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  1. 9 Challenges of Modern Technology
  2. 5 Polish Journal of Chemistry
  3. 4 Elektronika : konstrukcje, technologie, zastosowania
  4. 2 Analityka : nauka i praktyka
  5. 2 Biocybernetics and Biomedical Engineering
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  1. 32 Brzózka Z.
  2. 18 Chudy M.
  3. 18 Dybko A.
  4. 6 Wróblewski W.
  5. 4 Jędrych E.
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  1. 1 2015
  2. 2 2013
  3. 3 2012
  4. 4 2011
  5. 3 2010
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1
Content available Inżynieria komórkowa w systemach lab-on-a-chip
Tomecka E., Tokarska K., Jastrzębska E., Chudy M., Brzózka Z.
Wiadomości Chemiczne
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2015
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[Z] 69, 9-10
909--929
EN
Lab-on-a-chip systems are promising tools in the field of cell engineering. Microfluidic systems are integrated microlaboratories consisting of many microstructures such as microchannels and microchambers, which can be used for cell analysis and cell culture. Appropriately designed geometry of the chip allows to mimic in vivo conditions. Microsystems enables continuous culture medium perfusion. During cell culture, regulation of the flow rate of medium is possible, which allows to control conditions of the cultivation. In this paper we present a review of microfluidics systems which are used in cell engineering. We describe methods of microsystems fabrication, parameters which influence cell proliferation in microscale and examples of microsystems for cell analysis and cell culturing. Microfluidic systems for maintaining cell culture are mainly fabricated of poly(dimethylsiloxane) (PDMS) and glass, non-toxic materials for cells. The most commonly used method for fabrication of PDMS microsystems is photolithography and replica molding techniques. Cell culture in microsystems can be carried out in two ways: as a two-dimensional (2D) cell culture and three-dimensional (3D) cell culture. In two-dimensional culture cells grow as a monolayer on a flat surface of microchambers or microchannels. Microsystems for two-dimensional cell culture are widely described in the literature. They are mainly used for: (i) cell proliferation after exposure to external stimuli, (ii) testing the activity of cytotoxic drugs, (iii) interactions and cell migration and (iv) the evaluation of procedures applicable in tumor therapy e. g. photodynamic therapy. However, two-dimensional cell culture do not mimic fully in vivo conditions. In living organisms cells grow spatially creating three-dimensional structures like tissues. Therefore, nowadays microsystems for 3D cell culture are being developed intensively. Three-dimensional cell culture in microfluidic systems can be achieved in three ways: by the design of suitable geometry and topography of microchannels, by the use of hydrogels or by spheroids formation. Three-dimensional cell culture in microfluidic systems are much better experimental in vitro models than cell culture in traditional culture vessels. It is the main reason why microsystems should be still improved, as to become widely used research tools in cellular engineering.
2
Content available Potentiometric detection of the metabolic activity of human tumor cells
Kwapiszewska K., Stępień K., Sendys P., Chudy M., Brzózka Z.
Challenges of Modern Technology
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2013
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Vol. 4, no. 1
41--46
EN
Monitoring of cellular viability is a key part of toxicological assays in vitro. On-line monitoring of metabolic activity would be particularly useful for evaluation of responses to potential therapeutic compounds. Current assays are mostly based on fluorescent dyes and optical detection methods. These methods offer high sensitivity and specificity, however are not suitable for long-term on-line observations. Electrochemical methods can be an alternative for current protocols. Electrochemical detection is low cost and label-free, therefore suitable for long-term cell culture monitoring. In this work investigations on human cancer cells viability will be presented. Cells were cultured as two-dimensional monolayer or three-dimensional spheroids. Different cell culture media were examined. Potentiometric detection was used for continuous monitoring of cell culture as well as end-point investigations. Different growth phases were identified using applied method. Finally, response to an anticancer drug was successfully observed.
3
Content available An integrated gas removing system for microfluidic application. Design and evaluation
Hofman I. M., Ziółkowska K., Dybko A., Brzózka Z.
