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Zeiss LSM 880 with Airyscan

Confocal Light Microscopy

Basic User Guide

Содержание LSM 880

Страница 1: ...Zeiss LSM 880 with Airyscan Confocal Light Microscopy Basic User Guide...

Страница 2: ...tors 7 Transmitted Light Techniques 7 Incident Light 7 Features and Specifications 7 Objective 8 Filter Sets for Incident Light 9 Additional Equipment 10 Computer 10 Monitor 10 Potential Techniques 10...

Страница 3: ...3 Smart Setup 17 Lasers 18 Imaging Setup 18 Acquisition Mode 21 Channels 22 Definite Focus 2 23 Multidimensional Acquisition 23 Z Stack 23 Time Series 24 Tile Scan 24 Positions 25 Regions 25...

Страница 4: ...350 nm Z even in thick samples The introduction of a Fast module for the Airyscan offers a satisfactory compromise between perfect optical sections and live cell imaging experiments The Fast Airyscan...

Страница 5: ...enide phosphide GaAsP detector a 63x Plan Apochromat 1 4NA DIC oil immersion objective Zeiss and the 405 nm 488 nm and 561 nm laser lines The Zeiss Zen 2 3 black edition software was used to control t...

Страница 6: ...pment cannot be used without official training Before your first use contact Mariana Ferreira to schedule training Save all data into your own storage device as we routinely clean the computer s data...

Страница 7: ...ast DIC Phase Contrast Bright Field Incident Light Fluorescence HXP UV lamp Features and Specifications Axio Observer 7 SP Inverted Microscope Stand Motorized Stage 130 x 100 STEP Stage Controller XY...

Страница 8: ...nterference Contrast DIC II Differential Interference Contrast DIC III Lateral Resolution XY 525 nm m 2 0 7 0 2625 0 15 Nyquist Sampling XY 525 nm m 3 0 28 0 105 0 06 Zoom to Nyquist Sampling XY 525 n...

Страница 9: ...he Locate Tab Emission Filters for Airyscan and Fast Airyscan Position Wavelengths Emission Colours Optimal Fluorophores 2 BP 420 480 BP 495 550 Blue Green to Yellow DAPI GFP FITC YFP 3 BP 420 480 BP...

Страница 10: ...12 free slots 4x 2TB 4 TB RAID 10 NVIDIA Quadro K2200 4GB OS Windows 7 Ultimate Embedded x64 Available Software Zeiss Zen 2 3 Monitor Brand HP Model DreamColor Z32x Diagonal 32 Native Resolution 4K UH...

Страница 11: ...plore your fluorophores Learn their excitation and emission wavelengths and what are their optimal laser lines and filters This is necessary to use the confocal to its full potential Sample Preparatio...

Страница 12: ...see the Windows start up screen 5 Turn on the Components Switch 6 Wait for everything to finish initializing 7 Open Zen Black on the computer 8 Choose Start System and wait for the initialization to...

Страница 13: ...laser in Standby mode and turn all other lasers off b Otherwise turn all lasers Off 5 If turned Off it will take the Argon Laser at least 5 minutes to cool fans turn off 6 Shutdown the computer and th...

Страница 14: ...menu bar above and a system information bar below The Image and Document Manager Section shows all the currently open files and allows you to save and close the currently selected image The Image Disp...

Страница 15: ...transmitted light and off The transmitted light setup is used for every brightfield configuration with the type brightfield phase contrast or DIC being dependant on the condenser setting and prisms o...

Страница 16: ...live feed Snap Acquire current plane with your settings Additional available acquisition options check to show options tab To acquire images with these extra options you must use Start Experiment ins...

Страница 17: ...ee possible regular confocal configurations are generated with optimization for speed Fastest for Best Signal and a compromise between both Smartest Each configuration will display the speed percentag...

Страница 18: ...d The Argon laser takes a few minutes to warm up Imaging Setup At the top of this tab you have Tracks There can be up to four tracks simultaneously each with their own imaging mode and lightpath setup...

Страница 19: ...or strong fluorophores with Ch3 detector having an extra sensitivity to the red part of the spectrum 5 5 You can also identify the fluorophore from the dropdown list like in Smart Setup and assign its...

Страница 20: ...20 2 3 4 7 8 5 6 9 1...

Страница 21: ...Airyscan requires Optimal frame size Higher speeds give noisier images The recommended speed is 5 or 6 Check the Direction is bi directional as it is faster Use either 8 Bit or 16 Bit for Bit Depth p...

Страница 22: ...t to the histogram in the display area to see if you need to adjust the laser power If you are using the full dynamic range a lot of RED pixels means you are saturating the detector all max intensity...

Страница 23: ...g media such as Vectachield will be suboptimal unless diluted with an aqueous buffer To use this first select Definite Focus in the Focus Devices and Strategy tab then in the Definite Focus tab click...

Страница 24: ...te Focus in longer time lapses to ensure the sample stays focused Number of cycles is the number of pictures to take if zero is selected the software will take pictures until manually stopped Interval...

Страница 25: ...of the current frame will be saved Regions The pixel by pixel image acquisition of the confocal allows you to acquire only Regions Of interest ROIs instead of the whole frame You can make ROIs using...