Advanced Fluorescence and Fluorescence Microscopy - Bewerkingsoverzicht

R0893093: /* Exam 23 January 2026 - Afternoon */

2026-01-25T13:42:53Z

Exam 23 January 2026 - Afternoon

R0893093: /* Examenvragen */

2026-01-23T18:57:12Z

Examenvragen

← Oudere versie		Versie van 23 jan 2026 20:57
Regel 20:		Regel 20:
	#Calculate how long imaging would take for a) 3D widefield imaging and B) 3d dSTORM imaging. For a sample of 120x120 µm and 80µm thick. Give all assumptions you made and justify why. (He mainly wanted to see you thought about everything, Field of view, stage movement, readout time...)		#Calculate how long imaging would take for a) 3D widefield imaging and B) 3d dSTORM imaging. For a sample of 120x120 µm and 80µm thick. Give all assumptions you made and justify why. (He mainly wanted to see you thought about everything, Field of view, stage movement, readout time...)

	A small region is first bleached with a strong laser, which removes fluorescence there. The recovery of fluorescence over time is then measured as unbleached molecules move back into the area. This reveals information about diffusion, binding, and molecular dynamics of proteins or lipids in living cells. ~~(mobile and immobile fraction where replaced by fraction 1 & fraction 2)~~		A small region is first bleached with a strong laser, which removes fluorescence there. The recovery of fluorescence over time is then measured as unbleached molecules move back into the area. This reveals information about diffusion, binding, and molecular dynamics of proteins or lipids in living cells. [Standard FRAP graph was provided]
	[[~~Media:~~FRAP~~.jpg]~~]
	#Which is true? (a)Fraction 1=immobile fraction, Fraction 2=mobile fraction, (b) Fraction 1=mobile fraction, Fraction 2=immobile fraction		#Which is true? (a)Fraction 1=immobile fraction, Fraction 2=mobile fraction, (b) Fraction 1=mobile fraction, Fraction 2=immobile fraction
	#Draw the graph as it would look in case diffusion was slower.		#Draw the graph as it would look in case diffusion was slower.

R0893093: /* Examenvragen */

2026-01-23T18:55:52Z

Examenvragen

← Oudere versie		Versie van 23 jan 2026 20:55
Regel 12:		Regel 12:

	== Examenvragen ==		== Examenvragen ==
			===Exam 23 January 2026 - Afternoon===
			Actine is typically labeled using phalloidin with an organic dye attached. The actin itself has a diameter of 7nm, the combination with the dyes has a diameter of 12nm.
			#Calculate the diffraction limited resolution when imaging using widefield with a 100x magnification, NA=1.49 objective. The emission has a wavelength of 580nm.
			#Can this resolution be achieved using a 2048x2048 detector with pixel sizes of 6.5x6.5µm?
			#This actine is imaged as described in 1a using dSTORM. Described the structural resolution of this imaging? (This was worded very vague and weirdly, the idea was that you should realize that the actin-dye complex is smaller than the 20-30nm limit of STORM, the only way to improve is by improving localisation)
			#How can this dSTORM resolution be improved?
			#Calculate how long imaging would take for a) 3D widefield imaging and B) 3d dSTORM imaging. For a sample of 120x120 µm and 80µm thick. Give all assumptions you made and justify why. (He mainly wanted to see you thought about everything, Field of view, stage movement, readout time...)

			A small region is first bleached with a strong laser, which removes fluorescence there. The recovery of fluorescence over time is then measured as unbleached molecules move back into the area. This reveals information about diffusion, binding, and molecular dynamics of proteins or lipids in living cells. (mobile and immobile fraction where replaced by fraction 1 & fraction 2)
			[[Media:FRAP.jpg]]
			#Which is true? (a)Fraction 1=immobile fraction, Fraction 2=mobile fraction, (b) Fraction 1=mobile fraction, Fraction 2=immobile fraction
			#Draw the graph as it would look in case diffusion was slower.
			#In case you want to image very fast dynamics. Would you bleach a larger or smaller volume?

			A transcription factor binds specific sequences in the nucleus and resides there for 1-5 seconds. What technique would you propose to investigate the time bound vs free. Explain what imaging technique you would use and how you would handle the analysis?

