Fluorescent Nanodiamonds as Free Radical Sensors in Aging Yeast Cells
van der Laan, Kiran
DOI:
10.33612/diss.112906297
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Publication date: 2020
Link to publication in University of Groningen/UMCG research database
Citation for published version (APA):
van der Laan, K. (2020). Fluorescent Nanodiamonds as Free Radical Sensors in Aging Yeast Cells: a baker’s yeast response to small diamonds with great potential!. Rijksuniversiteit Groningen.
https://doi.org/10.33612/diss.112906297
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Supplementary documents:
Summary / Samenvatting / Acknowledgements / About the
author
Kiran J. van der Laan
Summary
Fluorescent nanodiamonds (FNDs) are a promising new biosensor. Beyond the sparkle, diamonds have a number of unique properties: extreme hardness, great stability, and chemical inertness. Nanodiamonds can be made in very small sizes and a narrow size distribution, and despite this they can still be functionalized on the surface. Nanodiamonds can also host very stable (fluorescent) defects that are protect in the carbon lattice, by which they are protected against bleaching. These defects are utilized for nanoscale sensing since they to e.g. temperature or magnetic fields in their surroundings.
One of these defects is the so-called NV center. In this defect, two neighboring carbon atoms in the lattice are replaced, by a nitrogen atom with a vacancy next to it. This defect gives the diamond interesting properties: not only makes it the diamond fluorescent, but also does it respond to external factors (such as magnetic signals) by giving fluctuations in its fluorescence that can be read out optically. In other words, using the FNDs one can convert a magnetic signal into optically detectable signals. This is the reason that FNDs are proposed as free radical biosensors to detect the magnetic signals of free radicals by optical detection of the diamond’s fluorescence.
Free radicals are produced during natural cell metabolism, but when the natural balance is disturbed they are also associated with diseases and aging. Sensitive methods to detect free radicals are challenging, providing the need for new biosensor such as FNDs. Using the NV center to study free radical activity in side living cells is the main interest in this research. In this work, the necessary preparatory work to enable this application is done by tackling some obstacles and studying the response of cells to FNDs.
In Chapter 1, a general introduction is given into the three main fields that were
involved in this interdisciplinary research. At first, the scientific background is sketched and the relevance of aging research is emphasized. Going further into detail, the free radical theory of aging is explained as well as the applied aging model system. Finally, fluorescent nanodiamonds and how they function as magnetic biosensors are introduced as a new tool to investigate this.
In order to be able to detect free radicals, the FNDs need to be as close as possible to the molecules of interest. This means the FNDs should enter the cell.
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Supplementary documents:
Summary / Samenvatting / Acknowledgements / About the
author
Kiran J. van der Laan
Summary
Fluorescent nanodiamonds (FNDs) are a promising new biosensor. Beyond the sparkle, diamonds have a number of unique properties: extreme hardness, great stability, and chemical inertness. Nanodiamonds can be made in very small sizes and a narrow size distribution, and despite this they can still be functionalized on the surface. Nanodiamonds can also host very stable (fluorescent) defects that are protect in the carbon lattice, by which they are protected against bleaching. These defects are utilized for nanoscale sensing since they to e.g. temperature or magnetic fields in their surroundings.
One of these defects is the so-called NV center. In this defect, two neighboring carbon atoms in the lattice are replaced, by a nitrogen atom with a vacancy next to it. This defect gives the diamond interesting properties: not only makes it the diamond fluorescent, but also does it respond to external factors (such as magnetic signals) by giving fluctuations in its fluorescence that can be read out optically. In other words, using the FNDs one can convert a magnetic signal into optically detectable signals. This is the reason that FNDs are proposed as free radical biosensors to detect the magnetic signals of free radicals by optical detection of the diamond’s fluorescence.
Free radicals are produced during natural cell metabolism, but when the natural balance is disturbed they are also associated with diseases and aging. Sensitive methods to detect free radicals are challenging, providing the need for new biosensor such as FNDs. Using the NV center to study free radical activity in side living cells is the main interest in this research. In this work, the necessary preparatory work to enable this application is done by tackling some obstacles and studying the response of cells to FNDs.
In Chapter 1, a general introduction is given into the three main fields that were
involved in this interdisciplinary research. At first, the scientific background is sketched and the relevance of aging research is emphasized. Going further into detail, the free radical theory of aging is explained as well as the applied aging model system. Finally, fluorescent nanodiamonds and how they function as magnetic biosensors are introduced as a new tool to investigate this.
In order to be able to detect free radicals, the FNDs need to be as close as possible to the molecules of interest. This means the FNDs should enter the cell.
208 209
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Processed on: 21-1-2020 PDF page: 210PDF page: 210PDF page: 210PDF page: 210 Mammalian cells readily ingest the FNDs by themselves, but yeast cells have a
rigid cell wall that prevents this. Therefore, in Chapter 2 we started the quest
for an internalization method that was able to get the FNDs inside the yeast cells without damaging them. By using a chemical transformation mix to temporarily disrupt the cell wall, we managed to do. Moreover, a Fiji protocol was developed to quantify the diamond uptake. Both the uptake protocol and the quantification script are generally applicable for other cell types and fluorescent particles as well.
Chapter 3 confirms the usability of FNDs in the aging model system of choice:
yeast cells. We show that the chemical transformation protocol also works to obtain FND uptake in aged cells. Moreover, we demonstrate that the cells are still aging after this treatment and that the aging curve is not affected by both the uptake protocol and the FNDs. Interestingly, we see that the diamonds are predominantly localized in close proximity of membrane-enclosed structures after.
