prairie
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prairie [2025/01/10 17:49] bioimaging |
prairie [2025/01/11 17:06] (current) bioimaging |
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| ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ Fluorochromes ^ | ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ Fluorochromes ^ | ||
| - | | 2p (pos1) | - | NIR | - | mirror | all | | + | | 2p (pos1) | - | mirror | - | - | all | |
| | Green (pos3) | - | blue | - | ~525 nm | GFP, FITC, Alexa488 | | | Green (pos3) | - | blue | - | ~525 nm | GFP, FITC, Alexa488 | | ||
| | Orange (pos4) | - | green | - | ~580 nm | TRITC, Alexa568 | | | Orange (pos4) | - | green | - | ~580 nm | TRITC, Alexa568 | | ||
| Line 61: | Line 61: | ||
| \\ | \\ | ||
| ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ||
| - | | Ch1 (PMT) | Far Red | 640-680 nm | Alexa647, Cy5 | | + | | CH1 (PMT) | Red | 640/680 nm | Alexa647, Cy5 | |
| - | | Ch2 (GaAsP) | Orange | 570-620 nm | mCherry, Alexa568, Cy3 | | + | | Ch2 (GaAsP) | Green | 570-620 nm | mCherry, Alexa568, Cy3 | |
| - | | Ch3 (GaAsP) | Green | 500-550 nm | GFP, Alexa488, FITC | | + | | Ch3 (GaAsP) | Far Red | 500-550 nm | GFP, Alexa488, FITC | |
| - | | Ch4 (PMT) | Blue | 435-485 nm | DAPI, CFP or BS 2nd Harmonic | | + | | Ch4 (PMT)* | Blue | 435-485 nm | DAPI*, CFP or SHG | |
| - | | Ch4 (DODT) | BF | - | Brightfield | | + | |
| - | *1 note: Ch4 can be used for either DAPI, BF or 2nd Harmonic. | + | |
| + | *Ch4 can be used either in fluorescence mode, or alternatively as a BF ("DODT") channel or for SHG - ask staff | ||
| - | + | \\ | |
| - | ===== System Turn On Procedures ===== | + | **Upon request:** |
| + | \\ (requires changing filters and the dichroic beamsplitter) | ||
| UNDER CONSTRUCTION!!! | UNDER CONSTRUCTION!!! | ||
| - | * 1. Check that the cooling and power supply for the laser are ON {{::img.png?100|}} | + | ===== System Turn On Procedures ===== |
| - | * 2. Turn on the fluorescence lamp {{1_img.png?100|}} | + | |
| - | * 3. ... | + | * Check the cooler. The temperature should be 20 Cº. {{::img.png?100|}} |
| + | * Turn on the fluorescence lamp {{1_img.png?100|}} | ||
| + | * Switch the laser from "stand by" to "on" by turning the key clockwise (unit under the table) | ||
| + | * Make sure the workstation is ON | ||
| + | * 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.1736527755.txt.gz · Last modified: 2025/01/10 17:49 by bioimaging
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