Challenges of Modern Technology
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2013
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Vol. 4, no. 3
10--15
EN
Microfluidic systems are used in a wide range of applications, including medical diagnostics, cell engineering and bioanalytics. In this work we focused on “Lab-on-a-chip” microsystems for cell cultivation. A troublesome problem of gas bubbles entering microdevices causing signal interferences and cells damage was emphasized. A novel, integrated debubbler in the form of cylindrical traps covered with thin PDMS membrane was designed and manufactured. Demonstrated debubbler was successfully applied in a long-lasting culture of HT-29 cell aggregates.
4
Content available New microfluidic device for lactate dehydrogenase (LDH) activity analysis
Jędrych E., Mazur M., Grabowska-Jadach I., Brzózka Z.
Challenges of Modern Technology
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2012
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Vol. 3, no. 4
3--8
EN
In this paper, we present cytotoxicity analysis (determination of lactate dehydrogenase — LDH activity performed in a designed and fabricated microfl uidic system. This method allowed for analysis of a supernatant collected from A549 (human lung cancer) and HT-29 (human colon cancer epithelial) cells, which were incubated for 24 h with selected compounds. LDH is an intracellular enzyme present in tissues, which is released into the supernatant caused by membrane damage or cell lyses. In our tests, LDH-Cytotoxicity Assay Kit (BioVision) was used. The toxic eff ect of drugs was measured in the developed microsystem made of PDMS (poly(dimethylsiloxane)). Analytical reaction took place in the special designed microchannel geometry. Then, the LDH activity was measured at 490 nm using spectrophotometer. In subsequent experiments, appropriate conditions for measurements using a microfl uidic system were chosen. It was found that the selected reagent is sensitive to temperature changes and light exposure. Reaction time in the microsystem was determined by changes of fl ow rates of reagents. Afterwards, for the various reaction time, the toxic eff ect of 5-fl uorouracil, celecoxib and 1,4-dioxane was performed. The obtained results were compared with the results carried out in 96-well plates. Based on these results, it was noted that the enzymatic reaction time in the microsystem is shorter than in 96-well plate. Moreover, the advantage of using microsystem is also the small amount of reagents.
5
Content available Microfluidic devices — application in anticancer studies
Jędrych E., Chudy M., Dybko A., Brzózka Z.
Challenges of Modern Technology
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2012
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Vol. 3, no. 2
3--5
EN
A rapidly growing pharmaceutical industry requires faster and more efficient ways to find and test new drugs. One of the new method for cell culture and examining the toxic effects of drugs is application of microfluidic systems. They provide new types of microenvironments and new methods for investigation of anticancer therapy. The use of microsystems is a solution that gives the opportunity to reduce not only cost and time, but also a number of tests on animals. In this paper we present designed and fabricated hybrid microfluidic systems which are applicable for cell culture, cell based cytotoxicity assays and photodynamic therapy procedures. Polydimethylsiloxane (PDMS) and sodium glass were used for fabrication of microdevices. The designed geometry of the microdevices includes cell culture microchambers and a concentration gradient generator (CGG). The CGG enables to obtain diff erent concentrations of tested drugs in a single step, which is a significant simplification of cytotoxicity assay procedure. In the designed microsystems three various cell lines (normal and carcinoma) were cultured and analyzed.
6
Content available Development of a Point-of-Care system for early diagnostics of genetic diseases
Kwapiszewski R., Chudy M., Dybko A., Brzózka Z.
Challenges of Modern Technology
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2012
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Vol. 3, no. 2
6--8
EN
Lysosomal storage disorders (LSDs) represent a group of more than 45 genetically inherited diseases caused by the absence or deficiency of one or more specific lysosomal enzymes. Nowadays, there is a lack of reports on fast, reliable methods for the diagnostics of LSDs. Currently applied diagnostic approaches generate many false-negative and false-positive results, which results in classification of patients to inappropriate therapeutic groups. Moreover, these methods are time-consuming (even 20 hours), and are carried out only in a few laboratories in the world. The goal of this work was to develop a method and a tool, a Point-of-Care system, for diagnostics of LSDs. The polymeric microdevice consists of a cell lysis module, a mixing microchannel and an optical detection module. The system enables to determine the activity of α-galactosidase (deficient in Fabry disease), and to reduce the time of analysis to 10 min. Due to its easy fabrication steps and low price, it seems to be a prospective tool for a point-of-care approach.