In Chapter 4 we take the viability measurements a step further and look into
more detail for any non-fatal changes inside the cells, as a response to FND internalization. The stress response of yeast cells is measured by evaluating both metabolic activity, activity of genes involved in different steps and locations of the oxidative stress defense systems and the general free radical activity. Only minimal FND-related stress effects are observed, confirming the excellent biocompatibility. This is of course a crucial finding, to move towards the application of FNDs as biosensors in free radical and aging research.
In this thesis, the focus is on the search of the exact role of free radicals in aging by utilizing the magnetic properties of FNDS. However, there are more interesting applications possible for nanodiamonds in biological systems. Diamond defect properties also include sensing temperature, electric fields or strain and pressure. In Chapter 5 a literature research was done on the
nanodiamond application in cells. We discuss ways to chemically modify the surface of diamonds, and specific issues when dealing with cells, like biocompatibility and how to internalize the diamonds. Applications that are discussed include labeling, sensing, drug delivery, theranostics, antibiotics and tissue engineering. In Chapter 6, the application possibilities for in vivo systems
are discussed. This is especially relevant for nanodiamonds, as their stable fluorescent defects are protected in the lattice, which means that they do not bleach which makes them suitable for long term tracking and sensing in living organisms. Here we discuss different ways to administer nanodiamonds to several model organisms, and how these organisms respond to that. The in vivo applications discussed, involve drug delivery, aiding radiology, labeling and use in cosmetics. Next to that, we also shortly discuss future developments concerning (clinical) applications in humans and how to continue towards the clinic with this.
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Processed on: 21-1-2020 PDF page: 211PDF page: 211PDF page: 211PDF page: 211 Mammalian cells readily ingest the FNDs by themselves, but yeast cells have a
rigid cell wall that prevents this. Therefore, in Chapter 2 we started the quest
for an internalization method that was able to get the FNDs inside the yeast cells without damaging them. By using a chemical transformation mix to temporarily disrupt the cell wall, we managed to do. Moreover, a Fiji protocol was developed to quantify the diamond uptake. Both the uptake protocol and the quantification script are generally applicable for other cell types and fluorescent particles as well.
Chapter 3 confirms the usability of FNDs in the aging model system of choice:
yeast cells. We show that the chemical transformation protocol also works to obtain FND uptake in aged cells. Moreover, we demonstrate that the cells are still aging after this treatment and that the aging curve is not affected by both the uptake protocol and the FNDs. Interestingly, we see that the diamonds are predominantly localized in close proximity of membrane-enclosed structures after.
In Chapter 4 we take the viability measurements a step further and look into
more detail for any non-fatal changes inside the cells, as a response to FND internalization. The stress response of yeast cells is measured by evaluating both metabolic activity, activity of genes involved in different steps and locations of the oxidative stress defense systems and the general free radical activity. Only minimal FND-related stress effects are observed, confirming the excellent biocompatibility. This is of course a crucial finding, to move towards the application of FNDs as biosensors in free radical and aging research.
In this thesis, the focus is on the search of the exact role of free radicals in aging by utilizing the magnetic properties of FNDS. However, there are more interesting applications possible for nanodiamonds in biological systems. Diamond defect properties also include sensing temperature, electric fields or strain and pressure. In Chapter 5 a literature research was done on the
nanodiamond application in cells. We discuss ways to chemically modify the surface of diamonds, and specific issues when dealing with cells, like biocompatibility and how to internalize the diamonds. Applications that are discussed include labeling, sensing, drug delivery, theranostics, antibiotics and tissue engineering. In Chapter 6, the application possibilities for in vivo systems
are discussed. This is especially relevant for nanodiamonds, as their stable fluorescent defects are protected in the lattice, which means that they do not bleach which makes them suitable for long term tracking and sensing in living organisms. Here we discuss different ways to administer nanodiamonds to several model organisms, and how these organisms respond to that. The in vivo applications discussed, involve drug delivery, aiding radiology, labeling and use in cosmetics. Next to that, we also shortly discuss future developments concerning (clinical) applications in humans and how to continue towards the clinic with this.
210 211
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Samenvatting
Fluorescerende nanodiamanten (FNDs) zijn een veelbelovende nieuwe biosensor. Naast de schittering, hebben diamanten een aantal unieke eigenschappen: extreme hardheid, grote stabiliteit en chemische inertie. Nanodiamanten kunnen worden gemaakt in hele kleine maten en met een smalle grootteverdeling. Ondanks het kleine formaat, kan het oppervlakte van de diamanten wel gefunctionaliseerd worden. Daarnaast kunnen nanodiamanten bepaalde defecten hebben, die in het koolstofrooster zitten, en daardoor beschermd zijn tegen verbleken. Deze defecten worden gebruikt voor sensors op nanoformaat, omdat ze reageren op bijvoorbeeld de temperatuur en magnetische velden in de omgeving.
Één van deze defecten is het zogenoemde nitrogen vacancy (NV) center, waarbij twee naburige koolstofatomen zijn vervangen door een stikstofatoom en een lege plek daarnaast. Dit NV-center geeft de diamant interessante eigenschappen: het NV center is fluorescent en reageert bovendien op externe factoren in de omgeving (zoals magnetische signalen) door een fluctuatie in de fluorescentie te geven. Met andere woorden, de FNDs kunnen worden gebruikt om magnetische signalen om te zetten in optisch detecteerbare signalen. En dat is dus precies waarom FNDs als vrije radicalen biosensoren kunnen functioneren: men kan de magnetische signalen van vrije radicalen detecteren, door de fluorescentie van de diamant uit te lezen.
