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Phase-Guide™

Caco-2 tubule

MediumCollagen gel

Well layout

To perfuse your cultures: place on the OrganoFlow® for programmable rocking.

Please check protocols and the rocker manual on our website for instructions.

ECM Channel Perfusion Channel Observation Window

Gel Inlet/Outlet (green)Used to access the collagen gel.

Top Perfusion Channel inlet and outlet (red), bottom Perfusion Channel inlet and outlet (orange) Inlets used to add medium, reagents or compounds to the Caco-2 tubules. (Apical or basal side)

The Observation Window (blue)Used for imaging your culture.This is where the three channelscome together and makecontact.

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OrganoPlate® Caco-2 in a nutshell Assay Ready Gut Tubules

1 2 3 4 . . .ABCD...

1 2 3 4 . . .ABCD...

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time (days)

TEER

(ohm

*cm

2)

800

600

400

200

0

Caco-2 over time

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20 hours exposure of healthy Caco-2 tubules to Staurosporine dilutions in the OrganoPlate®

Nor

mal

ized

TEE

R

0:02 0:05 0:11 0:22 0:45 1:30 3:00 6:00 12:00 0:00

1.2

1

0.8

0.6

0.4

0.2

0

0uM9,7nM39nM156nM625nM2,5uM10uMnegative

Experiments in the OrganoPlate® Caco-2

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Culture readout examples

The four steps to get started with the OrganoPlate® Caco-2

Applications

CellsECM-gelNumber of tissue chipsNumber of control chipsCompound access to tissue tubulesMicro!uidic lane widths Internal volumesGel-medium interface surfacePlate format

Materials

Micro!uidic lane height PhaseGuide™ dimensions Medium volumePerfusion

Compatible assays

Readouts

Drug-induced toxicity, Intestinal Permeability, DDI studies with P-gp transporter Caco-2 cell line, Human Caucasian colon adenocarcinoma, Wild-TypeCollagen 1 (Rat Tail)382Apical & basal (in- and outside)

Top and bottom perfusion lane: 320 μm. Middle lane: 360 μmTop and bottom perfusion lane: 1.1 μL. Middle lane: 0.87 μL0.5 mm2

SBS-standard 384-well plate, 127.76 x 85.48 x 14.8 mm, (l x w x h) (17.3 mm height with lid)Top plate: virgin polystyrene. Plate bottom: optical quality 170 μm glass (1.5H coverslip thickness). Micro"uidics: glass, proprietary polymers, biocompatible and low compound-absorbing.220 μm100 x 55 μm (w x h)15-75 μL in each wellGravity driven, pump free, typical shear forces in tubule 0 - 3 dyne/cm2

• Immunostaining • DNA isolation • RNA isolation• Cell viability assays• Transport assays

Imaging (phase contrast, wide#eld "uorescence, confocal), plate reader (absorption, "uorescence, luminescence), sampling (ELISA, PCR, sequencing, MS, biochemistry)

• Migration assays• Barrier integrity assay• Western blot• Mass spectometry• Adhesion assays

Product speci"cations

• ELISA• TEER

Unpack the OrganoPlate® Caco -2 and remove shipment medium accordingto protocol

Incubate and perfuse the OrganoPlate® Caco-2 overnight

Perform your experiment

Check for the latestprotocols:

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