7
Content available Novel designs and technologies for cell engineering
Ziółkowska K., Chudy M., Dybko A., Brzózka Z.
Challenges of Modern Technology
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2011
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Vol. 2, no. 4
54--60
EN
Microfluidic devices, such as lab-on-a-chip systems, are highly advantageous for cell engineering and cell based assays. It is a particularly useful approach for development of the in vitro cellular systems mimicking the in vivo environment. In this paper, a novel lab-on-a-chip device for three-dimensional human cell culture and anticancer drug testing is presented. Cells were cultured as Multicellular Tumor Spheroids (MCTS) — the best cancer tumor model developed so far. Diff erent designs were tested and novel technique of microfabrication in poly(dimethylsiloxane) was developed. MCTS were cultured in a system of polymeric microwells, with the network of microfluidic channels for culture medium flow. Design included optimal shear stress and proper nutrients supply for cultured cells. Final design provided MCTS culture for four weeks with the homeostasis-like state achievement, which is characteristic for the in vivo situation.
8
Content available Miniaturized device for a cell lysis process
Kwapiszewski R., Chudy M., Dybko A., Brzózka Z.
Challenges of Modern Technology
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2011
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Vol. 2, no. 2
50--52
EN
Single-cell studies are crucial for gaining knowledge on complexity of intracellular processes. In many cases, carrying researches into cell ingredients must be proceeded by a lysis process. Cell lysis leads to disintegration of the plasma membrane which is the barrier separating cell contents from the environment. However, investigations at the cellular level would not be possible without proper miniaturized tools, which offer many advantages as low reagents consumption, short reaction time, integration, automation or versatility. The goal of this work was to design and develop a microfluidic chip for a chemical cell lysis process. The geometry of a microsystem presented is based on the hydrodynamic focusing of a cell suspension stream. Applying non-denaturing cell lysis buffer enables to analyze released cell ingredients during next steps of investigations.
9
Content available ‘Lab-on-a-chip’ for cell engineering: towards cellular models mimicking in vivo
Ziółkowska K., Chudy M., Dybko A., Brzózka Z.
Challenges of Modern Technology
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2011
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Vol. 2, no. 1
79--82
EN
One of the main scopes of modern cell engineering is development of cellular models that can replace animals in drug screening and toxicological tests, so called alternative methods. Construction of the alternative model is a very challenging task due to a richness of factors creating the in vivo environment. The monolayer cell culture — cultivation of adhesive cells on artificial surfaces such as glass or polymer — lack most of the in vivo-like interactions, but still is the only tool for the majority of applications. One of the most prospective approaches on mimicking in vivo environment is “Lab-on-a-chip” technology. Microfluidic devices offer lots of advantages over traditional in vitro culture, e.g. much higher cell volume-to-extracellular fluid volume ratio or possibility of regulation of hydrodynamic stress. This presentation aims to introduce latest advances of our team in microfluidic cell culture devices. Our novel approach is to cultivate three dimensional multicellular aggregates (spheroids) in microenvironments arranged in a microfluidic system. The geometry and materials of the system allow for cultivation, observation and analysis of multicellular spheroids. The results presented concern multicellular tumor spheroids (MCTS) rising from human cancer cells, which are considered to represent most of the conditionings of cancer tumor in vivo. The fully developed MCTS microdevice will be a reliable tool for anticancer drug screening, as the results most likely will be in a close accordance with the results obtained in vivo.
10
Profesor Jan Czochralski - historia najwybitniejszego polskiego inżyniera
Rachoń J., Brzózka Z.
Analityka : nauka i praktyka
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2011
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nr 3
56-57
11
Content available remote Pasywny mikromieszalnik przepływowy wytworzony metodą mikrofrezowania i bondowania termicznego w PMMA
Żukowski K., Chudy M., Dybko A., Brzózka Z.
Przegląd Elektrotechniczny
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2010
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R. 86, nr 10
154-156
PL
Celem prezentowanej pracy było wykonanie pasywnego, wysokosprawnego mikromieszalnika przepływowego. Wykonany mikromieszalnik był strukturą w pełni trójwymiarową, w której mieszanie zachodzi głównie poprzez dyfuzję. Mikrokanały wykonano metodą mikrofrezowania w płytkach z PMMA, a następnie połączono ze sobą poprzez bondowanie termiczne. Parametry procesu bondowania dobrano w ten sposób, aby zminimalizować deformację mikrokanałów. Dodatkowo opracowano szybką metodę justowania mikrokanałów.