Vrije radicalen worden geproduceerd tijdens het natuurlijke metabolisme van de cel, maar wanneer deze natuurlijke balans wordt verstoord kunnen er problemen ontstaan zoals ziektes en veroudering. Gevoelige methoden om vrije radicalen te meten zijn er op dit moment niet, wat het belang van onze FNDs als nieuwe biosensor versterkt. Het gebruik van diamanten met NV-centers met als doel de activiteit van vrije radicalen te bestuderen in levende cellen, is de focus van dit onderzoek. In dit proefschrift, het nodige voorwerk is gedaan om de toepassing mogelijk te maken door een aantal obstakels te overwinnen en door de reactie van cellen op diamanten te bestuderen.
In Hoofdstuk 1, wordt een algemene introductie gegeven van de drie gebieden
die betrokken zijn in dit interdisciplinaire onderzoek. Als eerste wordt de wetenschappelijke achtergrond geschetst en tevens de relevantie van
onderzoek naar verouderen. Daarna meer in detail, wordt de theorie over de rol van vrije radicalen tijdens veroudering uitgelegd, alsmede het toegepaste verouderingsmodel. Tot slot wordt beschreven hoe fluorescerende nanodiamanten werken en hoe zij als magnetische biosensoren kunnen bijdragen aan onderzoek naar veroudering.
Om vrije radicalen in een cel te kunnen meten, moeten de FNDs daar zo dichtbij mogelijk zijn. Dit betekend dat de FNDs moeten worden opgenomen door de cel. Zoogdiercellen nemen FNDs van zichzelf op, maar gistcellen hebben een rigide celwand die dat voorkomt. Daarom gaan we in Hoofdstuk 2 op zoek naar
een manier om de diamanten in de cel te brengen, zonder de gistcellen te beschadigen. Dit is ons gelukt door een mix van chemicaliën te gebruiken om de celwand tijdelijk te verbreken. Bovendien hebben we een Fiji script ontwikkeld om de diamant opname te kwantificeren. Zowel het opname protocol als het kwantificatie script zijn algemeen toepasbaar voor andere cel typen en nanodeeltjes.
Hoofdstuk 3 bevestigt de bruikbaarheid van FNDs in het modelsysteem voor
veroudering dat wordt gebruikt in dit onderzoek: gistcellen. We laten zien dat diamant opname met behulp van de chemische transformatie mix ook succesvol is in al verouderde cellen. Daarnaast demonstreren we dat de cellen nog steeds verouderen na deze behandeling, en dat de snelheid van functionele achteruitgang onveranderd blijft. We observeerden bovendien dat de diamanten zich na opname voornamelijk in de buurt van door membraan omsloten cel organellen bevinden.
In Hoofdstuk 4 gaan we een stap verder en kijken we nog meer in detail naar
non-fatale veranderingen in de cellen, als reactie op FND opname. De stressreactie van gistcellen wordt gemeten door het bestuderen van metabolische activiteit, activiteit van genen betrokken bij verdediging tegen oxidatieve stress, en algemene, totale activiteit van vrije radicalen. De uitstekende biocompatibiliteit wordt bevestigd, omdat slechts minimale FND-gerelateerde stress effecten worden gevonden. Dit is natuurlijk een cruciale bevinding om door te gaan naar de volgende stap, het daadwerkelijk toepassen van FNDs als biosensoren in onderzoek naar vrije radicalen en veroudering.
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Samenvatting
Fluorescerende nanodiamanten (FNDs) zijn een veelbelovende nieuwe biosensor. Naast de schittering, hebben diamanten een aantal unieke eigenschappen: extreme hardheid, grote stabiliteit en chemische inertie. Nanodiamanten kunnen worden gemaakt in hele kleine maten en met een smalle grootteverdeling. Ondanks het kleine formaat, kan het oppervlakte van de diamanten wel gefunctionaliseerd worden. Daarnaast kunnen nanodiamanten bepaalde defecten hebben, die in het koolstofrooster zitten, en daardoor beschermd zijn tegen verbleken. Deze defecten worden gebruikt voor sensors op nanoformaat, omdat ze reageren op bijvoorbeeld de temperatuur en magnetische velden in de omgeving.
Één van deze defecten is het zogenoemde nitrogen vacancy (NV) center, waarbij twee naburige koolstofatomen zijn vervangen door een stikstofatoom en een lege plek daarnaast. Dit NV-center geeft de diamant interessante eigenschappen: het NV center is fluorescent en reageert bovendien op externe factoren in de omgeving (zoals magnetische signalen) door een fluctuatie in de fluorescentie te geven. Met andere woorden, de FNDs kunnen worden gebruikt om magnetische signalen om te zetten in optisch detecteerbare signalen. En dat is dus precies waarom FNDs als vrije radicalen biosensoren kunnen functioneren: men kan de magnetische signalen van vrije radicalen detecteren, door de fluorescentie van de diamant uit te lezen.
Vrije radicalen worden geproduceerd tijdens het natuurlijke metabolisme van de cel, maar wanneer deze natuurlijke balans wordt verstoord kunnen er problemen ontstaan zoals ziektes en veroudering. Gevoelige methoden om vrije radicalen te meten zijn er op dit moment niet, wat het belang van onze FNDs als nieuwe biosensor versterkt. Het gebruik van diamanten met NV-centers met als doel de activiteit van vrije radicalen te bestuderen in levende cellen, is de focus van dit onderzoek. In dit proefschrift, het nodige voorwerk is gedaan om de toepassing mogelijk te maken door een aantal obstakels te overwinnen en door de reactie van cellen op diamanten te bestuderen.
