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h2. Leica TCS SPE confocal microscope (SPE3)


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*Location:* *MRC Building, Room 2.16*

{float:right}!SPE3.jpg|border=1!{float}

The SPE3 is a second generation Leica TCS SPE confocal microscope mounted on a semi-automated, inverted Leica DMI4000 stand. The stand is capable of fluorescence, bright-field and DIC techniques and is also fitted with a motorised stage for multi-point (Mark & Find) and stitching (Tile Scan) acquisitions. The confocal system has a single photomultiplier tube (PMT) detector so all multichannel acquisition must be done sequentially. The ACS objective lenses have been designed to correct for  colour registration errors between the violet and visible lasers. They do not have a  flat field so Leica recommend that the zoom is set to ≥1.6. There is also the option on this machine to carry out small screens thanks to a matrix screener software.

| *Contents* |
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h3. Access & Usage
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* [Training|LMCBLMic:LMCB Microscope Training]
* [Confocal Booking System (Faces Scheduling System)|http://faces.ccrc.uga.edu/]
* [Microscope Booking Rules|LMCBLMic:Microscope Booking]

h3. Technical Specifications
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* [Microscope Stand Information|SPE3 Microscope Information]

h2. Lasers and Detectors

* {color:#800080}405 nm{color} {color:#000000}(violet, for blue dyes like DAPI){color}
* {color:#00ccff}488 nm{color} {color:#000000}(blue, for green dyes like GFP){color}
* {color:#99cc00}561 nm{color} {color:#000000}(lime green, for red dyes like mCherry){color}
* {color:#ff0000}635 nm{color} {color:#000000}(red, for far red dyes like Cy5){color}

It has a quad dichroic mirror meaning a rapid switch between channels.

There is only one photomultiplier tube (PMT) so all multi-channel acquisition is done sequentially

A galvanometer driven stage insert is used to change the focus when optically sectioning samples. The speed of this device means that XZY scans are possible in addition to the conventional XYZ scan.

The ACS objective lenses, which have been designed to correct for colour registration errors caused by chromatic aberration, do not have a flat field, so best results are achieved at zooms ≥1.6.


There is also the option on this machine to carry out small screens thanks to a matrix screener software

The following PDF lists the recommended start-up and shut-down order for the machine: [^SPE3_StartUp_Shut Down.pdf]
















h4. Changing the Galvo Z stage inserts

There are three inserts for the Galvo Z stage, although in practice you'll probably only need to use two at most. The main ones are the universal adjustable insert that will take slides, dishes, Lab-tek chambers etc and the multi-well plate insert. The universal insert should be fitted by default since it suits the most users. _If you use the insert for the multi-well plate then you_ {color:#ff0000}{_}MUST{_}{color} _replace the universal adjustable stage insert once you have finished_. The screws for the universal insert are very small so you MUST be careful not to drop any into the microscope stand; particularly under the objective lens nose-piece. If a screw rolls under the nose-piece then contact a member of light microscope staff immediately.

h3. Links:

* [Microscope Information|SPE3 Microscope Information]
* [Transmitted Light DIC on SPE3|^TransmittedLightImagesontheSPE.pdf]
* [Confocal Booking System|http://faces.ccrc.uga.edu]
* [Confocal Resolution Calculator|http://lightmicroscopy.lmcb.ucl.ac.uk/Information/LSM_Res.html]