Multiphoton Lasers and Fluorescence Filters
Lasers
This microscope is equipped with a 405nm diode, a 561nm DPSS, a 633 HeNe, and an Argon laser with lines at 458nm, 488nm and 514nm. It also has a Ti:Sapphire laser tuneable between the wavelengths of 720-1064nm, and the OPO is tuneable between 1050-1300nm.
Dichroic Mirrors for Invisible Light
Position | Dichroic | Example Fluorophores | Notes |
|---|---|---|---|
1 | Plate |
| Use this when not using any Invisible Light lasers |
2 | Mirror |
|
|
3 | MBS 405 | DAPI, Hoechst |
|
4 | MBS 445 |
| Not used |
5 | MBS 690 | Various | Use this for multiphoton laser ablation at <760 nm as well as imaging |
6 | MBS 760 | Various | Use this for multiphoton imaging >760 nm |
7 | MBS 405(UV)/760 | Various | Use this for combining excitation with the 405 nm laser with the multiphoton (>760 nm) |
8 | MBS 445/760 |
| Not used |
9 | MBS T80/R20 |
| Use for reflection imaging |
10 | None |
|
|
Dichroic Mirrors for Visible Light
Position | Dichroic | Example Fluorophores | Notes |
|---|---|---|---|
1 | MBS 458 | CFP |
|
2 | MBS 458/514 | CFP/YFP |
|
3 | MBS 458/561 | CFP/RFP |
|
4 | None |
|
|
5 | MBS 488 | GFP, Alexa 488 |
|
6 | MBS 488/561 | GFP/RFP, Alexa Fluor 488/Alexa Fluor 568, FITC/TRITC |
|
7 | None |
|
|
8 | MBS 488/561/633 | GRP/RFP/Alexa Fluor 633 |
|
9 | MBS T80/R20 |
| Use for reflection imaging |
10 | Plate |
|
|
Filter sets in the microscope stand reflector revolver
Position | Filter Set | Notes |
|---|---|---|
1 | FSet49 wf | Zeiss Filter Set 49. Bandpass filter set for viewing DAPI and similar blue fluorophores down the eyepieces. |
2 | FSet38 wf | Zeiss Filter Set 38. Bandpass filter set for viewing Alexa 488, eGFP and similar green fluorophores down the eyepieces. |
3 | FSet43 wf | Zeiss Filter Set 43. Bandpass filter set for viewing TRITC, DsRed and similar orange fluorophores down the eyepieces. |
4 | BS-MP 355/690+(R) | Only for use with multiphoton. Reflects laser light >690 nm, and transmits light <690 nm to the non-descanned detectors. |
5 | BS-MP -760 | Only for use with multiphoton. Reflects laser light >760 nm, and transmits light <760 nm to the non-descanned detectors. |
6 | Rear | For confocal imaging. Sends all light to the scan head. |