In Hoofdstuk 1, wordt een algemene introductie gegeven van de drie gebieden
die betrokken zijn in dit interdisciplinaire onderzoek. Als eerste wordt de wetenschappelijke achtergrond geschetst en tevens de relevantie van
onderzoek naar verouderen. Daarna meer in detail, wordt de theorie over de rol van vrije radicalen tijdens veroudering uitgelegd, alsmede het toegepaste verouderingsmodel. Tot slot wordt beschreven hoe fluorescerende nanodiamanten werken en hoe zij als magnetische biosensoren kunnen bijdragen aan onderzoek naar veroudering.
Om vrije radicalen in een cel te kunnen meten, moeten de FNDs daar zo dichtbij mogelijk zijn. Dit betekend dat de FNDs moeten worden opgenomen door de cel. Zoogdiercellen nemen FNDs van zichzelf op, maar gistcellen hebben een rigide celwand die dat voorkomt. Daarom gaan we in Hoofdstuk 2 op zoek naar
een manier om de diamanten in de cel te brengen, zonder de gistcellen te beschadigen. Dit is ons gelukt door een mix van chemicaliën te gebruiken om de celwand tijdelijk te verbreken. Bovendien hebben we een Fiji script ontwikkeld om de diamant opname te kwantificeren. Zowel het opname protocol als het kwantificatie script zijn algemeen toepasbaar voor andere cel typen en nanodeeltjes.
Hoofdstuk 3 bevestigt de bruikbaarheid van FNDs in het modelsysteem voor
veroudering dat wordt gebruikt in dit onderzoek: gistcellen. We laten zien dat diamant opname met behulp van de chemische transformatie mix ook succesvol is in al verouderde cellen. Daarnaast demonstreren we dat de cellen nog steeds verouderen na deze behandeling, en dat de snelheid van functionele achteruitgang onveranderd blijft. We observeerden bovendien dat de diamanten zich na opname voornamelijk in de buurt van door membraan omsloten cel organellen bevinden.
In Hoofdstuk 4 gaan we een stap verder en kijken we nog meer in detail naar
non-fatale veranderingen in de cellen, als reactie op FND opname. De stressreactie van gistcellen wordt gemeten door het bestuderen van metabolische activiteit, activiteit van genen betrokken bij verdediging tegen oxidatieve stress, en algemene, totale activiteit van vrije radicalen. De uitstekende biocompatibiliteit wordt bevestigd, omdat slechts minimale FND-gerelateerde stress effecten worden gevonden. Dit is natuurlijk een cruciale bevinding om door te gaan naar de volgende stap, het daadwerkelijk toepassen van FNDs als biosensoren in onderzoek naar vrije radicalen en veroudering.
212 213
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in veroudering, als toepassing om de magnetische eigenschappen van FNDs voor te gebruiken. Echter, er zijn meer interessante toepassingen te bedenken voor nanodiamanten in biologische systemen. Diamantdefecten kunnen bijvoorbeeld temperatuur, elektrische velden of stroom en druk waarnemen. In Hoofdstuk 5
is een literatuuronderzoek uitgevoerd naar de verschillende toepassingen van nanodiamanten in cellen. We bespreken manieren om het oppervlak van de diamanten chemisch te modificeren en ook specifieke zaken die spelen bij het werken met cellen: zoals de biocompatibiliteit en manieren om diamanten te internaliseren. De toepassingen die hier worden besproken zijn onder andere hulp bij medicijn afgifte, antibiotica en weefsel regeneratie en het gebruik van diamanten als labels. In Hoofdstuk 6 worden de toepasmogelijkheden voor in
vivo systemen besproken. Dit is in het bijzonder relevant voor nanodiamanten,
omdat fluorescente defecten beschermd liggen in het koolstofrooster waardoor ze niet verbleken en dus geschikt zijn voor lange termijn metingen in levende organismen. Hier bespreken we verschillende manieren om diamanten toe te dienen bij verschillende organismen, en hoe deze organismen hierop reageren. De in vivo toepassingen die we bespreken zijn medicijn afgifte, ondersteuning radiologie, gebruik in cosmetica en ook weer het gebruik van diamanten als labels. Tevens bespreken we kort de toekomstige ontwikkelingen met betrekking tot (klinische) toepassingen in de mens en hoe verder te gaan om deze naar de kliniek te brengen.
Acknowledgements
Wow, you have made it to the most important part of this book! Of course I could not have done all of this all by myself, so there is a lot of people to thank. First of all, Romana: thank you for giving me the opportunity to start my PhD research here in Groningen. I had not heard of FNDs before I came here, so I am truly thankful that you believed in me and gave me the chance to start with this project. Although I did have some doubts myself from time to time, you never showed any doubts about me or the project itself. I want to thank you as well for cooperating and thinking along with all my plans for the future. I am very impressed by how you handle your professional career, especially in combination with your rich personal life! I wish you, Daniel and the kids a happy life in Groningen for now, and hopefully soon in another city and country! Then the rest of the diamond people, who all have played a big part in my time as being a PhD candidate! Simon, who introduced me in the group and thus actually got me the position. Next to our shared memories of Nuenen and Nijmegen, we could talk about everything, ranging from food to finances to nonsense. It has been a lot of fun to be office mates during my first two years, also together with the other roomies: Aryan, my dear yeast companion and crazy cat lady, I could always count on your help for experiments or drawings and later on cat sitting as well. Felipe, the cookie supplying engineer, we have actually spend almost our whole PhD time together and I am happy to be your paranymph. At last Linyan arrived in the office, with her surprising sense of humour! The office has always been a good spot with a nice atmosphere, good work-chat balance and an overload of cookies! I am also very happy and honored to have you by my side on this special day, Aryan and Felipe.