	===Exam 25 January 2025===		===Exam 25 January 2025===

R0893093: /* Vakinformatie */

2026-01-20T08:46:59Z

Vakinformatie

← Oudere versie		Versie van 20 jan 2026 10:46
Regel 7:		Regel 7:
	Boeiend vak, gegeven door Johan Hofkens, Maarten Roeffaers, Elke Debroye en Jelle Hendrix.		Boeiend vak, gegeven door Johan Hofkens, Maarten Roeffaers, Elke Debroye en Jelle Hendrix.
	Vak telt ook soort van (niet verplichte) practicumsessies, waarin je kort ingeleid wordt tot diverse microscopische/fluorescentie-technieken.		Vak telt ook soort van (niet verplichte) practicumsessies, waarin je kort ingeleid wordt tot diverse microscopische/fluorescentie-technieken.
	Mondeling examen dat open boek is, <s>waarbij je alles (inclusief laptop met internet) mag gebruiken.</s> Sinds 2025-2026 mogen enkel afgedrukte bladeren gebruikt worden (zeer groen enzo), door overmatig gebruik van AI.		Mondeling examen dat open boek is, <s>waarbij je alles (inclusief laptop met internet) mag gebruiken.</s> Sinds 2025-2026 mogen enkel afgedrukte bladeren gebruikt worden (zeer groen enzo), door overmatig gebruik van AI. Jelle Hendrix geeft de cursus ook niet meer sinds 2025-'26. Elke is enkel aanwezig bij de eerste les en de presentaties.

	ECTS-fiche: https://onderwijsaanbod.kuleuven.be/syllabi/e/G0G59AE.htm		ECTS-fiche: https://onderwijsaanbod.kuleuven.be/syllabi/e/G0G59AE.htm

R0893093: /* Vakinformatie */

2026-01-13T07:58:07Z

Vakinformatie

← Oudere versie		Versie van 13 jan 2026 09:58
Regel 7:		Regel 7:
	Boeiend vak, gegeven door Johan Hofkens, Maarten Roeffaers, Elke Debroye en Jelle Hendrix.		Boeiend vak, gegeven door Johan Hofkens, Maarten Roeffaers, Elke Debroye en Jelle Hendrix.
	Vak telt ook soort van (niet verplichte) practicumsessies, waarin je kort ingeleid wordt tot diverse microscopische/fluorescentie-technieken.		Vak telt ook soort van (niet verplichte) practicumsessies, waarin je kort ingeleid wordt tot diverse microscopische/fluorescentie-technieken.
	Mondeling examen dat open boek is, waarbij je alles (inclusief laptop met internet) mag gebruiken.		Mondeling examen dat open boek is, <s>waarbij je alles (inclusief laptop met internet) mag gebruiken.</s> Sinds 2025-2026 mogen enkel afgedrukte bladeren gebruikt worden (zeer groen enzo), door overmatig gebruik van AI.

	ECTS-fiche: https://onderwijsaanbod.kuleuven.be/syllabi/e/G0G59AE.htm		ECTS-fiche: https://onderwijsaanbod.kuleuven.be/syllabi/e/G0G59AE.htm

R0854072: /* Exam 25 January 2025 */

2025-01-25T02:06:19Z

Exam 25 January 2025

← Oudere versie		Versie van 25 jan 2025 04:06
Regel 16:		Regel 16:

	Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)		Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)

	Therapeutic monoclonal antibodies (mAbs) are widley used in oncology and immunotherapy, targeting cell surface receptors to induce cytotoxicity responses. Advanced microscopy techniques are critical for understanding the molecular interactions underlying these processes.		Therapeutic monoclonal antibodies (mAbs) are widley used in oncology and immunotherapy, targeting cell surface receptors to induce cytotoxicity responses. Advanced microscopy techniques are critical for understanding the molecular interactions underlying these processes.

