Experimental facility: Difference between revisions

Latest revision as of 18:14, 11 December 2023

Our experimental facility consists of 11 rooms with the total area of ca. 200 m². Each room is equipped with air conditioning, air humidity control system, and connected to an automatic stand-by power generator. Additionally, Tissue Culture Room, Molecular Biology Unit and Microfluidics Unit are equipped with HEPA H14 air filtration system.

We have the permission for the contained use of GMOs in our research.

Tissue Culture Unit

Tissue culture room.

laminar hoods
CO₂ incubators
inverted laboratory microscope (Leica DM IL LED)
automated cell counter (Bio-Rad TC10 Automated Cell Counter)
water baths
shaker
incubated shaker
centrifuge
micro-centrifuge
freezer −25 °C
ultra-low temperature freezer −86 °C
cryogenic storage Dewar
autoclave 108L (HMC HG113)
water purification system
icemaker
accessories: vortex, magnetic stirrer, heating block, aspirators

Molecular Biology Unit

Molecular biology room.

laminar hood
CO₂ incubator
inverted laboratory microscope (Leica DM IL LED)
bioanalyzer (Agilent 2100 Bioanalyzer)
nucleoporator (Lonza Nucleofector IIS)
PCR thermocycler (Eppendorf Mastercycler)
real-time PCR (Applied Biosystems QuantStudio 12K Flex)
digital PCR (Applied Biosystems QuantStudio 3D)
flow cytometer (BD FACSCalibur)
fluorometer (Thermo Scientific Fluoroskan Ascent Microplate Fluorometer)
spectrophotometer (Thermo Scientific Multiskan GO Microplate Spectrophotometer)
Western blot set
nucleic acid gel separation system
imaging system (Bio-Rad ChemiDoc MP Imaging System)
gel imaging system (Bio-Rad Molecular Imager GelDoc XR+)
centrifuge
micro-centrifuge
water baths
refrigerator
accessories: vortex, magnetic stirrer, balance, heating block
dark-room (separate)

Microscopy Unit

Confocal microscopy unit.

confocal microscope Leica TCS SP5 X with environmental chamber
- based on inverted, fully automated microscope Leica DMI6000
- environmental chamber with temperature, CO₂ and humidity control
- scanning stage with Super-Z Galvo
- 3 PMT confocal detectors
- 1 brightfield detector
- digital camera Leica DFC365FX (cooled, 21 fps at full resolution 1392×1040)
- fluorescence light source: 120 W mercury metal halide lamp
- Lasers:
  - White Light Laser (WLL): tunable excitation wavelengths from 470 nm to 670 nm
    with 1 nm resolution, up to 8 excitation lines can be used simultaneously
  - diode 405 nm
  - multi-wavelengths Argon laser: 458, 476, 488, 496 and 514 nm
  - DPSS 561 nm
  - HeNe 633 nm
- Objectives:
  - HC PL FLUOTAR 5×/0.15
  - HC PL APO 10×/0.40 CS
  - HC PL APO 20×/0.70 CS
  - HCX PL APO 40×/0.85 CS
  - HCX PL APO 63×/1.40-0.60 OIL CS
- Fluorescence filter cubes:
  - GFP (filter "GFP"): excitation: BP470/40, dichroic: 495, emission: BP525/50
  - FITC (filter "I.3"): excitation BP450-490, dichroic: 510, emission: LP515
  - TRITC (filter "N2.1"): excitation BP515-560, dichroic: 580, emission: LP590
  - Red "band-pass" (filter "N3 ET"): excitation BP546/11, dichroic: 565,
    emission:BP 600/40
  - DAPI "long-pass" (filter "A"): excitation BP340-380, dichroic: 400, emission: LP425
  - DAPI "band-pass" (filter "A4 ET"): excitation BP360/40, dichroic: 400,
    emission: BP470/40