Actually, in the whole research group we have always had this nice atmosphere with good discussions and also fun activities! This accounts for the people that have left already (Mayeul and Charles), and people that are currently still in the group: Thamir and Yuchen (I really enjoyed our time in Dubrovnik, it was the beset conference visit during my PhD), Yori, Alina (you remind me so much of myself, don’t forget to take it easy from time to time), Viraj, Claudia, Rokshana, Citra, Thea, Neda, Kaiqi, Ari, Sandeep, Aldona (thank you for being my
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in veroudering, als toepassing om de magnetische eigenschappen van FNDs voor te gebruiken. Echter, er zijn meer interessante toepassingen te bedenken voor nanodiamanten in biologische systemen. Diamantdefecten kunnen bijvoorbeeld temperatuur, elektrische velden of stroom en druk waarnemen. In Hoofdstuk 5
is een literatuuronderzoek uitgevoerd naar de verschillende toepassingen van nanodiamanten in cellen. We bespreken manieren om het oppervlak van de diamanten chemisch te modificeren en ook specifieke zaken die spelen bij het werken met cellen: zoals de biocompatibiliteit en manieren om diamanten te internaliseren. De toepassingen die hier worden besproken zijn onder andere hulp bij medicijn afgifte, antibiotica en weefsel regeneratie en het gebruik van diamanten als labels. In Hoofdstuk 6 worden de toepasmogelijkheden voor in
vivo systemen besproken. Dit is in het bijzonder relevant voor nanodiamanten,
omdat fluorescente defecten beschermd liggen in het koolstofrooster waardoor ze niet verbleken en dus geschikt zijn voor lange termijn metingen in levende organismen. Hier bespreken we verschillende manieren om diamanten toe te dienen bij verschillende organismen, en hoe deze organismen hierop reageren. De in vivo toepassingen die we bespreken zijn medicijn afgifte, ondersteuning radiologie, gebruik in cosmetica en ook weer het gebruik van diamanten als labels. Tevens bespreken we kort de toekomstige ontwikkelingen met betrekking tot (klinische) toepassingen in de mens en hoe verder te gaan om deze naar de kliniek te brengen.
Acknowledgements
Wow, you have made it to the most important part of this book! Of course I could not have done all of this all by myself, so there is a lot of people to thank. First of all, Romana: thank you for giving me the opportunity to start my PhD research here in Groningen. I had not heard of FNDs before I came here, so I am truly thankful that you believed in me and gave me the chance to start with this project. Although I did have some doubts myself from time to time, you never showed any doubts about me or the project itself. I want to thank you as well for cooperating and thinking along with all my plans for the future. I am very impressed by how you handle your professional career, especially in combination with your rich personal life! I wish you, Daniel and the kids a happy life in Groningen for now, and hopefully soon in another city and country! Then the rest of the diamond people, who all have played a big part in my time as being a PhD candidate! Simon, who introduced me in the group and thus actually got me the position. Next to our shared memories of Nuenen and Nijmegen, we could talk about everything, ranging from food to finances to nonsense. It has been a lot of fun to be office mates during my first two years, also together with the other roomies: Aryan, my dear yeast companion and crazy cat lady, I could always count on your help for experiments or drawings and later on cat sitting as well. Felipe, the cookie supplying engineer, we have actually spend almost our whole PhD time together and I am happy to be your paranymph. At last Linyan arrived in the office, with her surprising sense of humour! The office has always been a good spot with a nice atmosphere, good work-chat balance and an overload of cookies! I am also very happy and honored to have you by my side on this special day, Aryan and Felipe.
Actually, in the whole research group we have always had this nice atmosphere with good discussions and also fun activities! This accounts for the people that have left already (Mayeul and Charles), and people that are currently still in the group: Thamir and Yuchen (I really enjoyed our time in Dubrovnik, it was the beset conference visit during my PhD), Yori, Alina (you remind me so much of myself, don’t forget to take it easy from time to time), Viraj, Claudia, Rokshana, Citra, Thea, Neda, Kaiqi, Ari, Sandeep, Aldona (thank you for being my
214 215
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but I have loved how it has always felt like a close group! With our pizza lunches, drinks and diamond day; I have enjoyed being part of this team.
I would also like to thank all other colleagues from the Biomedical Engineering department for always being helpful and friendly! To all other PhD candidates (Yuanfeng, Lu, Gwenda, Guangyue, Colin, Laís, Xiaoxiang, Hongpin, Valentina, Torben, Jeroen, Liang, Can, Yanyan, Huaiying, Abigail, Damla, Hao): hold on, you can all make it! Thanks to all the technicians and the secretary as well (Ina, Gesinda, Melissa, Jelly, René, Willy, Betsy, Corien, Ed, Marja, Minie) for all the help and nice chats in the lab and during the coffee breaks.
