prairie
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prairie [2025/01/09 18:21] bioimaging |
prairie [2025/01/11 17:06] (current) bioimaging |
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| <a href="/facility/bioimaging/lib/exe/fetch.php?media=prairie.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=prairie.jpg" width=300></a> | <a href="/facility/bioimaging/lib/exe/fetch.php?media=prairie.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=prairie.jpg" width=300></a> | ||
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| - | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_map_new.png">[Oeiras] 0B06 </a><br><b>Manufacturer</b>: <a href="https://-/" target="_blank">Prairie (now Bruker!)</a><br> | + | <b>Location</b>: [Oeiras] 0B06 |
| - | <b>Model</b>: <a href="https://" target="_blank">Prairie Ultima </a><br> | + | <br><b>Manufacturer</b>: Prairie (now Bruker!)<br> |
| + | <b>Model</b>: Prairie Ultima <br> | ||
| <b>Nickname</b>: "Prairie"<br> | <b>Nickname</b>: "Prairie"<br> | ||
| <b>Software</b>: PrairieView 4.5 <br> | <b>Software</b>: PrairieView 4.5 <br> | ||
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| <b>SN</b>: N/A <br> | <b>SN</b>: N/A <br> | ||
|   <br> |   <br> | ||
| - | Data will be deleted after: <b>3 months</b> | + | Data older than 3 months is automatically deleted!</b> |
|   <br> |   <br> | ||
|   <br> |   <br> | ||
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| ==== Objectives ==== | ==== Objectives ==== | ||
| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
| - | | 20x | Olympus (Evident now!) XLUM FL Plan | Water (dipping) | 1.00 | 2.00 | [[https://www.thorlabs.com/thorproduct.cfm?partnumber=N20X-PFH]] | | + | | 20x | Olympus (now Evident!) XLUM FL Plan | Water (dipping) | 1.00 | 2.00 | [[https://www.thorlabs.com/thorproduct.cfm?partnumber=N20X-PFH]] | |
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| ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ Fluorochromes ^ | ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ Fluorochromes ^ | ||
| - | | Green | - | 525/50 nm | blue | > 515 nm | GFP, FITC, Alexa488 | | + | | 2p (pos1) | - | mirror | - | - | all | |
| - | | Orange | - | 5xx/xx nm | green | > xxx nm | TRITC, Alexa568 | | + | | Green (pos3) | - | blue | - | ~525 nm | GFP, FITC, Alexa488 | |
| - | | Blue | - | 460x/xx nm | violet | > xxx nm | DAPI | | + | | Orange (pos4) | - | green | - | ~580 nm | TRITC, Alexa568 | |
| + | | Blue (pos5) | - | violet | - | ~460 nm | DAPI | | ||
| ==== Emission Filters (multi-photon) ==== | ==== Emission Filters (multi-photon) ==== | ||
| {{::xxfiltersetDiagram.png |}} | {{::xxfiltersetDiagram.png |}} | ||
| - | \\ | ||
| - | \\ | ||
| \\ | \\ | ||
| ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ||
| - | | CH1 (PMT) | Red | 660/40 nm | Alexa647, Cy5 | | + | | CH1 (PMT) | Red | 640/680 nm | Alexa647, Cy5 | |
| - | | Ch2 (GaAsP) | Green | 595/50 nm | mCherry, Alexa568, Cy3 | | + | | Ch2 (GaAsP) | Green | 570-620 nm | mCherry, Alexa568, Cy3 | |
| - | | Ch3 (GaAsP) | Far Red | 525/50 nm | GFP, Alexa488, FITC | | + | | Ch3 (GaAsP) | Far Red | 500-550 nm | GFP, Alexa488, FITC | |
| - | | Ch4 (PMT1) | Blue | 460/50 nm | DAPI, CFP or BS 2nd Harmonic | | + | | Ch4 (PMT)* | Blue | 435-485 nm | DAPI*, CFP or SHG | |
| - | \\ | + | |
| - | \\ | + | *Ch4 can be used either in fluorescence mode, or alternatively as a BF ("DODT") channel or for SHG - ask staff |
| \\ | \\ | ||
| **Upon request:** | **Upon request:** | ||
| \\ (requires changing filters and the dichroic beamsplitter) | \\ (requires changing filters and the dichroic beamsplitter) | ||
| - | |||
| - | {{::alternative_filtersetDiagram.png |}} | ||
| - | \\ | ||
| - | \\ | ||
| - | \\ | ||
| - | ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ||
| - | | Ch1 | - | - nm | Cy5 | | ||
| - | | Ch2 | - | - nm | Alexa568 | | ||
| - | | Ch3 | - | - nm | Alexa488 | | ||
| - | | Ch4 | - | - nm | DAPI | | ||
| - | | DODT | - | - nm | BF | | ||
| - | \\ | ||
| - | \\ | ||
| - | \\ | ||
| UNDER CONSTRUCTION!!! | UNDER CONSTRUCTION!!! | ||
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| ===== System Turn On Procedures ===== | ===== System Turn On Procedures ===== | ||
| - | * 1. Check that the cooling and power supply for the laser are ON | + | * Check the cooler. The temperature should be 20 Cº. {{::img.png?100|}} |
| - | {{::img.png?100|}} | + | * Turn on the fluorescence lamp {{1_img.png?100|}} |
| - | * 2. Turn on the fluorescence lamp | + | * Switch the laser from "stand by" to "on" by turning the key clockwise (unit under the table) |
| - | {{1_img.png?100|}} | + | * Make sure the workstation is ON |
| - | * 3. ... | + | * Turn on the pre-amps for the detectors you will use (Ch2 and Ch3 pre-amp unit in shelf above monitor, and Ch1 and Ch4 below the keyboard. Note that any switching of the Ch1&2 pre-amps should be preceded by "override" for protetion). |
| + | * Ask staff to change the Ch4 for DODT (BF) detector or fluorescence/SHG | ||
| ===== System turn Off procedures ===== | ===== System turn Off procedures ===== | ||
| + | |||
| + | * Transfer your data to the institutional server (data is deleted every 3 months) | ||
| + | * Clean up immersion objective(s) | ||
| + | * Remove all your samples and accessories and leave the working area ready for the next user | ||
| If there is another user for this microscope in the __next hour__: | If there is another user for this microscope in the __next hour__: | ||
| - | * Leave the fluorescent lamp and lasers on | + | * Leave the fluorescent lamp and LASER "ON" |
| - | * Clean up immersion objectives | + | * Inform staff you are leaving the system "ON" |
| + | |||
| Else: | Else: | ||
| - | * Clean up objective(s) | + | * Set the LASER to "Standby" (LASER key to the left) |
| - | * Set the laser to ''standby'' | + | * Turn "OFF" all pre-amps (make sure to protect 1st for Ch1&2! |
| - | * ... | + | * Turn "OFF" the fluorescence lamp |
| + | * Turn "OFF" monitor, lights, and close the microscope curtains | ||
| + | * Make sure you turn "OFF" any life support unit used (heating, CO2, O2...) | ||
| ---- | ---- | ||
| [[resources|Back to the Equipment page]] | [[resources|Back to the Equipment page]] | ||
prairie.1736443288.txt.gz · Last modified: 2025/01/09 18:21 by bioimaging
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