TIRF microscope Leica AM TIRF MC
- based on inverted, fully automated microscope Leica DMI6000 with AFC
- environmental chamber with temperature, CO₂ and humidity control
- methods: TIRF, FRET, fluorescence, bright filed, DIC, polarization
- fully integrated module for Multi-Color Total Internal Reflection Fluorescence
  - specimen penetration depth from 70 nm to 300 nm
  - lasers: 405, 488, 561 and 635 nm with AOTF
  - TIRF objective: HCX PL APO 100×/1.47 OIL CORR TIRF
- fluorescence light source: 120 W mercury metal halide lamp
- Cameras:
  - EM-CCD, Hamamatsu C9100-02 (1000×1000 px, monochrome, cooled, high EM gain, up to 256 fps, 31 fps at full resolution)
  - Leica DFC365FX (1391×1040 px, monochrome, cooled, up to 122 fps, 21 fps at full resolution)
  - Leica DFC295 (2048×1536 px, color, up to 35 fps, 12 fps at full resolution)
- Objectives:
  - N PLAN 2.5×/0.07
  - N PLAN 5×/0.12
  - HCX PL S-APO 10×/0.30
  - HCX PL FL L 20×/0.40 CORR
  - HCX PL FL L 63×/0.70 CORR
  - HCX PL APO 100×/1.47 OIL CORR TIRF

high-throughput microplate imager: Operetta CLS (PerkinElmer)
confocal bio-imaging system: BD Pathway 435 (BD Biosciences)
integrated system of atomic force microscopy (AFM), confocal microscopy and Raman spectroscopy (NTEGRA Spectra, NT-MDT Co.)
inverted fluorescent microscope (DMI3000B, Leica)
optical tweezers

Microfluidics Unit

Microfluidics unit.

microfluidic platform for cell culturing CellASIC ONIX
microPIV platform: Nikon Eclipse E-50i epi-fluorescent microscope with double-shutter CCD camera and double-pulsed Nd:YAG laser
stereo microscopes
high-precision syringe pumps (neMESYS, Cetoni GmbH)
high-precision peristaltic pumps (peRISYS, Cetoni GmbH)
syringe pumps, peristaltic pump, micro gear pump
high-speed intensified CMOS camera (Lambert HiCAM 500)
high-speed camera (Photron, FASTCAM Mini AX100 540K-M-8GB)
high-speed CMOS camera (pco.1200 hs, PCO IMAGING)
double-shutter CCD camera (SensiCam, PCO IMAGING)
equipment for microfluidic chip fabrication by soft photolithography, i.a. spin coater, plasma cleaner, UV curing system, laboratory ovens, heating plates
equipment for microfluidic chip fabrication by micro-milling
3D printer (Zortrax Inventure)

Biomaterials Unit

setup for polymer nanofibers fabrication by electrospinning equipped with environmental chamber
3D bioprinter (Cellink, INKREDIBLE S-10002-001)
lyophilizator (LABCONCO, FreeZone 2.5 Liter -84C Benchtop Freeze Dryer 710201030)
plasma cleaner (Diener, ZEPTO PCCE)
fume hood
accessories: analytical balances, centrifuges, vortexes, magnetic stirrers, blood roller mixer, water baths, refrigerated/heating circulators, conductometer, pH-meter, vacuum pump, pipettes, etc.

Other equipment

laser plotter equipped with Jasper X0-30 femtosecond laser (Fluence, Poland), designed for micromachining, cutting, and surface structuring of a wide range of materials (e.g., glass, plastics)
high-energy, double-pulsed Nd:YAG nanosecond laser (EKSPLA NL303D)
high frequency 1 kHz pulsed Nd:YAG laser (EKSPLA NL202)
double-pulsed Nd:YAG laser (SoloPIV, New Wave Research Inc.)
solenoid micro-valves
pressure sensors
temperature sensors
accessories like camera lenses, microscope objectives, mirrors, lenses, halogen lamps, sensors of temperature, pressure, humidity, viscosity and others

Laboratory equipment was partially financed by the European Union – the European Regional Development Fund within the Operational Programme “Innovative Economy” for 2007–2013.

