning at the indicated rate and format until it is pressed

again.

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Содержание TCS SP8 MP

Страница 1: ...ch as stage translation condenser height and objective rotation Thank you and we wish you every success in your imaging We are here to help The Confocal Microscope and Accessories The instrument is a...

Страница 2: ...confocal microscopy These detectors are electrically and liquid cooled for decreased background They should only be used in a darkened room and with the microscope covered by a dark room cloth availab...

Страница 3: ...wn focusing drive controlled from the microscope controller or the confocal controller or from the software The smaller Super Z Galvo stage rides on top of the large stage and includes the slide holde...

Страница 4: ...rcent excitation is required in most circumstances Optics The available objectives illustrated above from left to right are as follows Position Objective Magnification n a Working distance mm Medium 1...

Страница 5: ...via two screws as in the Zeiss system The condenser focus knob is to the left side The condenser cannot normally be brought into focus but as long as it is properly centered it will function satisfact...

Страница 6: ...cope controller are as follows Screen 1 Status and Illumination Use to adjust illumination intensity and the aperture and field stops in transmitted light TL These are in the microscope body not the o...

Страница 7: ...button to move the nosepiece to any position Note that this button will allow the nose piece to travel below the focus position Careless use of it may drive the objectives into the specimen The Home a...

Страница 8: ...roscope controller raise the nosepiece all the way to the top or to a level that allows you to change your sample without interference from the objectives Note the Set Clear Limits button bottom left...

Страница 9: ...til the specimen is in focus through the eyepieces with the 10X lens Raise it a small amount On the Set Clear Limits button push the Up Down button it will turn red The button will now be labelled Set...

Страница 10: ...e clips are positioned at the edges of the slide recess If the clips are hard to move use the hi tech tool provided Place the slide in the slide recess Usually placing the right side first is the most...

Страница 11: ...start LAS AF if the confocal power strip is not on There are two questions to be answered during startup Optional selection of multi photon laser Select MP_LASER_OFF Initialization of the large X Y st...

Страница 12: ...tor combination The knobs and buttons can be operated while acquiring in Live mode Modifications to them will be reflected in the Acquire pane From left to right they are Left button move the current...

Страница 13: ...s The lasers may also be set to Standby mode if not required for a period The Hardware dialog is useful to select the acquisition bit depth The default is 8 bit which is adequate for image acquisition...

Страница 14: ...s are available to all users from the pull down menu above Please don t modify them You can also create your own configurations and store them Acquisition Mode The acquisition mode defaults to xyz wit...

Страница 15: ...der The image area may also be translated in the directions indicated by the rosette arrows Large lateral translations at low zoom will move the scan range off axis and are therefore not recommended M...

Страница 16: ...set of 0 3 Modifiying the Detector Bandwidth The detector bandwidth may be modified in either direction by simply clicking on and dragging its edges The minimum bandwith is 5 nm Adjacent detector band...

Страница 17: ...ed to display the results may be changed using the button at the top left of the display window The first click selects a range finding LUT for the active window that can be used to set gains and offs...

Страница 18: ...be modified by clicking on Nr of Steps or z step size The diagram indicates the size and spacing of the stack and the current stage position It may be re scaled for better viewing using the slider pr...

Страница 19: ...ditions Acquisition conditions within the ROI will be as specified in the Detectors panel Draw the ROI using the tools above the image window In this example a square ROI has been selected To set the...

Страница 20: ...hem Up to six sequential acquisition events are possible Events may be any of the available modes e g xy xyz xyt xy Events may be acquired line by line indicated by arrow This is the preferred method...

Страница 21: ...21 Alternate Acquisition Modes Line Scan...

Страница 22: ...nge Properties window The maximum spectral range is 380 nm to 785 nm The shortest wavelength laser is 405 nm so the shortest practical wavelength is 420 nm unless multi photon excitation is used The m...

Страница 23: ...23 Alternate Acquisition Modes Time Series Select xyt or similar acquisition mode...

Страница 24: ...rent position for storage Multiple positions can be stored The current position number is reflected below the map The second button deletes the current stored position as reflected by the position num...

Страница 25: ...e scanned In this example a 2 x 1 tile map is to be acquired Stage 3 To expand the tiled area move the stage to a new position at an orthogonal extreme as before Click on the Select Position button to...

Страница 26: ...window will increment You may delete the current position using Delete Current Position or delete all currently defined positions using the trash can You may also re define the current position A fami...

Страница 27: ...xperiments Double click on an experiment to inspect the result or analyze it Right click on an experiment or sub experiment to change its name Use the Save button below in the Experiments window to sa...

Страница 28: ...the experiment if it is not already loaded Click on the sub experiment Select the Apply button at the base of the Experiments window to apply the parameters If you wish to check the parameters first S...

Страница 29: ...ton laser is on Multiphoton gain and offset All detector settings including acquisition type for HyDs Standard vs Photon Counting Pinhole diameter These parameters are NOT applied Bit depth important...

Страница 30: ...30 Process Pane...

Страница 31: ...31 Quantify Plane...

Страница 32: ...ary to power down the computer or the microscope when switching from MP_LASER_OFF to MP_LASER_ON operation However certain steps must be followed for the correct sequence Switch on the Electro Optical...

Страница 33: ...ing the MP Laser Unlike the semiconductor lasers the MP laser must be turned on in the Configuration pane Setting up the MP Laser In the Acquire pane first click MP on Then click on the button to open...

Страница 34: ...SER_ON mode will enable the NDD Super HyD detectors These are controlled using a single gain slider The NDD detectors are very sensitive Use low gain and low laser power initially Operate only in a da...

Страница 35: ...aser intensity at the specimen The number at the base of the left slider is the excitation wavelength in nm Best practice is initially to set the coarse slider to zero and the fine slider to a low val...

Страница 36: ...f the computer Switch off the confocal at the power strip Don t forget to restore the Home and Focus levels if you have changed them Replace the condenser and any non standard objectives in the drawer...

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