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Multiphoton Lasers and Fluorescence Filters

Multiphoton Lasers and Fluorescence Filters

Owned by Ki Hng (Deactivated)
Last updated: Jul 09, 2024 by Andrew Vaughan

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

PositionDichroicExample FluorophoresNotes
1Plate
Use this when not using any Invisible Light lasers
2Mirror

3MBS 405DAPI, Hoechst
4MBS 445
Not used
5MBS 690VariousUse this for multiphoton laser ablation at <760 nm as well as imaging
6MBS 760VariousUse this for multiphoton imaging >760 nm
7MBS 405(UV)/760VariousUse this for combining excitation with the 405 nm laser with the multiphoton (>760 nm)
8MBS 445/760
Not used
9MBS T80/R20
Use for reflection imaging
10None

 Dichroic Mirrors for Visible Light

PositionDichroicExample Fluorophores

Notes

1MBS 458CFP
2MBS 458/514CFP/YFP
3MBS 458/561CFP/RFP
4None

5MBS 488GFP, Alexa 488
6MBS 488/561GFP/RFP, Alexa Fluor 488/Alexa Fluor 568, FITC/TRITC
7None

8MBS 488/561/633GRP/RFP/Alexa Fluor 633
9MBS T80/R20
Use for reflection imaging
10Plate

Filter sets in the microscope stand reflector revolver

PositionFilter SetNotes
1FSet49 wfZeiss Filter Set 49. Bandpass filter set for viewing DAPI and similar blue fluorophores down the eyepieces.
2FSet38 wfZeiss Filter Set 38. Bandpass filter set for viewing Alexa 488, eGFP and similar green fluorophores down the eyepieces.
3FSet43 wfZeiss Filter Set 43. Bandpass filter set for viewing TRITC, DsRed and similar orange fluorophores down the eyepieces.
4BS-MP 355/690+(R)Only for use with multiphoton. Reflects laser light >690 nm, and transmits light <690 nm to the non-descanned detectors.
5BS-MP -760Only for use with multiphoton. Reflects laser light >760 nm, and transmits light <760 nm to the non-descanned detectors.
6RearFor confocal imaging. Sends all light to the scan head.


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