Other Groningen friends that I want to thank here include my dear fellow Gopher board members; although it has been challenging to combine these activities with the PhD research, I had a lot of fun with all of you! And also Inge; we met each other at the beginning of our PhDs and stayed in contact over the years to discuss our PhD issues. I have valued these discussions a lot and have also enjoyed our coffee and dinner dates, as well as our trip to Copenhagen! It has been a challenge to keep up with friends from the other side of the country as well. But every time I managed to get the time and energy, I enjoyed our outings very much and it has always felt like as it was as yesterday. Friends from Nijmegen (Sonia, Evelyn, Kim, Ilse and Vera) that are now spread all over the country: it has been fun to meet up in different cities, and it is nice to see all of us ending up in different places. Friends from Eindhoven, but now all living in Utrecht: it is impressive how good the connection still is due to our shared time spent in high school, although in some years we only meet during the most fun time of the year (Alaaf!).
Then my dearest friends, Costa Rossa, Jit en Im. Ik heb zoveel steun aan jullie gehad, door dagelijks lief en leed te delen. Ik vind het soms moeilijk dat ik zo ver bij jullie vandaan woon, maar ook juist zo fijn dat we alsnog zo betrokken zijn bij elkaars leven! Ik ben onwijs trots op jullie beiden, op de keuzes die jullie maken en op de overwinningen die jullie behalen. Slurpen voor het leven!
Lieve opa en oma, lieve opa Jan en oma Diny: vroeger zaten we drie uur in de auto om op bezoek te gaan, nu stap ik op de fiets om even langs te komen. Wat is het bijzonder om nu zo dicht bij jullie in Groningen te wonen.
Ook de hele familie van Kalker wil ik graag bedanken voor de gezellige weekenden en de vele bezoekjes aan Groningen!
Dan mijn lieve familie: lieve pap en mam, lieve broer en zus. Wat is het fijn dat jullie er zijn! Ik heb me de laatste jaren steeds meer beseft hoe gelukkig ik mag zijn, met jullie allemaal hier relatief in de buurt, met zulke fijne herinneringen. Lieve Ivar en Jitta, wat mooi om te zien dat we allemaal onze eigen kant opgaan en goed terecht komen. Ook dankzij jullie, lieve pap en mam. Jullie hebben ons altijd de mogelijkheden gegeven, maar ook de vrijheid om onze eigen keuzes te maken. Hierdoor heb ik zoveel kansen gekregen en kunnen pakken, wat misschien niet altijd makkelijk is geweest, maar dit heeft me zeker geholpen de eigenwijze en zelfstandige ik te worden die ik nu ben! Ik kan alleen maar hopen ooit ook zo’n liefdevolle en versterkende ouder te mogen zijn!
Allerliefste Nick, nu 8 jaar geleden ontmoette ik jou in Nijmegen. Ik ben zo gelukkig en dankbaar dat we samen naar Groningen zijn gekomen, om hier samen ons leven verder op te bouwen. Ik heb vaak aan mezelf getwijfeld, of ik het wel zou halen, en of ik het allemaal wel goed genoeg deed ... maar gelukkig was jij altijd daar om me te remmen waar nodig, om me te troosten als het te laat was, en om nooit je geduld te verliezen met mijn soms eigenwijze streken. Wat ben ik trots op ons mooie nieuwe huisje en ons fijne leventje samen. Ik ben heel benieuwd naar alle mooie dingen die ons nog te wachten staan in de toekomst! Gek op je!
539807-L-bw-Laan 539807-L-bw-Laan 539807-L-bw-Laan 539807-L-bw-Laan Processed on: 21-1-2020 Processed on: 21-1-2020 Processed on: 21-1-2020
Processed on: 21-1-2020 PDF page: 217PDF page: 217PDF page: 217PDF page: 217 copromotor) Yue and Runrun. Over the years the group has grown immensely,
but I have loved how it has always felt like a close group! With our pizza lunches, drinks and diamond day; I have enjoyed being part of this team.
I would also like to thank all other colleagues from the Biomedical Engineering department for always being helpful and friendly! To all other PhD candidates (Yuanfeng, Lu, Gwenda, Guangyue, Colin, Laís, Xiaoxiang, Hongpin, Valentina, Torben, Jeroen, Liang, Can, Yanyan, Huaiying, Abigail, Damla, Hao): hold on, you can all make it! Thanks to all the technicians and the secretary as well (Ina, Gesinda, Melissa, Jelly, René, Willy, Betsy, Corien, Ed, Marja, Minie) for all the help and nice chats in the lab and during the coffee breaks.
Other Groningen friends that I want to thank here include my dear fellow Gopher board members; although it has been challenging to combine these activities with the PhD research, I had a lot of fun with all of you! And also Inge; we met each other at the beginning of our PhDs and stayed in contact over the years to discuss our PhD issues. I have valued these discussions a lot and have also enjoyed our coffee and dinner dates, as well as our trip to Copenhagen! It has been a challenge to keep up with friends from the other side of the country as well. But every time I managed to get the time and energy, I enjoyed our outings very much and it has always felt like as it was as yesterday. Friends from Nijmegen (Sonia, Evelyn, Kim, Ilse and Vera) that are now spread all over the country: it has been fun to meet up in different cities, and it is nice to see all of us ending up in different places. Friends from Eindhoven, but now all living in Utrecht: it is impressive how good the connection still is due to our shared time spent in high school, although in some years we only meet during the most fun time of the year (Alaaf!).