@@ Line 1: / Line 1: @@
-Our experimental facility consists of 9&nbsp;rooms with the total area of ca. 160&nbsp;m<sup>2</sup>. Each room is equipped with air conditioning, air humidity control system, and connected to an automatic stand-by power generator<!--running water and high-power electrical connections-->. Additionally, Tissue Culture Room, Molecular Biology Unit and Microfluidics Unit are equipped with ''HEPA H14'' air filtration system.
+Our experimental facility consists of 11&nbsp;rooms with the total area of ca. 200&nbsp;m<sup>2</sup>. Each room is equipped with air conditioning, air humidity control system, and connected to an automatic stand-by power generator<!--running water and high-power electrical connections-->. Additionally, Tissue Culture Room, Molecular Biology Unit and Microfluidics Unit are equipped with ''HEPA H14'' air filtration system.
-We [{{SERVER}}/documents/pozwolenie_na_zamkniete_uzycie_GMO.pdf have permission] for the [http://ec.europa.eu/food/plant/gmo/authorisation/contained/index_en.htm contained use] of GMOs in our research.
+We have [{{SERVER}}/documents/pozwolenie_na_zamkniete_uzycie_GMO.pdf the permission] for the [https://food.ec.europa.eu/plants/genetically-modified-organisms/gmo-authorisation/contained-use_en contained use] of GMOs in our research.
+===Tissue Culture Unit===
-===Tissue Culture Room===
 [[Image:Experimental_facility_cell_culture_room.jpg|link={{SERVER}}/images/b/bf/Experimental_facility_cell_culture_room.jpg||thumb|Tissue culture room.]]
 *laminar hoods
@@ Line 30: / Line 29: @@
 *CO<sub>2</sub> incubator
 *inverted laboratory microscope (''Leica DM IL LED'')
+*bioanalyzer (''Agilent 2100 Bioanalyzer'')
 *nucleoporator (''Lonza Nucleofector IIS'')
 *PCR thermocycler (''Eppendorf Mastercycler'')
@@ Line 39: / Line 39: @@
 *Western blot set
 *nucleic acid gel separation system
+*imaging system (''Bio-Rad ChemiDoc MP Imaging System'')
 *gel imaging system (''Bio-Rad Molecular Imager GelDoc XR+'')
 *centrifuge
@@ Line 50: / Line 51: @@
 ===Microscopy Unit===
 [[Image:Experimental_facility_microscopy_unit.jpg|link={{SERVER}}/images/3/37/Experimental_facility_microscopy_unit.jpg|thumb|Confocal microscopy unit.]]
-* confocal  microscope ''Leica&nbsp;TCS SP5&nbsp;X'' with environmental chamber&nbsp;<div class="toccolours mw-collapsible mw-collapsed" style="width:290px"><div class="mw-collapsible-content">
+* confocal  microscope ''Leica&nbsp;TCS SP5&nbsp;X'' with environmental chamber&nbsp;<div class="toccolours mw-collapsible mw-collapsed"><div class="mw-collapsible-content">
 ** based on inverted, fully automated microscope Leica DMI6000
 ** environmental chamber with temperature, CO<sub>2</sub> and humidity control
@@ Line 65: / Line 66: @@
 *** HeNe 633&nbsp;nm
 ** '''Objectives''':
-*** HC PL FLUOTAR 5x/0.15
+*** HC PL FLUOTAR 5&times;/0.15
-*** HC PL APO 10x/0.40 CS
+*** HC PL APO 10&times;/0.40 CS
-*** HC PL APO 20x/0.70 CS
+*** HC PL APO 20&times;/0.70 CS
-*** HCX PL APO 40x/0.85 CS
+*** HCX PL APO 40&times;/0.85 CS