EN
This paper presents a highly efficient passive micromixer with a three-dimensional topography of microchannel that employs diffusion for mixing. The microchannels were created in PMMA plates by mechanical milling with a high frequency spindle. The PMMA plates with microchannels were thermally bonded to form sealed 3D micromixer structure. The developed device is easy to fabricate and has excellent working characteristics in the continuous-flow mode. In this paper we describe a simple method of microchannels’ adjustment.
12
Content available remote Miniaturowy system analityczny do hodowli komórkowych
Jędrych E., Ziółkowska K., Chudy M., Brzózka Z.
Przegląd Elektrotechniczny
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2010
|
R. 86, nr 10
33-35
PL
Celem badań było wykonanie miniaturowego systemu przeznaczonego do hodowli komórek adherentnych, a następnie ocena jego przydatności w prowadzeniu analiz z wykorzystaniem hodowanych komórek. W wykonanym mikrosystemie przeprowadzono kilkudniową hodowlę komórek: A549 – komórki nowotworowe płuc oraz HT-29 – komórki nowotworowe jelita grubego, wcześniej opracowując protokoły sterylizacji i przygotowania mikroukładu do hodowli. Przeprowadzono testy związane z oceną toksyczności komórek na modelowym związku, jakim był 1,4- dioksan. Prawidłowość pracy mikroukładu potwierdzono poprzez skorelowanie otrzymanych wyników z testem wykonanym w skali makroskopowej. Opracowano również protokół ponownego wykorzystania mikrochipu do hodowli komórkowej oraz metodykę oceny absorpcji w PDMS-ie testowanych związków (dioksanu) w mikroukładzie.
EN
The aim of our study was to obtain a microsystem for adherent cell culture and then assess its usefulness for tests on the cultured cells. The culture of the human lung carcinoma cells (A549) and colorectal carcinoma cells (HT-29) was carried on in the microdevice for several days. The microdevice before cell docking in microchambers was especially prepared and sterilized according to the developed procedures. Both A549 and HT-29 cells are adherent, however they require a different conditions of growth to be optimized. However, the possibility of using the chip constructed and designed for culture different cell lines has been confirmed, which underline the versatility of the microchip. The suitability of the designed microsystem for cell-based cytotoxicity assay application was verified using 1,4-dioxane as a model toxic agent. The series of cytotoxicity tests in the microdevice as well as in classic 96-well cell culture plates were performed to compare results obtained in micro- and macroscale. The microdevice is fully reusable, i.e. it was used several times for various cell culture and cytotoxic experiments. The methodology for assessing of the absorption of tested compounds (1,4-dioxane) in PDMS in the chip was developed.
13
Content available Photodynamic therapy procedures in the microfluidic system
Jędrych E., Pawlicka Z., Chudy M., Dybko A., Brzózka Z.
Challenges of Modern Technology
|
2010
|
Vol. 1, no. 1
30--33
EN
Evaluation of the effi ciency of photodynamic therapy (PDT) in a hybrid microfl uidic culture system was studied. The geometry of the utilized microsystem for PDT procedures consists microchambers for cell culture and microchannels, which create a concentration gradient generator (CGG). 5-aminolevulinic acid (ALA) as a precursor of the photosensitizer was used. The geometry of the microchip allowed to test diff erent concentrations of ALA in a single assay. Evaluation of the effi ciency of photodynamic therapy was determined 24 hours after PDT procedure (irradiation with light which induced accumulated in carcinoma cells). The performed microsystem contained two independent micropatterns, that enables examination simultaneously various cell lines (carcinoma and normal) or various photosensitizers.
14
Szybka metoda wykonywania niebondowanych mikroukładów w technologii polimerowej
Juchniewicz M., Chudy M., Brzózka Z., Dybko A.
