# \#tirf

**URL:** https://forum.microlist.org/tag/tirf/4.md

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## [Consult the excitation optical path of TIRF](https://forum.microlist.org/t/consult-the-excitation-optical-path-of-tirf/3054)

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**Author:** [@SLEEzZ](https://forum.microlist.org/u/SLEEzZ)\
**Replies:** 2\
**Last updated:** [April 3, 2025, 3:26pm UTC](https://forum.microlist.org/t/consult-the-excitation-optical-path-of-tirf/3054 "2025-04-03T15:26:43Z")

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Hello everyone. Recently, I have been building a TIRF microscope based on the objective lens. I found that after adjusting to the TIRF angle, the totally reflected laser converges at the exit of the optical fiber after p…

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## [High background signal in the 647 channel of TIRF](https://forum.microlist.org/t/high-background-signal-in-the-647-channel-of-tirf/3026)

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**Author:** [@SLEEzZ](https://forum.microlist.org/u/SLEEzZ)\
**Replies:** 2\
**Last updated:** [March 13, 2025, 2:49am UTC](https://forum.microlist.org/t/high-background-signal-in-the-647-channel-of-tirf/3026 "2025-03-13T02:49:06Z")

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Hello everyone. Recently, I have been optimizing a TIRF microscope that has not been used for a long time and conducting research related to smFRET. However, I have encountered the problem of a high background in the 647…

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## [Basic microscopy Questions about building](https://forum.microlist.org/t/basic-microscopy-questions-about-building/2431)

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**Author:** [@mediocremicroscoper](https://forum.microlist.org/u/mediocremicroscoper)\
**Replies:** 16\
**Last updated:** [April 26, 2024, 9:33am UTC](https://forum.microlist.org/t/basic-microscopy-questions-about-building/2431 "2024-04-26T09:33:27Z")

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Hello everyone, Ive recently been tasked with building an inverted objective-tirf microscope. Im rather new to this so im learning by doing. I have built the system and aligned the laser, as far as I know. After alignin…

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## [Vertical Noise Pattern in TIRF Images](https://forum.microlist.org/t/vertical-noise-pattern-in-tirf-images/1165)

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**Author:** [@Sneha\_Paul](https://forum.microlist.org/u/Sneha_Paul)\
**Replies:** 4\
**Last updated:** [October 3, 2020, 4:39pm UTC](https://forum.microlist.org/t/vertical-noise-pattern-in-tirf-images/1165 "2020-10-03T16:39:27Z")

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I am using a TIRF microscope coupled with an EMCCD camera to image cells. I get a wierd pattern in the vertical direction which increases with increase in signal, camerga gain, laser intensity. Can anyone help.

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## [TIRF Single Molecule Fluorescence with 488nm - Objective Autofluorescence](https://forum.microlist.org/t/tirf-single-molecule-fluorescence-with-488nm-objective-autofluorescence/854)

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**Author:** [@langevin](https://forum.microlist.org/u/langevin)\
**Replies:** 4\
**Last updated:** [April 21, 2020, 5:47pm UTC](https://forum.microlist.org/t/tirf-single-molecule-fluorescence-with-488nm-objective-autofluorescence/854 "2020-04-21T17:47:02Z")

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Hi everyone, I was wondering if anyone of you has looked into the background generated by autofluorescence of standard (TIRF) objectives. I have set up a TIRF microscope for single molecule imaging with laser lines 488…