A µ-Slide with a porous glass membrane for transmigration and transport studies under both static and flow conditions

  • Brilliant optical quality through the thin, porous glass membrane
  • Full fluidic access to the apical and basal sides of adherent cells in numerous applications
  • Different pore sizes available for various cell types
  • Suitable for the establishment of lung models with air-liquid interface (ALI)

Applications

  • Trans-endothelial migration under flow conditions
  • Co-cultivation of cell layers and transport in 2D or in a 3D gel matrix
  • Apical-basal cell polarity assays
  • Skin and lung models with liquid-air interface
  • Cell barrier model assays with apical-basal gradients
  • Cell migration assays based on filters and porous membranes

Salvermoser, Melanie, et al. “Myosin 1f is specifically required for neutrophil migration in 3D environments during acute inflammation.” Blood, The Journal of the American Society of Hematology 131.17 (2018): 1887-1898. 10.1182/blood-2017-10-811851
Read article

Rohwedder, Ina, et al. “Src family kinase-mediated vesicle trafficking is critical for neutrophil basement membrane penetration.” Haematologica (2019). 10.3324/haematol.2019.225722
Read article

Transmigration Under Flow

Shown here is a time lapse movie of a transmigrating human HL-60 cell that was seeded into the lower channel of a µ-Slide Membrane ibiPore Flow (pore size 3 µm). Under flow conditions, the cell transmigrates in vitro through the porous glass membrane into the upper channel that was filled with a 3D collagen matrix containing the chemoattractant fMLP.

Live cell imaging of a transmigrating single SiR-Actin-stained cell (indicated by pseudo colors) by spinning disc confocal microscopy (20x objective) with a time interval of 30 s. The arrow indicates the direction of flow at a shear stress of 1 dyne/cm2; Scale bar = 10 um. Courtesy R. Pick, Munich, Germany.

Specifications

Total coating area4.50 cm²
Bottomibidi Polymer Coverslip

Main Channel (Lower Channel)

AccessLuer port, accessible with female Luers
Volume50 µl
Height0.4 mm
Length25 mm
Width5 mm
Growth area1.25 cm²

Upper Channel

AccessReservoir port, accessible with 20/200 µl pipet tips
Volume55 µl
Height over membrane1.3 mm

ibiPore Membrane

MaterialSiO2 (glass)
Thickness0.3 µm (300 nm)
Membrane size2 mm x 2 mm
Porous area1.77 mm x 1.84 mm
Restrictions for objective lensesWorking distance >0.5 mm
Pore layoutHexagonal spacing

Available Variations

Pore size
0.5 µm
3 µm
5 µm
8 µm
Porosity
20%
5%
5%
5%
Pore-to-pore distance
1 µm
12 µm
20 µm
32 µm

Which variation is suitable for my cell type?

Technical Features

  • SiMPore’s G-FLAT™ Microporous Glass Membranes
  • Cross-channel structure with a porous optical membrane in between
  • Excellent optical properties comparable to a glass coverslip
  • Pore sizes 0.5 µm, 3 µm, 5 µm, or 8 µm available
  • Membrane thickness 0.3 µm (300 nm)
  • For use with objective lenses with a working distance >0.5 mm
  • Fully compatible with the ibidi Pump System
  • Defined shear stress and shear rate levels
    (For details see Application Note 11)

The Principle of the µ-Slide Membrane ibiPore Flow

The μ-Slide Membrane ibiPore Flow consists of a horizontal porous glass membrane that is inserted between two channels. The upper channel is a static reservoir above the membrane. The lower channel is a perfusion channel for applying defined shear stress on cells, which are attached to the membrane. The upper and the lower channel communicate with each other only across the membrane.

Recommended Pore Sizes and Porosities for Different Applications

What Is the Porosity?

Porosity refers to the void volume fraction of the membrane. It is defined as the volume of the pores divided by the total volume of the membrane.

Membrane Pore Size and PorosityApplicationsExample Cell Types
0.5 µm pores,
high porosity (20%)
Permeability and transport studies,
co-culture models
Lung cells, epithelial cells
3.0 µm pores,
low porosity (5%)
Transendothelial migrationLeukocytes (e.g., neutrophils), lymphocytes (e.g., T cells or B cells)
5.0 µm pores,
low porosity (5%)
Invasion, migrationMonocytes, macrophages, lymphocytes (e.g., T cells or B cells)
8.0 µm pores,
low porosity (5%)
Invasion, migrationTumor and cancer cells, endothelial and epithelial cells, fibroblasts, osteoblasts, melanoma cells, glioma cells

The bigger the cell, the bigger the recommended pore size.