Elektronika : konstrukcje, technologie, zastosowania
|
2008
|
Vol. 49, nr 6
75-76
PL
Zaprezentowano wyniki badań nad opracowaniem nowej technologii wykonywania niebondowanych przepływowych mikroukładów chemicznych. Z filmu kapilarnego w procesie fotolitografii wykonano matryce. Zostały one użyte w procesie odlewania do wykonania nowego typu niebondowanego mikroukładu. Niebondowane mikroukłady wykonano przez zatopienie w strukturze polimeru matrycy z emulsji filmu kapilarnego. Po usieciowaniu polimeru emulsję usunięto z jego struktury pozostawiając sieć mikrokanatów.
EN
The paper presents results of developing of a new technology for fabrication of bonding-less (B-Less) microfluidic chemical systems. The matrixes for the microsystem preparation were made of a capilla film. They were used in a moulding process for a fabrication of B-Le: microsystem. For the fabrication of B-Less microsystem an emulsk of a capillary film was moulded inside of a polymer block. After p lymer crosslinking the emulsion was removed from the block and the microchannels were formed.
15
Nowy typ przepływowego mikrokonduktometru bezkontaktowego
Juchniewicz M., Ekielski M., Chudy M., Brzózka Z., Dybko A.
Elektronika : konstrukcje, technologie, zastosowania
|
2008
|
Vol. 49, nr 6
87-88
PL
Przedstawione wyniki dotyczą prac nad konstrukcją mikrokonduktometru przepływowego z nowym typem elektrod. Opisano wykonanie mikrokonduktometrów z dwóch materiałów: szkła i polimeru oraz z połączenia tych materiałów. W konstrukcji wykorzystano i porównano ze sobą trzy rodzaje elektrod pomiarowych: srebrzone mikrokanały, mikrokanały wypełnione elektrolitem i mikrokanały jednocześnie posrebrzone i wypełnione elektrolitem. Działanie wykonanych mikrokonduktometrów sprawdzono podczas pomiarów przewodnictwa wzorcowych roztworów KCI o różnym stężeniu.
EN
Presented results apply for works on construction of microfluidic conductometer with a new type of electrodes. Fabrication of conductometer from two materials was described: glass, polymer and combination of both of these materials. For construction, three types of electrodes were applied and compared: silver plated microchannels, microchannels filled with an electrolyte and microchannels simultaneously silver plated and filled with an electrolyte. Fabricated microconductometers were tested during the measurements of standard solutions with a different KCI concentrations.
16
Technika łączenia warstw polimerowych z ceramicznymi elementami mikroukładów
Chudy M., Malecha K., Golonka L., Sosicki A., Roguszczak H., Jakubowska M., Dybko A., Brzózka Z.
Elektronika : konstrukcje, technologie, zastosowania
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2007
|
Vol. 48, nr 3
13-14
PL
Przedstawiono możliwości konstrukcji mikroukładów chemicznych z wykorzystaniem materiałów ceramicznych i polimerowych. Opracowano technikę trwałego i odwracalnego łączenia warstw ceramicznych i polimerowych, zależnie od dalszego przeznaczenia konstruowanego mikroukładu. W przypadku wykorzystania go w systemie z ciśnieniowym transportem mediów ciekłych przez układ konieczne jest bondowanie trwałe z wykorzystaniem plazmowej modyfikacji kolejnych warstw. W mikroukładach z transportem elektroosmotycznym próbek wystarczają jedynie siły wzajemnej adhezji warstw ceramicznych i polimerowych. W obu przypadkach ceramiczne elementy mikrosystemów należało pokryć warstwą szkliwa.
EN
The possibilities of the construction of microsystems using ceramics and polymers were presented in the paper. The technology of irreversible and reversible bonding of ceramic and polymer microsystems' layers was developed. The irreversible bonding is required only for microfluidic structures, in which samples and reagents are introduced into the system using pressure methods. For the systems with an electroosmotic reagents dosing adhesion forces between particular layers are enough to seal the microchannels. In both cases a glaze layer was screen-printed on ceramic plates to eliminate their surface roughness.
17
Microcapillary Electrophoresis with Fluorescence Detection
Lewandowska N., Dybko A., Chudy M., Wcisło M., Poboży E., Brzózka Z.