R0854072: /* Exam 25 January 2025 */

2025-01-25T02:05:29Z

Exam 25 January 2025

← Oudere versie		Versie van 25 jan 2025 04:05
Regel 22:		Regel 22:

	Roeffaers		Roeffaers
	#* Photoactivatible GFP: Can you use this with A) Confocal B) PALM C) STED, if yes, draw the setup, explain the advantages and disadvantages of this technique for imaging biological samples.(draw the setup, including the laser that activates the PA-GFP! Roeffaers only looks at your drawings so make sure they are good and complete)		# Photoactivatible GFP: Can you use this with A) Confocal B) PALM C) STED, if yes, draw the setup, explain the advantages and disadvantages of this technique for imaging biological samples.(draw the setup, including the laser that activates the PA-GFP! Roeffaers only looks at your drawings so make sure they are good and complete)
	#* Bereken een golfgetal om in een golflengte		# Bereken een golfgetal om in een golflengte

	===Exam 28 January 2022===		===Exam 28 January 2022===

R0854072: /* Exam 25 January 2025 */

2025-01-25T02:05:08Z

Exam 25 January 2025

← Oudere versie		Versie van 25 jan 2025 04:05
Regel 16:		Regel 16:

	Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)		Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)
	#Therapeutic monoclonal antibodies (mAbs) are widley used in oncology and immunotherapy, targeting cell surface receptors to induce cytotoxicity responses. Advanced microscopy techniques are critical for understanding the molecular interactions underlying these processes.		Therapeutic monoclonal antibodies (mAbs) are widley used in oncology and immunotherapy, targeting cell surface receptors to induce cytotoxicity responses. Advanced microscopy techniques are critical for understanding the molecular interactions underlying these processes.

	#1)Design an optical microscopy-based experiment to study how mAbs interact with a specific cell surface receptor (e.g. clustering, cross-linking, or receptor redistribution). In your answer: Identify the micrscopy technique(s) you would use and justify your choice(s). Explain the labeling strategy for both the mAbs and the receptor of interest, considering resolution, specificity, and live-cell imaging requirements. Describe the controls you would include to ensure reliable interpretation of your data.		#Design an optical microscopy-based experiment to study how mAbs interact with a specific cell surface receptor (e.g. clustering, cross-linking, or receptor redistribution). In your answer: Identify the micrscopy technique(s) you would use and justify your choice(s). Explain the labeling strategy for both the mAbs and the receptor of interest, considering resolution, specificity, and live-cell imaging requirements. Describe the controls you would include to ensure reliable interpretation of your data.
			# Imagine that your chosen microscopy method reveals receptor clustering on microvili-like cellular protrusions following mAb RTX binding (as can be seen in the paper at Figure 4A). Propose a hypothesis for how this clustering might influence immune cell recruitment or receptor signaling, and outline an experimental plan to test your hypothesis.
	#2) Imagine that your chosen microscopy method reveals receptor clustering on microvili-like cellular protrusions following mAb RTX binding (as can be seen in the paper at Figure 4A). Propose a hypothesis for how this clustering might influence immune cell recruitment or receptor signaling, and outline an experimental plan to test your hypothesis.

	Roeffaers		Roeffaers

R0854072: /* Exam 25 January 2022 */

2025-01-25T02:04:32Z

Exam 25 January 2022

← Oudere versie		Versie van 25 jan 2025 04:04
Regel 13:		Regel 13:
	== Examenvragen ==		== Examenvragen ==

	===Exam 25 January ~~2022~~===		===Exam 25 January 2025===

	Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)		Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)

R0854072: /* Examenvragen */

2025-01-25T02:04:12Z

Examenvragen

← Oudere versie		Versie van 25 jan 2025 04:04
Regel 12:		Regel 12:

	== Examenvragen ==		== Examenvragen ==

			===Exam 25 January 2022===

			Hofkens (question is based on following paper: https://www.biorxiv.org/content/10.1101/2023.10.09.561472v2.full)
			#Therapeutic monoclonal antibodies (mAbs) are widley used in oncology and immunotherapy, targeting cell surface receptors to induce cytotoxicity responses. Advanced microscopy techniques are critical for understanding the molecular interactions underlying these processes.

			#1)Design an optical microscopy-based experiment to study how mAbs interact with a specific cell surface receptor (e.g. clustering, cross-linking, or receptor redistribution). In your answer: Identify the micrscopy technique(s) you would use and justify your choice(s). Explain the labeling strategy for both the mAbs and the receptor of interest, considering resolution, specificity, and live-cell imaging requirements. Describe the controls you would include to ensure reliable interpretation of your data.

			#2) Imagine that your chosen microscopy method reveals receptor clustering on microvili-like cellular protrusions following mAb RTX binding (as can be seen in the paper at Figure 4A). Propose a hypothesis for how this clustering might influence immune cell recruitment or receptor signaling, and outline an experimental plan to test your hypothesis.