Intended Use

These applications have been tested by the ibidi R&D team or by our customers.

Endothelial Barrier Assays

A cell monolayer is cultivated on one side of the membrane.

Cells on the lower side of the membrane

Cells on the lower side of the membrane with defined shear stress

Cells on the upper side of the membrane

Co-Culture and Cell Barrier Assay

Two separate cell monolayers are cultivated on each side of the membrane. With this method, signaling, co-culture, and transport studies are possible.

Co-culture of two different cell types on both sides of the membrane

Apical-Basal Cell Polarity Assays

Chemical factors inside of a 3D gel matrix lead to the polarization of a cell monolayer that is cultured on the other side of the membrane.

Cells on the lower side of the membrane with a 3D gel matrix in the upper channel

Cells on the lower side of the membrane with a 3D gel matrix with embedded cells in the upper channel

Potential Use

The following examples illustrate further potential product uses. ibidi has not yet tested these applications in-house, therefore we cannot provide specific protocols. However, from a technical point of view, these applications should be possible.

We would appreciate your feedback for any application of the µ-Slide Membrane ibiPore Flow that works for you. Please send your images or videos with a short description to info@ibidi.de and we will be happy to publish them on our website to support the scientific community (e.g., as User Protocol).

Trans-Membrane Migration in 2D

A cell monolayer is cultivated on one side of the membrane to observe cellular trans-membrane migration.

Transmigration of suspended leukocytes

Rolling, adhesion, and transmigration of leukocytes, for example, under defined shear stress through a monolayer of cells on the lower side of the membrane

Cell Transport in a 3D Gel Matrix

Under flow conditions, the rolling, adhesion, and transmigration of leukocytes towards chemoattractant-producing cancer cells in a 3D matrix can be observed.

Transmigration of cells across a cell monolayer on the membrane into a 3D gel matrix with embedded cells

Non-Recommended Applications

Due to technical reasons, the following applications will not work with this product and should be avoided.
This product is not intended for:

  • Perfusion of the upper channel
  • Perfusion of both channels
  • Trans-membrane flow
  • Filter applications

Application Examples

Immunofluorescence Staining of Endothelial Cells

The µ-Slide Membrane ibidPore Flow is ideally suited for high-resolution microscopy of immunofluorescence stainings, due to the brilliant optical properties of the membrane.

Human endothelial cells on 3 µm ibiPore membrane after coating with Collagen Type IV according to Application Note 08 (PDF). Immunofluorescence overlay image of phase contrast, DAPI (blue), VE-cadherin (green), F-actin (red).

Phase Contrast Microscopy of Fibroblasts

Different cell types, such as fibroblasts, can be cultured and imaged on the thin, porous membrane of the µ-Slide Membrane ibiPore Flow.

Fibroblasts on 3 µm ibiPore membrane. Phase contrast image, objective lens 4x.

Lung Model With Air-Liquid Interface (ALI)

The dual channel system makes the µ-Slide Membrane ibiPore Flow ideal for the establishment of lung models with an air-liquid interface (ALI). A549 lung cells (adenocarcinomic human alveolar basal epithelial cells) were seeded on the upper side of the membrane and the lower channel was filled with culture medium. After cell adherence, the culture medium in the upper channel was replaced with air to establish an air-liquid interface. For imaging, the air was replaced with cell culture medium.

Phase contrast microscopy of A549 lung cells cultured on the µ-Slide Membrane ibiPore Flow on an ALI. Pore size 8 µm, 5% porosity. Data by Dr. Anita Reiser, Chair of Prof. Rädler, Ludwig-Maximilians-University Munich.

Cells on upper side of the membrane

Cells on lower side of the membrane

Phase contrast microscopy of A549 lung cells cultured on the µ-Slide Membrane ibiPore Flow, pore size 3 µm, 5% porosity. Left: cells were seeded on the upper side of the membrane. Right: cells were seeded on the lower side of the membrane. Data by Dr. Anita Reiser, Chair of Prof. Rädler, Ludwig-Maximilians-University Munich.

SEM Images of the Porous Glass Membrane


3 µm pores


3 µm single pore


0.5 µm pores


0.5 µm single pores

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