Polish Journal of Chemistry
|
2006
|
Vol. 80, nr 11
1799-1806
EN
The advantages and application perspectives for microcapillary electrophoresis (mi CE) are presented. The microchip based on a glass plate was fabricated. The separations of three FITC-labeled amino acids carried out with the conventional CE system and in the microchip were compared. The obtained results proved the usability of the microchip for mi CE process.
18
Fabrication of ceramic-polymer microstructure for capillary electrophoresis with usage of photosensitive paste and thick film technology
Wyżkiewicz I., Lewandowska N., Dziurdzia B., Magoński Z., Chudy M., Brzózka Z., Jakubowska M., Dybko A.
Elektronika : konstrukcje, technologie, zastosowania
|
2006
|
Vol. 47, nr 12
9-10
EN
Fabrication of a ceramic-polymer hybrid structure with microchannel was accomplished. The structure for capillary electrophoresis is presented as an example system. The method of fabrication of such a structure is not complicated, makes use of well known technologies like thick film and photolithography, and commercially available materials. It could be used for manufacture of structures with different shapes and dimensions. The process relies on printing dielectric photosensitive paste onto a ceramic support, drying and exposing to UV light through a mask with suitable pattern, developing and firing. A thin glaze layer is deposited on the top of the structure in order to make possible reversible or irreversible bonding with transparent poly(dimethylsiloxane) (PDMS) and adapting this chip to measurements with the use of optical detection.
PL
Opracowano wytwarzanie ceramiczno-polimerowych struktur zawierających mikrokanał. Jako przykład zaprezentowano strukturę do elektroforezy kapilarnej. Metoda wytwarzania takich mikroukładów jest metodą prostą, wykorzystującą dobrze znane technologie, takie jak technologia grubowarstwowa czy fotolitografia i materiały komercyjnie dostępne. Może być wykorzystywana do wytwarzania struktur hybrydowych o różnej geometrii kanałów. Proces wytwarzania mikroukładów polega na nadrukowaniu światłoczułej pasty dielektrycznej na podłoże ceramiczne, wysuszeniu i naświetleniu przez fotomaskę z odpowiednim wzorem, wymyciu nienaświetlonej części warstwy i wypaleniu. Pokrycie struktury cienką warstwą szkliwiącą daje możliwość odwracalnego lub trwałego łączenia z transparentnym poli(dimetylosiloksanem) (POMS) i dostosowania takiego układu do pomiarów z detekcją optyczną.
19
Content available remote Spectrophotometric analysis using poly( dimethylsiloxane) microfl uidic detectors
Stadnik D., Chudy M., Brzózka Z., Dybko A.
Bulletin of the Polish Academy of Sciences. Technical Sciences
|
2005
|
Vol. 53, nr 2
163-165
EN
Two constructions of microfluidic structures are described in this paper. A fibre optic micro cell for spectrophometric measurements and a micro cell for fluorescence experiments were designed and tested. The structures were made of polymer optical fibres which were incorporated into polymeric material Le. poly( dimethylsiloxane). The structures were tested as detectors in absorbance measurement (solutions of bromothymol blue with different pH were used) and in fluorescence tests (solution of fluoresceine was used).
20
Design of Ferrocene Organothiol Monolayer as Intermediate Phase for Miniaturized Electrochemical sensors with Gold Contact
Sęk S., Bilewicz R., Grygołowicz-Pawlak E., Grudzień I., Brzózka Z., Malinowska E.
Polish Journal of Chemistry
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2004
|
Vol. 78 / nr 9
1655-1665
EN
New design of back side contact (BSC) chips for miniaturized electrochemical sensors with Au contact modified by monolayers of purposely synthesized organothiol molecules is described. Voltammetric studies of the monolayers indicate reversible behavior of the compounds and efficient coverage of the gold electrode. Properties of 5 compounds are compared. The electroactive benzenethiol based self-assembled monolayer is selected as a convenient intermediate layer in solid contact potentiometric sensors since it forms a hydrophobic barrier between the electrode and the solution and at the same time provides efficient electronic wiring to the electrode. Stable electrode performance is obtained and the potentiometric response remains as fast as using bare gold contact which should be favorable for the application of the described intermediate layers in ion-selective electrodes.
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