Then my dearest friends, Costa Rossa, Jit en Im. Ik heb zoveel steun aan jullie gehad, door dagelijks lief en leed te delen. Ik vind het soms moeilijk dat ik zo ver bij jullie vandaan woon, maar ook juist zo fijn dat we alsnog zo betrokken zijn bij elkaars leven! Ik ben onwijs trots op jullie beiden, op de keuzes die jullie maken en op de overwinningen die jullie behalen. Slurpen voor het leven!
Lieve opa en oma, lieve opa Jan en oma Diny: vroeger zaten we drie uur in de auto om op bezoek te gaan, nu stap ik op de fiets om even langs te komen. Wat is het bijzonder om nu zo dicht bij jullie in Groningen te wonen.
Ook de hele familie van Kalker wil ik graag bedanken voor de gezellige weekenden en de vele bezoekjes aan Groningen!
Dan mijn lieve familie: lieve pap en mam, lieve broer en zus. Wat is het fijn dat jullie er zijn! Ik heb me de laatste jaren steeds meer beseft hoe gelukkig ik mag zijn, met jullie allemaal hier relatief in de buurt, met zulke fijne herinneringen. Lieve Ivar en Jitta, wat mooi om te zien dat we allemaal onze eigen kant opgaan en goed terecht komen. Ook dankzij jullie, lieve pap en mam. Jullie hebben ons altijd de mogelijkheden gegeven, maar ook de vrijheid om onze eigen keuzes te maken. Hierdoor heb ik zoveel kansen gekregen en kunnen pakken, wat misschien niet altijd makkelijk is geweest, maar dit heeft me zeker geholpen de eigenwijze en zelfstandige ik te worden die ik nu ben! Ik kan alleen maar hopen ooit ook zo’n liefdevolle en versterkende ouder te mogen zijn!
Allerliefste Nick, nu 8 jaar geleden ontmoette ik jou in Nijmegen. Ik ben zo gelukkig en dankbaar dat we samen naar Groningen zijn gekomen, om hier samen ons leven verder op te bouwen. Ik heb vaak aan mezelf getwijfeld, of ik het wel zou halen, en of ik het allemaal wel goed genoeg deed ... maar gelukkig was jij altijd daar om me te remmen waar nodig, om me te troosten als het te laat was, en om nooit je geduld te verliezen met mijn soms eigenwijze streken. Wat ben ik trots op ons mooie nieuwe huisje en ons fijne leventje samen. Ik ben heel benieuwd naar alle mooie dingen die ons nog te wachten staan in de toekomst! Gek op je!
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About the author
Kiran van der Laan was born on the 7th of
September 1991 in Eindhoven, the Netherlands. After graduating from the Lorentz Casimir Lyceum in Eindhoven in 2009, she started her bachelor in Medical Biology at the Radboud University in Nijmegen. During her time in Nijmegen, she completed a research internship at the Obstetrics and Gynecology department of the Radboud University Medical Center. Under supervision of dr. Liliana Ramos, she investigated different protocols to optimize decondensation of
sperm cells for quality investigations. In 2012, at the end of the bachelor, Kiran started a secretary position in the study association for a year, next to assisting several freshman courses of the study Biology. Next, she continued with the research master Medical Biology and after some courses she moved to Spain for her master research internship. During her internship in Cordoba, south of Spain, she studied the functional effects as a result of increased hormone expressions in several human endocrine tumor cell lines. The second master research internship was done at the Human Genetics department of the Radboud UMC, where she was working with fruit flies as genetic models to study the role of the FOXP gene which is involved in neurodevelopment.
Following her master graduation in February 2016, Kiran started her PhD in Groningen at the Biomedical Engineering department of the University Medical Center Groningen. She joined the Bioanalysis Research Group, led by prof. dr. Romana Schirghagl. In this project, she focused on the application of Fluorescent Nanodiamonds in the search for causes of cellular ageing. For this purpose, she set up the research direction using the Fluorescent Nanodiamonds in another cell model system used in ageing research: yeast cells. Next to her research, Kiran joined the board of the Groningen Organization for PhD Education and Recreation (GOPHER) as a treasurer from November 2016-November 2017. During her PhD, she presented at several international conferences and she received several awards. In February 2017 she received a personal scholarship
from the Jan Cornelis de Cock foundation, to investigate the oxidative stress response of yeast cells after FND internalization. In March 2017 she received a junior investigator grant for active participation in the Gordon Research Conference on Oxidative Stress & Disease. She also won a travel grant to participate in the first European PhD and Postdoc conference (ENABLE) in July 2017. She is currently applying for a Project Manager or Medical Writer position in Groningen.
Bibliography
2017 van der Laan KJ, Hemelaar SR, Hinterding SR, Koot MV, Ellermann E,
Perona-Martinez FP, Roig D, Hommelet S, Novarina D, Takahashi H, Chang M, Schirhagl R. Generally Applicable Transformation Protocols for Fluorescent Nanodiamond Internalization into Cells. Sci. Rep. 7:
5862 (2017).
2018 Hemelaar SR, Saspaanithy B, L’Hommelet SRM, Perona Martinez FP,
van der Laan KJ, Schirhagl R. The Response of Hela Cells to Fluorescent
NanoDiamond Uptake. Sensors 18: e355 (2018).
Van der Laan K, Hasani M, Zheng T, Schirhagl R. Nanodiamonds for In Vivo Applications. Small 14: e1703838 (2018).
Van der Laan KJ, Chipaux M, Hemelaar SR, Hasani M, Zheng T,
Schirhagl R. Nanodiamonds and Their Applications in Cells. Small 14:
e1704263 (2018).
