Organ-on-a-chip
Quick Start Guide v0.2

Single slot inserts

4-slot inserts

2

Top layer

Middle layer

Bottom layer

Chip structure3Organ on a chip – available inserts
Single slot OOC 45x15 with 0.4mm
middle layer:
Top insert 001.04.xx
bottom insert 003.07.xx

Single slot OOC 45x15 with 2.0mm
middle layer: Top insert 003.08.xx
Bottom insert 003.09

4-slot OOC 45x15 with 0.4mm
middle layer:
Top insert 003.10.xx
Bottom insert 003.11.xx

4-slot OOC 45x15 with 2.0mm
middle layer:Top insert 003.10.xx
Bottom insert 003.12.xx

4

Chip assembly – In Fluidic Connect Pro(recommended for single-slot inserts)

Note: tubing can be assembled before placing the chip(s) or afterwards. See
pages 11-15
Place the bottom
layer in the bottom
insert of the chipholder, with thegasket up.

Fill the gasket of thechip with water, until
it is completelyfilled.

Place the middle
layer with seeded
cells (for cellseeding, see p.9) on
the bottom layer ofthe chip in thechipholder, with themembrane down.

5Fill the middle layerof the chip withwater, untill thecavity is completelyfilled.

Place the top layerof the chip on the
middle layer in thechipholder, with thegasket down.

Close thechipholder. Fix theassembly, using thegripper.

6

Chip assembly – submerged(recommended for 4-slot inserts)

Place the bottom
layer of the chip in awater-filled petridish filled with thegasket on top.

Place the middle
layer with seeded
cells (for cellseeding, see p.9) on
the bottom layer ofthe chip in the petridish, with themembrane down.

Place the top layerof the chip on the
middle layer in the
petri-dish, with thegasket down.

7

Make sure the entire
chip is submerged.

Carfully pick thechip assembly fromthe dish.

Place the chipassembly in one of
the slots in thebottom insert of thechipholder and closethe chipholder.

8

Cell culture
Cell seeding
Cells can be seeded
directly by
pipetting the cellsuspension on the
middle layer.
Recommended
seeding densities
and membranecoatings are listed
in the table below.
Seeding densities
reported in thetable are meant forreaching confluentcell layers in 72hours.

Celltype

Seeding density[cells/cm 2]

Time forCoating
confluency
[h]

HUVEC

140-350 x103

72

Fibronectin 10-100

2

A549

35 x103

72

Fibronectin 10-1002

Calu-3300 x10372

No coating-

9Density
Time before[μg/cm 2] assembly[h]-Perfusion
Cells cultured on
the membrane aremechanicallystimulated by thefluid flow.
The different
height of thechannels results indifferent stress on
the cells (for thesame flow rates).

Shear stress τFlow rate q[dyne/cm2][µl/min]TOP

BOTTOM

0.01

47

80.05233

390.1467

780.52335

389

1-7785-3890

Top flow cell

The curves
representing theflow rate as afunction of shear
stress on themembrane in thetwo channels isreported in thisgraph.

Bottom Flow cell

4.0E+03

Flow rate [µl/min]

3.5E+03
3.0E+03
2.5E+03
2.0E+03
1.5E+03
1.0E+03
5.0E+02
0.0E+00
0123

Shear stress [dynes/cm²]

10An indication on
the flow rates
needed to achieve
specific shear
stresses can befound in this table.
Flow rates higher
than 4 ml/min arehighly notrecommended.

45

Tubing
Tube preparation

To prepare the tubing before use, use a tube cutter. Make sure a sharp blade isused and cut tubes according to the instructions of the tube cutter
Cut all tubes at the same length.

Tubing assembly – tubing and ferrule location
There are 6
connection holes inthe top inserts at
both ends of thechip. Only the holes(2, 5 and 8, 11) inthe middle at eachside need to beused

For assembly
information, see p.
12-15.

11

Tubing assembly – after closing the chip holder
Add ferrules in topinsert at thelocations of the inand outlet-channels.
Preferably use
ferrules with 1/16”
inner diameter foroutlet channels andferrules with an
inner diameter of1/32” for inlet
channels.

Close and fix thechipholder using thegripper. Make surethe ferrules stay inplace.

Press tubing tight in
the openings withferrules. Make surethe outer diameterof the tubing
correspondends with
the inner diameter.

12

Your setup is readyto use.

13

Tubing assembly – before closing the chipholder
Put tubing throughthe holes of the top
insert which wouldconnect to the inand outlet-channelsof the chip if thechipholder is closed.
Preferably use
tubing with 1/32”
outer diameter for
inlet channels andtubing with 1/16”
outer diameter foroutlet channels.
Slide ferrules over
the tubing (makesure the innerdiameter of theferrules correspondswith the outer
diameter of thetubing)Make sure thetubing sticks out of
the ferrules.

Press the tubing andferrules back into
the top insert. Make
sure the end of thetubing is on thesame level as theend of the ferrule.
The best way is to
first press thetapered part of theferrule in the insertopening, then press
the tubing in theferrule.

14

If the chip is not yet
in the holder, place
the chip in thechipholder.
Close and fix thechipholder using thegripper. Make surethe ferrules stay inplace.

Your setup is readyto use.

Note: After opening the chipholder, the top layer of the chip(s) may stick to theferrules. Please be cautious when opening the chipholder.

For more information go to: https://www.micronit.com/products/organ-on-a-chip.html

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Phone +31 53 850 6 850 Fax +31 83 850 6 851 E-mail info@micronit.com Internet www.micronit.com