			Roeffaers
			#* Photoactivatible GFP: Can you use this with A) Confocal B) PALM C) STED, if yes, draw the setup, explain the advantages and disadvantages of this technique for imaging biological samples.(draw the setup, including the laser that activates the PA-GFP! Roeffaers only looks at your drawings so make sure they are good and complete)
			#* Bereken een golfgetal om in een golflengte

	===Exam 28 January 2022===		===Exam 28 January 2022===

← Oudere versie		Versie van 25 jan 2026 15:42
Regel 14:		Regel 14:
	===Exam 23 January 2026 - Afternoon===		===Exam 23 January 2026 - Afternoon===
	Actine is typically labeled using phalloidin with an organic dye attached. The actin itself has a diameter of 7nm, the combination with the dyes has a diameter of 12nm.		Actine is typically labeled using phalloidin with an organic dye attached. The actin itself has a diameter of 7nm, the combination with the dyes has a diameter of 12nm.
	#Calculate the diffraction limited resolution when imaging using widefield with a 100x magnification, NA=1.49 objective. The emission has a wavelength of 580nm.		#Calculate the diffraction limited resolution when imaging using widefield with a 100x magnification, NA=1.49 objective. The emission has a wavelength of 580nm. (2p)
	#Can this resolution be achieved using a 2048x2048 detector with pixel sizes of 6.5x6.5µm?		#Can this resolution be achieved using a 2048x2048 detector with pixel sizes of 6.5x6.5µm? (2p)
	#This actine is imaged as described in 1a using dSTORM. Described the structural resolution of this imaging? (This was worded very vague and weirdly, the idea was that you should realize that the actin-dye complex is smaller than the 20-30nm limit of STORM, the only way to improve is by improving localisation)		#This actine is imaged as described in 1a using dSTORM. Described the structural resolution of this imaging? (3p) (This was worded very vague and weirdly, the idea was that you should realize that the actin-dye complex is smaller than the 20-30nm limit of STORM, the only way to improve is by improving localisation)
	#How can this dSTORM resolution be improved?		#How can this dSTORM resolution be improved? (1p)
	#Calculate how long imaging would take for a) 3D widefield imaging and B) 3d dSTORM imaging. For a sample of 120x120 µm and 80µm thick. Give all assumptions you made and justify why. (He mainly wanted to see you thought about everything, Field of view, stage movement, readout time...)		#Calculate how long imaging would take for a) 3D widefield imaging and B) 3d dSTORM imaging. For a sample of 120x120 µm and 80µm thick. Give all assumptions you made and justify why. (2p) (He mainly wanted to see you thought about everything, Field of view, stage movement, readout time...)

	A small region is first bleached with a strong laser, which removes fluorescence there. The recovery of fluorescence over time is then measured as unbleached molecules move back into the area. This reveals information about diffusion, binding, and molecular dynamics of proteins or lipids in living cells. [Standard FRAP graph was provided]		A small region is first bleached with a strong laser, which removes fluorescence there. The recovery of fluorescence over time is then measured as unbleached molecules move back into the area. This reveals information about diffusion, binding, and molecular dynamics of proteins or lipids in living cells. [Standard FRAP graph was provided]

	#Which is true? (a)Fraction 1=immobile fraction, Fraction 2=mobile fraction, (b) Fraction 1=mobile fraction, Fraction 2=immobile fraction		#Which is true? (a)Fraction 1=immobile fraction, Fraction 2=mobile fraction, (b) Fraction 1=mobile fraction, Fraction 2=immobile fraction (2p)
	#Draw the graph as it would look in case diffusion was slower.		#Draw the graph as it would look in case diffusion was slower. (2p)
	#In case you want to image very fast dynamics. Would you bleach a larger or smaller volume?		#In case you want to image very fast dynamics. Would you bleach a larger or smaller volume? (3p)

	A transcription factor binds specific sequences in the nucleus and resides there for 1-5 seconds. What technique would you propose to investigate the time bound vs free. Explain what imaging technique you would use and how you would handle the analysis?		A transcription factor binds specific sequences in the nucleus and resides there for 1-5 seconds. What technique would you propose to investigate the time bound vs free. Explain what imaging technique you would use and how you would handle the analysis? (3p)

	===Exam 25 January 2025===		===Exam 25 January 2025===