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Cell Culture & Microscopy

Cell Culture & Microscopy

Cell Based Perfusion Assays

In vivo, several adherent cell types are exposed to mechanical shear stress in biofluidic systems like blood vessels. This mechanical stimulus has a great impact on the physiological behavior and adhesion properties of cells.

The µ-Slides were especially designed for conveniently performing flow assays. These consumables are tested for static and perfusion cell cultures. Being also compatible with any inverted microscopy technique they are ideally adapted to parallel perfusion assays directly on the microscope or inside the incubator.

Depending on the experimental requirements, peristaltic pumps, syringe pumps, or even simple gravity flow can be used. Click here for various pumping solutions or contact ibidi for personal consulting.


Experimental End Points Using µ-Slides

  • real-time monitoring of morphological changes in living cells
  • protein localization using fluorescence
  • profiling secreted proteins in conditioned media
  • cell adhesion
  • gene expression profiling after cell detachment

Adhesion Assay

Cell Culture & Microscopy
Example: blood cells to protein surfaces
Application: rolling and adhesion of suspended cells like platelets, leukocytes, monocytes on substrates like adhesion proteins or confluent cell monolayers
Flow characteristics: continuous laminar
Recommended pumps: syringe pumps, ibidi pump system, peristaltic pump
Duration: 20min - 2 hours
Recommended slides: µ-Slide VI, µ-Slide I Luer
Experimental environment: room temperature or incubation conditions*
* 37°C, 5% CO2 in air, 90% rel. humidity


Cell Culture Under Shear Stress

Cell Culture & Microscopy
Example: endothelial cells under flow
Application: influence of shear stress to endothelial cells, preparing cells while mimicking in vivo perfusion conditions, antibody stainings, formation of plaques on endothelium, biofilm formation of microorganisms
Flow characteristics: continuous laminar, non uniform, oscillating for turbulence simulation
Recommended pumps: ibidi pump system
Typical duration: 12 hours up to several weeks
Recommended slides: µ-Slide VI, µ-Slide I Luer,
µ-Slide y-shaped
Experimental environment: incubation conditions*
* 37°C, 5% CO2 in air, 90% rel. humidity


Stop Flow Experiments

Cell Culture & Microscopy
Example: Ca2+-imaging
Application: defined medium exchange for optimal feeding, online drug delivery, live stainings
Flow characteristics: laminar, short periods of flow inbetween non flow phases
Recommended pumps: manual liquid delivery (pipette, valve, and gravity flow, syringe etc.), syringe pumps, peristaltic pump
Typical duration: 12 hours up to several weeks
Recommended slides: µ-Slide VI, µ-Slide I Luer,
µ-Slide I


3D Cell Culture: Interstitial Flow

Cell Culture & Microscopy
Example: tumor cells in 3D gel matrix (e.g. collagen)
Application: 3D cultures of cells and tissue (hepatocytes, fibroblasts, muscle cells, kidney cells, stem cells)
Flow characteristics: continuous interstitial or stop flow interstitial
Recommended pumps: ibidi pump system
Typical duration: 12 hours up to several weeks
Recommended slides: µ-Slide VI, µ-Slide I Luer
Experimental environment: incubation conditions*
* 37°C, 5% CO2 in air, 90% rel. humidity


ECIS Flow Module

Cell Culture & MicroscopyThe ECIS technology allows measuring even small morphological changes with electrical signals, with a minimum of disturbance, and without any stainings. The measurements are automated and therefore highly reproducible with a minimum of work.


Shear Stress in General

Cell Culture & Microscopy... is the mechanical force induced by friction of liquid to the distal cell membrane. Cells might be able to countervail deformations caused by shear stress by rearranging their cytoskeleton. Other shear stress dependent effects are changes in metabolism, gene expression, and differentiation. Physiological shear stress values vary from 0.5dyne/cm² (small veins) to 20dyne/cm² (peaked flow in abdominal aorta).

Cell Culture & Microscopy
Turbulent Flow

... near surfaces is characterized by changes in flow rate and direction. For simulating turbulent flow it is accepted to use laminar flow of oscillating direction within half a second. This is because turbulent flow can not be achieved in microchannels at physiological flow regimes for physical reasons.

 

Laminar Shear Stress

Cell Culture & Microscopy... is expected in most healthy biological vessel like arteries and veins. Experimentally it is achieved by per-fusing medium through microchannels of low heights. The observation area of homogeneous laminar shear stress includes the whole channel area without small stripes close to the side walls and near the reservoirs (both in the range of the channel heights). For homo-geneous laminar shear stress experiments µ-Slide I Luer, µ-Slide VI, and µ-Slide I are recommended.


Non-Uniform Laminar Shear Stress

... occurs at branching sites and other obstacles of vessel in vivo. Experimentally it can be achieved by spatially varying flow rates. It can be used for investigating cells at different shear stresses in one sample. More advanced it can be used for studying cells and their communication at positions with strongly varying shear stress. The µ-Slide y-shaped was designed for studies of non-uniform shear stress. In the branched region the prevalent shear stress is approximately half of the regions with only the single channel. At the branching point experimentators should refer to our numerical simulations for exact shear stress values. Please contact us for personal consulting.