Van der Laan KJ, Naulleau J, Damle VG, Sigaeva A, Jamot N,
Perona-Martinez FP, Chipaux M, Schirhagl R. Toward Using Fluorescent Nanodiamonds To Study Chronological Aging in Saccharomyces cerevisiae. Anal Chem 90: 13506-13513 (2018).
2019 Castells-Nobau A, Eidhof I, Fenckova M, Brenman-Suttner DB, Scheffer-de Gooyert JM, Christine S, Schellevis RL, van der Laan K, Quentin C,
van Ninhuijs L, Hofmann F, Ejsmont R, Fisher SE, Kramer JM, Sigrist SJ, Simon AF, Schenck A. Conserved regulation of neurodevelopmental
539807-L-bw-Laan 539807-L-bw-Laan 539807-L-bw-Laan 539807-L-bw-Laan Processed on: 21-1-2020 Processed on: 21-1-2020 Processed on: 21-1-2020
Processed on: 21-1-2020 PDF page: 219PDF page: 219PDF page: 219PDF page: 219
About the author
Kiran van der Laan was born on the 7th of
September 1991 in Eindhoven, the Netherlands. After graduating from the Lorentz Casimir Lyceum in Eindhoven in 2009, she started her bachelor in Medical Biology at the Radboud University in Nijmegen. During her time in Nijmegen, she completed a research internship at the Obstetrics and Gynecology department of the Radboud University Medical Center. Under supervision of dr. Liliana Ramos, she investigated different protocols to optimize decondensation of
sperm cells for quality investigations. In 2012, at the end of the bachelor, Kiran started a secretary position in the study association for a year, next to assisting several freshman courses of the study Biology. Next, she continued with the research master Medical Biology and after some courses she moved to Spain for her master research internship. During her internship in Cordoba, south of Spain, she studied the functional effects as a result of increased hormone expressions in several human endocrine tumor cell lines. The second master research internship was done at the Human Genetics department of the Radboud UMC, where she was working with fruit flies as genetic models to study the role of the FOXP gene which is involved in neurodevelopment.
Following her master graduation in February 2016, Kiran started her PhD in Groningen at the Biomedical Engineering department of the University Medical Center Groningen. She joined the Bioanalysis Research Group, led by prof. dr. Romana Schirghagl. In this project, she focused on the application of Fluorescent Nanodiamonds in the search for causes of cellular ageing. For this purpose, she set up the research direction using the Fluorescent Nanodiamonds in another cell model system used in ageing research: yeast cells. Next to her research, Kiran joined the board of the Groningen Organization for PhD Education and Recreation (GOPHER) as a treasurer from November 2016-November 2017. During her PhD, she presented at several international conferences and she received several awards. In February 2017 she received a personal scholarship
from the Jan Cornelis de Cock foundation, to investigate the oxidative stress response of yeast cells after FND internalization. In March 2017 she received a junior investigator grant for active participation in the Gordon Research Conference on Oxidative Stress & Disease. She also won a travel grant to participate in the first European PhD and Postdoc conference (ENABLE) in July 2017. She is currently applying for a Project Manager or Medical Writer position in Groningen.
Bibliography
2017 van der Laan KJ, Hemelaar SR, Hinterding SR, Koot MV, Ellermann E,
Perona-Martinez FP, Roig D, Hommelet S, Novarina D, Takahashi H, Chang M, Schirhagl R. Generally Applicable Transformation Protocols for Fluorescent Nanodiamond Internalization into Cells. Sci. Rep. 7:
5862 (2017).
2018 Hemelaar SR, Saspaanithy B, L’Hommelet SRM, Perona Martinez FP,
van der Laan KJ, Schirhagl R. The Response of Hela Cells to Fluorescent
NanoDiamond Uptake. Sensors 18: e355 (2018).
Van der Laan K, Hasani M, Zheng T, Schirhagl R. Nanodiamonds for In Vivo Applications. Small 14: e1703838 (2018).
Van der Laan KJ, Chipaux M, Hemelaar SR, Hasani M, Zheng T,
Schirhagl R. Nanodiamonds and Their Applications in Cells. Small 14:
e1704263 (2018).
Van der Laan KJ, Naulleau J, Damle VG, Sigaeva A, Jamot N,
Perona-Martinez FP, Chipaux M, Schirhagl R. Toward Using Fluorescent Nanodiamonds To Study Chronological Aging in Saccharomyces cerevisiae. Anal Chem 90: 13506-13513 (2018).
2019 Castells-Nobau A, Eidhof I, Fenckova M, Brenman-Suttner DB, Scheffer-de Gooyert JM, Christine S, Schellevis RL, van der Laan K, Quentin C,
van Ninhuijs L, Hofmann F, Ejsmont R, Fisher SE, Kramer JM, Sigrist SJ, Simon AF, Schenck A. Conserved regulation of neurodevelopmental
218 219
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Processed on: 21-1-2020 PDF page: 220PDF page: 220PDF page: 220PDF page: 220 processes and behavior by FoxP in Drosophila. PLoS One 14: e0211652
(2019).
Morita A, Perona Martinez FP, Chipaux M, Jamot N, Hemelaar SR, van der Laan KJ, Schirhagl R. Cell Uptake of Lipid-Coated Diamond. Part
Part Syst Char 36: 1900116 (2019).
Sigaeva A, Ong Y, Damle VG, Morita A, van der Laan KJ, Schirhagl R.
Optical Detection of Intracellular Quantities Using Nanoscale Technologies. Acc Chem Res 52: 1739-1749 (2019).