-*** HCX PL APO 63x/1.40-0.60 OIL CS
+*** HCX PL APO 63&times;/1.40-0.60 OIL CS
 ** '''Fluorescence filter cubes''':
 *** GFP (filter "GFP"): excitation: BP470/40, dichroic: 495, emission: BP525/50
@@ Line 78: / Line 79: @@
 *** DAPI "band-pass" (filter "A4 ET"): excitation BP360/40, dichroic: 400, <br>emission: BP470/40</div></div>
-* TIRF microscope ''Leica&nbsp;AM&nbsp;TIRF&nbsp;MC''&nbsp;<div class="toccolours mw-collapsible mw-collapsed" style="width:290px"><div class="mw-collapsible-content">
+* TIRF microscope ''Leica&nbsp;AM&nbsp;TIRF&nbsp;MC''&nbsp;<div class="toccolours mw-collapsible mw-collapsed"><div class="mw-collapsible-content">
 ** based on inverted, fully automated microscope Leica&nbsp;DMI6000 with AFC
 ** environmental chamber with temperature, CO<sub>2</sub> and humidity control
 ** methods: TIRF, FRET, fluorescence, bright filed, DIC, polarization
-** fully integrated module for Multi-Color Total Internal Reflection Fluorescence technique
+** fully integrated module for Multi-Color Total Internal Reflection Fluorescence
 *** specimen penetration depth from 70&nbsp;nm to 300&nbsp;nm
 *** lasers: 405, 488, 561 and 635&nbsp;nm with AOTF
-*** TIRF objectiv: HCX PL APO 100x/1.47 OIL CORR&nbsp;TIRF
+*** TIRF objective: HCX PL APO 100&times;/1.47 OIL CORR&nbsp;TIRF
 ** fluorescence light source: 120&nbsp;W mercury metal halide lamp
 ** '''Cameras''':
-*** EM-CCD, Hamamatsu C9100-02 (1000x1000&nbsp;pix, monochrome, cooled, high EM&nbsp;gain, up to 256&nbsp;fps, 31&nbsp;fps at full resolution)
+*** EM-CCD, Hamamatsu C9100-02 (1000&times;1000&nbsp;px, monochrome, cooled, high EM&nbsp;gain, up to 256&nbsp;fps, 31&nbsp;fps at full resolution)
-*** Leica DFC365FX (1391x1040&nbsp;pix, monochrome, cooled, up to 122&nbsp;fps, 21&nbsp;fps at full resolution)
+*** Leica DFC365FX (1391&times;1040&nbsp;px, monochrome, cooled, up to 122&nbsp;fps, 21&nbsp;fps at full resolution)
-*** Leica DFC295 (2048x1536&nbsp;pix, color, up to 35&nbsp;fps, 12&nbsp;fps at full resolution)
+*** Leica DFC295 (2048&times;1536&nbsp;px, color, up to 35&nbsp;fps, 12&nbsp;fps at full resolution)
 ** '''Objectives''':
-*** N PLAN 2.5x/0.07
+*** N PLAN 2.5&times;/0.07
-*** N PLAN 5x/0.12
+*** N PLAN 5&times;/0.12
-*** HCX PL S-APO 10x/0.30
+*** HCX PL S-APO 10&times;/0.30
-*** HCX PL FL L 20x/0.40 CORR
+*** HCX PL FL L 20&times;/0.40 CORR
-*** HCX PL FL L 63x/0.70 CORR
+*** HCX PL FL L 63&times;/0.70 CORR
-*** HCX PL APO 100x/1.47 OIL CORR&nbsp;TIRF</div></div>
+*** HCX PL APO 100&times;/1.47 OIL CORR&nbsp;TIRF</div></div>
-* confocal bio-imaging system: ''BD Pathway 435''
+* high-throughput microplate imager: ''Operetta CLS'' (PerkinElmer)
-* integrated system of Atomic Force Microscopy AFM, confocal microscopy and Raman spectroscopy (''NTEGRA Spectra, NT-MDT Co.'')
+* confocal bio-imaging system: ''BD Pathway 435'' (BD Biosciences)
-* inverted fluorescent microscope (''Leica DMI3000B'')
+* integrated system of atomic force microscopy (AFM), confocal microscopy and Raman spectroscopy (''NTEGRA Spectra'', NT-MDT Co.)
