Time-Resolved Fluorescence Wiki

diamond_nv_centers

Anonymous (anonymous@undisclosed.example.com) — 2018-04-19T12:11:55+00:00

2018/04/19 14:11		current
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	see also [[wp>Nitrogen-vacancy center]]		see also [[wp>Nitrogen-vacancy center]]

-	{{ howto:model_nv-centers.png\|Figure1: Model of the nitrogene vacancy in the diamond lattice}}	+	[{{ howto:model_nv-centers.png\|Figure1: Model of the nitrogene vacancy in the diamond lattice}}]

	==== Excitation ====		==== Excitation ====

2ffcs

Anonymous (anonymous@undisclosed.example.com) — 2015-11-04T09:41:00+00:00

2014/04/09 22:02		current
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	Correlation analysis that is applied to the fluctuations of the fluorescence intensity. The cross-correlation between the occurrence of events in two different detection volumes is used to introduce an absolute diffusion length into the analysis.		Correlation analysis that is applied to the fluctuations of the fluorescence intensity. The cross-correlation between the occurrence of events in two different detection volumes is used to introduce an absolute diffusion length into the analysis.
		+
		+	see [[glossary:FCS]] for a introduction on FCS.

	===== Experimental Setup =====		===== Experimental Setup =====

align_beam_backreflection

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

2022/08/29 13:15		current
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	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]].

select_the_correct_pinhole_size

Anonymous (anonymous@undisclosed.example.com) — 2015-09-11T15:24:14+00:00

2015/09/11 17:22		current
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	====== How-to select the correct pinhole size ======		====== How-to select the correct pinhole size ======

-	The pinhole size should be slightly bigger than the image of the [[https://en.wikipedia.org/wiki/Airy_disk\|Airy Disc]] at the position of the pinhole:	+	The pinhole size should be slightly bigger than the image of the [[wp>Airy Disc]] at the position of the pinhole:

	$$d_{Airy} = (1.22 * lambda / N.A.) * M$$		$$d_{Airy} = (1.22 * lambda / N.A.) * M$$

calculate_fccs_trace_with_the_grouped_fcs_script

Anonymous (anonymous@undisclosed.example.com) — 2020-04-01T15:05:26+00:00

2015/02/25 15:01		current
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	~~NOTOC~~		~~NOTOC~~

-	====== Calculate Multiple FCCS Traces with the Grouped FCS Script ======	+	====== Calculate FCCS Traces with the Grouped FCS Script ======

	===== Summary =====		===== Summary =====

check_overlap_of_different_color_confocal_volumes

Anonymous (anonymous@undisclosed.example.com) — 2015-02-25T13:55:02+00:00

2014/01/30 17:02		current
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	{{tag>howto confocal_volume}}		{{tag>howto confocal_volume}}
-	====== How to check the overlap of different color confocal volumes ======	+	====== How to Check the Overlap of Different Color Confocal Volumes ======

	Because both lasers are fed through the same fiber, there cannot be any optical beam displacement. According to my records, your system has the latest apochromatic fiber outcoupler, therefore there is practically nothing you can improve: lasers mounted in the [[products:LCU]] are always well overlapped. This is a simple consequence of the [[Products:MicroTime 200]] design.		Because both lasers are fed through the same fiber, there cannot be any optical beam displacement. According to my records, your system has the latest apochromatic fiber outcoupler, therefore there is practically nothing you can improve: lasers mounted in the [[products:LCU]] are always well overlapped. This is a simple consequence of the [[Products:MicroTime 200]] design.

mt200everyday_alignment

Anonymous (anonymous@undisclosed.example.com) — 2023-02-17T12:43:32+00:00

2021/06/30 15:02		current
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	======MT200 Daily Alignment ======		======MT200 Daily Alignment ======
-	{{tag>howto mt200 video tutorial}}	+	{{tag>howto MicroTime200 video tutorial}}

	This procedure is a quick check of the optical hardware and evaluates the overall system performance. It		This procedure is a quick check of the optical hardware and evaluates the overall system performance. It

using_the_anisotropy_image_script

Anonymous (anonymous@undisclosed.example.com) — 2018-04-12T11:44:14+00:00

2018/04/12 12:26		current
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	Note:		Note:
-	The anisotropy values can theoretically spread between -0.5 and 1 for parallel or vertical excitation and emission dipoles respectively. The anisotropy value of 0 is expected in case of very fast rotation or extremely efficient energy transfer processes (e.g. efficient HOMO-FRET). When measuring a randomly aligned ensemble due to photoselection the anisotropy varies with in the limits of -0.2 and 0.4 instead of -0.5 and 1.	+	The anisotropy values can theoretically spread between -0.5 and 1 for perpendicular and parallel excitation and emission dipoles respectively. The anisotropy value of 0 is expected in case of very fast rotation or extremely efficient energy transfer processes (e.g. efficient HOMO-FRET). When measuring a randomly aligned ensemble due to photoselection the anisotropy varies with in the limits of -0.2 and 0.4 instead of -0.5 and 1.

	==== Adapt the script to the image and the system parameters ====		==== Adapt the script to the image and the system parameters ====