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Iron Man

Zeiss LSM 510 Laser Scanning Confocal Microscope

User Guide

v. 1.4 (11/2016)

Objectives

5x/0.15NA EC Plan Neofluar
10x/0.3NA EC Plan Neofluar
20x/0.8NA Plan Apo Chromat

40x/1.3NA

oil

DIC EC Plan Neofluar

63x/1.4NA

oil

DIC Plan Apo Chromat

100x/1.4NA

oil

DIC Plan Apo Chromat

Standard coverslip thickness 0.17
(#1.5 coverslip)

Mercury Arc lamp (ocular)

Dapi, Fitc, Rhod, DIC II, DIC III, Brightfield

Laser lines

Argon/2 –

458

477

488

514

HeNe –

543

HeNe –

633

Diode –

405/30

Содержание LSM 510 Inverted

Страница 1: ...EC Plan Neofluar 20x 0 8NA Plan Apo Chromat 40x 1 3NA oil DIC EC Plan Neofluar 63x 1 4NA oil DIC Plan Apo Chromat 100x 1 4NA oil DIC Plan Apo Chromat Standard coverslip thickness 0 17 1 5 coverslip Me...

Страница 2: ...Tab 6 At the top In the Setup Manager window In the Imaging Setup window In the Channels window In the Online Acquisition window Adjusting PMTs Snap Z series The Information tab next to the image 11 W...

Страница 3: ...ment 8 Note the details in Information tab next to your image these are useful to write in your imaging notes 9 Save and copy your data The LMCF is not responsible for long term data storage See Appen...

Страница 4: ...including lasers 2 Mercury Arc lamp located underneath floating air table 3 PC 4 The microscope green switch on right side of microscope base If you only need to access the PC only turn on 1a and 3 L...

Страница 5: ...u are not looking through the eyepieces To look at your slide 1 Load the slide coverslip down onto the slide holders on the stage these can be adjusted manually Please be sure that your slide is clean...

Страница 6: ...when it is done Choose On Open the Laser Properties window by clicking on the arrow and adjust laser output to 40 Do not go over 40 output or over 6 1 A Do not keep changing this Output set it for yo...

Страница 7: ...k will be faster but may lead to bleedthrough of your signal across channels An example of both a single track multi channel acquisition and multi track acquisition are shown To add or remove tracks c...

Страница 8: ...ges and can allow you to reduce your scan speed Bit Depth the number of grey levels the image will contain Higher bit depth will result in greater signal dynamic range greater discrimination between p...

Страница 9: ...ing values for the Argon and Diode lasers 3 5 is usually sufficient and for the HeNe lasers 25 35 is usually a good starting point Set the pinhole size to 1AU Unless you have a deliberate reason to ch...

Страница 10: ...ion until desired distance is reached click set last o Repeat in the other direction click set first o Either choose number of slices or step size The software will suggest the optimal step size This...

Страница 11: ...objectives you may have used with lens paper If you are at all unsure of this process ask for help 3 Leave the microscope on a low power objective for next user If someone is signed up within the next...

Страница 12: ...e and password 5 Click Ok and the drive should now appear Copy over your files If you have a lot of files be sure to allow time for the transfer Under ideal conditions you will be able to copy close t...

Страница 13: ...Iron Man User Guide v 1 4 13 APPENDIX B Parts of the microscope not all these components are present on our microscope...

Страница 14: ...o bleedthrough of signal You can do a combination of multi track and multi channel for example image DAPI and red together in a mult channel track and image green separately in its own track Load a pr...

Страница 15: ...Resources tab at the bottom Life Technologies SpectraViewer is a straighforward and reasonably complete one Excitation spectra are shown as dotted lines and emission spectra are solid Next figure out...

Страница 16: ...transmit the emission from the sample through 405 488 405 514 594 405 488 543 633 458 458 514 488 543 488 594 NFT 80 20 neutral density filter passes 80 and blocks 20 2 Sample 3 Dichroic split emissio...

Страница 17: ...l reflect all light below 543nm to the right and allow to pass through all wavelengths greater than 543 The following plot illustrates the emission spectra of our three fluorophores with a green line...

Страница 18: ...nels It is important to note the overlap in emission spectra In this example it is possible that some of the DAPI emission will show up in the FITC channel and some FITC emission will show up in the R...

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