+* inverted fluorescent microscope (''DMI3000B'', Leica)
 * optical tweezers
@@ Line 109: / Line 111: @@
 *microfluidic platform for cell culturing ''CellASIC ONIX''
 *microPIV platform: ''Nikon Eclipse E-50i'' epi-fluorescent microscope with double-shutter CCD camera and double-pulsed Nd:YAG laser
-*stereo microscope
+*stereo microscopes
-*high precision syringe pumps (''neMESYS'', ''Cetoni GmbH'')
+*high-precision syringe pumps (''neMESYS'', Cetoni GmbH)
+*high-precision peristaltic pumps (''peRISYS'', Cetoni GmbH)
 *syringe pumps, peristaltic pump, micro gear pump
 *high-speed intensified CMOS camera (''Lambert HiCAM 500'')
-*high-speed CMOS camera (''pco.1200 hs, PCO IMAGING'')
+*high-speed camera (''Photron'', ''FASTCAM Mini AX100'' 540K-M-8GB)
-*double-shutter CCD camera (''SensiCam, PCO IMAGING'')
+*high-speed CMOS camera (''pco.1200 hs'', PCO IMAGING)
-*equipment for microfluidic chip fabrication by soft photolithography, i.a. spin coater, plasma cleaner, UV curing system, laboratory ovens, heating plates
+*double-shutter CCD camera (''SensiCam'', PCO IMAGING)
+*equipment for microfluidic chip fabrication by soft photolithography, ''i.a.'' spin coater, plasma cleaner, UV curing system, laboratory ovens, heating plates
 *equipment for microfluidic chip fabrication by micro-milling
+*3D printer (Zortrax ''Inventure'')
-===Nanofibres Electrospinning Unit===
+===Biomaterials Unit===
-* complete experimental setup for polymer nanofibres fabrication by electrospinning with necessary accessories for chemistry such as: fume hood, chemical balance, centrifuge, vortex, magnetic stirrer, water bath, refrigerated/heating circulators, etc.
+*setup for polymer nanofibers fabrication by electrospinning equipped with environmental chamber
+*3D bioprinter (Cellink, ''INKREDIBLE'' S-10002-001)
+*lyophilizator (LABCONCO, ''FreeZone 2.5 Liter -84C Benchtop Freeze Dryer'' 710201030)
+*plasma cleaner (Diener, ''ZEPTO PCCE'')
+*fume hood
+*accessories: analytical balances, centrifuges, vortexes, magnetic stirrers, blood roller mixer, water baths, refrigerated/heating circulators, conductometer, pH-meter, vacuum pump, pipettes, etc.
 ===Other equipment===
+*laser plotter equipped with Jasper X0-30 femtosecond laser (''Fluence'', Poland), designed for micromachining, cutting, and surface structuring of a wide range of materials (e.g., glass, plastics)
+*high-energy, double-pulsed Nd:YAG nanosecond laser (''EKSPLA NL303D'')
+*high frequency 1&nbsp;kHz pulsed Nd:YAG laser (''EKSPLA NL202'')
+*double-pulsed Nd:YAG laser (''SoloPIV'', New Wave Research Inc.)
 *solenoid micro-valves
 *pressure sensors
 *temperature sensors
-*high-energy, double-pulsed Nd:YAG nanosecond laser (''EKSPLA NL303D'')
-*high frequency 1&nbsp;kHz pulsed Nd:YAG laser (''EKSPLA NL202'')
-*double-pulsed Nd:YAG laser (''SoloPIV, New Wave Research Inc.'')
 *accessories like camera lenses, microscope objectives, mirrors, lenses, halogen lamps, sensors of temperature, pressure, humidity, viscosity and others
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