Time-Resolved Fluorescence Wiki

how_to_measure_the_instrument_response_function_irf

Anonymous (anonymous@undisclosed.example.com) — 2023-09-07T22:55:03+00:00

2023/09/08 00:54		current
Line 22:		Line 22:
	===== Using samples with ultrafast decay =====		===== Using samples with ultrafast decay =====

-	Some detectors (particularly SPADs) have wavelength dependent timing response. In this case an IRF recorded at the excitation wavelength may not be useful for precise reconvolution. The solution is to acquire the IRF at the fluorescence wavelength, or at least spectrally closer to the fluorescence emission.	+	Some detectors (particularly MPD SPADs) have a wavelength dependent timing response. In this case an IRF recorded at the excitation wavelength may not be useful for precise reconvolution. The solution is to acquire the IRF at the fluorescence wavelength, or at least spectrally closer to the fluorescence emission wavelength.

	==== General recipe ====		==== General recipe ====

align_beam_backreflection

Anonymous (anonymous@undisclosed.example.com) — 2022-08-29T11:17:12+00:00

2022/08/29 13:15		current
Line 12:		Line 12:
	Adding a drop of water on top of the coverslip reduces the back-scattering intensity by an order of magnitude or so, but the basic shape remains the same. Even if one removes everything extra from the beam path, these fringes remain there, they are universal. Note that all those images above are slightly defocused.		Adding a drop of water on top of the coverslip reduces the back-scattering intensity by an order of magnitude or so, but the basic shape remains the same. Even if one removes everything extra from the beam path, these fringes remain there, they are universal. Note that all those images above are slightly defocused.

-	Such images are used for the daily and fundamental alignment of the MicroTime 200. The detailed procedure is described in the user manual of MicroTime 200. Although the MicroTime is extremely robust we suggest to spend 5-10 minutes every day for verifying the quality of the alignment by inspecting the back-reflection pattern. Have a look how this is used also by watching the tutorial [[howto:exchange_dichroic_mt200\|How to exchange the main dichroic of the MicroTime 200.]]	+	Such images are used for daily and fundamental alignment of the MicroTime 200. A detailed procedure is described in the user manual of MicroTime 200. Although MicroTime is extremely robust, we suggest to spend 5-10 minutes every day for verifying the quality of the alignment by inspecting the back-reflection pattern. Have a look how this is used also by watching the tutorial [[howto:exchange_dichroic_mt200\|How to exchange the main dichroic of the MicroTime 200.]]

	The paper by [[ http://rspa.royalsocietypublishing.org/content/253/1274/358\|B. Richards and E. Wolf: Electromagnetic diffraction in optical systems, II. Structure of the image field in an aplanatic system]] shows such 4 lobed patterns for polarized light.		The paper by [[ http://rspa.royalsocietypublishing.org/content/253/1274/358\|B. Richards and E. Wolf: Electromagnetic diffraction in optical systems, II. Structure of the image field in an aplanatic system]] shows such 4 lobed patterns for polarized light.

	Similar figures can be found in this [[https://books.google.cz/books?id=kURdAAAAQBAJ\|classic textbook]].		Similar figures can be found in this [[https://books.google.cz/books?id=kURdAAAAQBAJ\|classic textbook]].

exchange_dichroic_mt200

Anonymous (anonymous@undisclosed.example.com) — 2023-02-17T12:42:54+00:00

2021/06/25 11:19		current
Line 1:		Line 1:
-		+	{{tag>howto MicroTime200 dichroic microscopy}}
-	{{tag>howto MicroTime200 dichroic microscopy MT200}}	+

	====== How to exchange the main dichroic of the MicroTime 200 ======		====== How to exchange the main dichroic of the MicroTime 200 ======

flim-fret_calculation_for_single_exponential_donors

Anonymous (anonymous@undisclosed.example.com) — 2020-05-06T09:38:57+00:00

2020/05/06 11:19		current
Line 85:		Line 85:

	Response: In the image, only pixels with higher photon counts are highlighted. Adapt the intensity maximum to see the highlighted pixels more clearly.		Response: In the image, only pixels with higher photon counts are highlighted. Adapt the intensity maximum to see the highlighted pixels more clearly.
-	{{ :howto:flim-fret-1expd_roi_from_thresh_2_v2.png?400 \|}}	+	{{ :howto:flim-fret-1expd_roi_from_thresh_2_v3.png?400 \|}}

	* In the lifetime fitting panel, keep the default Fitting model "Mono-Exp. Donor".		* In the lifetime fitting panel, keep the default Fitting model "Mono-Exp. Donor".

start

Anonymous (anonymous@undisclosed.example.com) — 2026-02-11T14:19:03+00:00

2024/12/19 16:57		current
Line 8:		Line 8:
	{{topic>howto -FT300 -sample -analysis -draft &nodate&nouser}}		{{topic>howto -FT300 -sample -analysis -draft &nodate&nouser}}

-	===== NovaFLIM / Analysis =====
-
-	{{topic>howto +tutorial +novaflim -draft &nodate&nouser}}

	===== SymPhoTime64 / Analysis =====		===== SymPhoTime64 / Analysis =====

symphotime_tips_and_tricks

Anonymous (anonymous@undisclosed.example.com) — 2020-04-03T14:54:26+00:00

2020/04/03 16:54		current
Line 1:		Line 1:

-	~~TOC~~

	{{tag>Howto Analysis Tutorial SymPhoTime Tips&Tricks}}		{{tag>Howto Analysis Tutorial SymPhoTime Tips&Tricks}}
Line 7:		Line 6:

	====== SymPhoTime 64 Analysis Tips and Tricks ======		====== SymPhoTime 64 Analysis Tips and Tricks ======
		+
		+	~~TOC~~
		+
	===== Summary =====		===== Summary =====

phasor_plot_structure_separation

Anonymous (anonymous@undisclosed.example.com) — 2025-06-16T08:36:34+00:00

2025/06/16 10:36		current
Line 1:		Line 1:
	{{tag> FLIM acquisition Nikon AX Phasor video LSM_upgrade howto}}		{{tag> FLIM acquisition Nikon AX Phasor video LSM_upgrade howto}}

-	====== Phasor-Based Structural Separation Using FLIM ======	+	====== Nikon AX: Phasor-Based Structural Separation Using FLIM ======

	Introduction		Introduction